Reviews of

Environmental Contamination
and Toxicology
VOLUME 193
 Reviews of
Environmental Contamination
and Toxicology
Editor
David M. Whitacre

Editorial Board
Lilia A. Albert, Xalapa, Veracruz, Mexico · Charles P. Gerba, Tucson, Arizona, USA
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George W. Ware, Tucson, Arizona, USA

Founding Editor
Francis A. Gunther

VOLUME 193
 Coordinating Board of Editors

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Reviews of Environmental Contamination and Toxicology

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 Foreword

International concern in scientifific, industrial, and governmental communi-

ties over traces of xenobiotics in foods and in both abiotic and biotic envi-
ronments has justified
fi the present triumvirate of specialized publications in
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our food, our feeds, our homes, recreational and working surroundings, our
domestic animals, our wildlife and ourselves. Tremendous efforts world-
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and toxicology of the chemicals loosed upon the earth. Among the sequelae
of this broad new emphasis is an undeniable need for an articulated set of
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literature produced by these emerging areas of science together with docu-
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consequences.
Bulletin of Environmental Contamination and Toxicology (Vol. 1 in 1966)
for rapid publication of short reports of significant
fi advances and

v
vi Foreword

discoveries in the fi
fields of air, soil, water, and food contamination and
pollution as well as methodology and other disciplines concerned
with the introduction, presence, and effects of toxicants in the total
environment.
Archives of Environmental Contamination and Toxicology (Vol. 1 in 1973)
for important complete articles emphasizing and describing original
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aspects of chemical contaminants in the environment.
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Coordinating Board of Editors

 Preface

The role of Reviews is to publish detailed scientificfi review articles on all

aspects of environmental contamination and associated toxicological con-
sequences. Such articles facilitate the often-complex task of accessing and
interpreting cogent scientific
fi data within the confifines of one or more closely
related research fields.
In the nearly 50 years since Reviews of Environmental Contamination
and Toxicology (formerly Residue Reviews) was fi first published, the number,
scope and complexity of environmental pollution incidents have grown
unabated. During this entire period, the emphasis has been on publishing
articles that address the presence and toxicity of environmental contami-
nants. New research is published each year on a myriad of environmental
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covery and reporting of new environmental contamination cases, creates
an increasingly important function for Reviews.
The staggering volume of scientific
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and global warming. Unfortunately, these very worrisome issues are now
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challenges.
The ultimate role of publishing scientific
fi research is to enhance under-
standing of the environment in ways that allow the public to be better
informed. The term “informed public” as used by Thomas Jefferson in the

vii
viii Preface

age of enlightenment conveyed the thought of soundness and good judg-

ment. In the modern sense, being “well informed” has the narrower meaning
of having access to suffi ficient information. Because the public still gets most
of its information on science and technology from TV news and reports, the
role for scientists as interpreters and brokers of scientificfi information to
the public will grow rather than diminish.
Environmentalism is the newest global political force, resulting in the
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sive confrontation? These matters are of genuine concern to governmental
agencies and legislative bodies around the world.
For those who make the decisions about how our planet is managed,
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trialized culture is a dynamic challenge, for the old, established materials
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able to federal and state regulatory agencies, public health officials,
fi and
environmentalists.
Reviews publishes synoptic articles designed to treat the presence,
fate, and, if possible, the safety of xenobiotics in any segment of the envi-
ronment. These reviews can either be general or specifi fic, but properly lie
in the domains of analytical chemistry and its methodology, biochemistry,
human and animal medicine, legislation, pharmacology, physiology, toxicol-
ogy and regulation. Certain affairs in food technology concerned spe-
cifi
fically with pesticide and other food-additive problems may also be
appropriate.
Because manuscripts are published in the order in which they are
received in final form, it may seem that some important aspects have been
neglected at times. However, these apparent omissions are recognized,
and pertinent manuscripts are likely in preparation or planned. The fi field is
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the Editorial Board earnestly solicit authors and suggestions of under-
represented topics to make this international book series yet more useful
and worthwhile.
Justification
fi for the preparation of any review for this book series is that
it deals with some aspect of the many real problems arising from the pres-
ence of foreign chemicals in our surroundings. Thus, manuscripts may
encompass case studies from any country. Food additives, including pesti-
cides, or their metabolites that may persist into human food and animal
feeds are within this scope. Additionally, chemical contamination in any
manner of air, water, soil, or plant or animal life is within these objectives
and their purview.
 Preface ix

Manuscripts are often contributed by invitation. However, nominations

for new topics or topics in areas that are rapidly advancing are welcome.
Preliminary communication with the Editor is recommended before volun-
teered review manuscripts are submitted.

Summerfield,
fi North Carolina D.M.W.
 Table of Contents

Foreword ..................................................................................................... v
Preface ......................................................................................................... vii

Remediation Technologies for Organochlorine-Contaminated Sites

in Developing Countries ............................................................................ 1
Alberto Bezama, Rodrigo Navia, Gonzalo Mendoza, and
Ricardo Barra

Chemistry and Fate of Triazolopyrimidine

Sulfonamide Herbicides ............................................................................. 31
Thomas W. Jabusch and Ronald S. Tjeerdema

Parameters for Carbamate Pesticide QSAR and PBPK/PD Models

for Human Risk Assessment ..................................................................... 53
James B. Knaak, Curt C. Dary, Miles S. Okino,
Fred W. Power, Xiaofei Zhang, Carol B. Thompson,
R. Tornero-Velez, and Jerry N. Blancato

Persistent Organic Pollutants in Vietnam: Environmental

Contamination and Human Exposure ..................................................... 213
Tu Binh Minh, Hisato Iwata, Shin Takahashi,
Pham Hung Viet, Bui Cach Tuyen, and Shinsuke Tanabe

Index ............................................................................................................. 291

xi
Rev Environ Contam Toxicol 193:1–29 © Springer 2008

Remediation Technologies for Organochlorine-

Contaminated Sites in Developing Countries

Alberto Bezama, Rodrigo Navia, Gonzalo Mendoza,

and Ricardo Barra

Contents
I. Introduction ............................................................................................................ 1
II. Chlorinated Hydrocarbons as Key Environmental Pollutants........................ 4
III. Implementing Remediation Technologies for Organochlorine-
Contaminated Sites................................................................................................ 8
A. Soil.................................................................................................................... 11
B. Groundwater ................................................................................................... 13
IV. Definitions
fi of An Integrated Approach ........................................................... 16
A. Soil Washing ................................................................................................... 17
B. Wastewater Treatment .................................................................................. 17
C. Characterization of Waste Streams ............................................................. 19
D Windrow Composting .................................................................................... 20
V. Discussion ............................................................................................................. 20
Summary ............................................................................................................... 21
Acknowledgments ............................................................................................... 22
References ............................................................................................................ 22

I. Introduction
Despite its importance in human life, until recently the relationship between
soils and human health has been undervalued, especially in least developed
countries. Currently, a holistic approach has been incorporated to identify
best practices in soil science, defi
fining it as “the task of all people concerned
with the soil to direct their interest, not just towards the physical, chemical,
and biological aspects, but also to those environmental, economic, social,
legal, and technical aspects that affect soil use” (Abrahams 2002; Fent
2003). Considering this defi finition, the European Union (EU) as well as
most developed countries have recognized organochlorine-contaminated

Communicated by Lilia Albert

A. Bezama ( ), G. Mendoza, R. Barra
Environmental Sciences Center EULA—Chile, University of Concepción, Barrio Universi-
tario s/n, Concepción, Chile.
R. Navia
Department of Chemical Engineering, University of La Frontera, Av. Francisso Salazar 01145,
Temuco, Chile.

1
2 A. Bezama et al.

sites as potential threats to the human health, threats that take different
forms, such as their inflfluence on water (e.g., drinking water resources), soil,
and air as well as their interrelationships, which can directly affect human
health (EC 2002; EP 2002; Bezama et al. 2004). Moreover, economic expan-
sion and industrial growth are linked with growing lack of “greenfields”
fi (a
term that defi fines all areas without previous history of development): the
supply of new building sites is limited and must contend with other compet-
ing uses, such as housing, recreation, nature, traffic, fi or agriculture (De
Sousa 2001; Tedd et al. 2001). Thus, cleaning and reusing contaminated
sites can be a meaningful alternative to address this issue, because most
contaminated sites are located in metropolitan centres and are, therefore,
prime candidates for urban development (Lorber et al. 2004).
The identification
fi of sites that pose a potential risk to human health and
ecosystems, the verifi fication of their actual pollution level, and assessment
of the involved risks are the first steps when managing a contaminated land
(Bezama et al. 2007a). Occurrence and distribution of soil problems in
Europe are infl fluenced by the diversity, distribution, and specifi fic vulner-
ability of soils across the continent, coupled with physical aspects such as
geology, relief, and climate (EEA 1999). About 550,000 sites across the EU
have been identifi fied as defi finitively or potentially contaminated, and the
best estimate is that 1.3 million contaminated areas will be registered,
although there is still a lack of information about the type and size of these
contaminated areas. An estimated overview of the contaminated sites situ-
ation in Europe can be observed in Fig. 1 (Bezama 2006).
Organochlorine compounds represent an important fraction of the pol-
lutants present in the identifiedfi contaminated sites in Europe, especially
considering those originating from industrial and agricultural activities,
which together are approximately 75% of the identifi fied potentially con-
taminated sites (EEA 1999, 2003). For example, from the approximately
70,000 sites suspected to be contaminated in Austria, 33,549 have been
registered as potentially industrial contaminated sites. Of these registered
sites, 163 have already been investigated, evaluated, and classified.
fi The data
show that ∼32% of the registered contaminated sites were contaminated
with organochlorine compounds, whereas in 2002 these were found as main
pollutants in ∼29% of the 163 evaluated sites (FEA 2002).
As in most South American countries, the magnitude of the Chilean
contaminated site problem has yet to be established. To date, two studies
have been conducted for the identifi fication and preliminary risk assessment
of sites under suspicion of contamination, considering the associated human
health and environmental risks (Fundación Chile 2004; Bezama et al.
2007a). The latter work is a case study in an industrial Region in South
Central Chile that is the second most important nationally in social and
economic terms. This preliminary investigation, based on historical data-
bases of industrial activities and without a more accurate identificationfi
process due to limited funding, identified fi nearly 510 sites as suspect of
 Remediation of Contaminated Sites 3

Fig. 1. Summary of the identifi

fied contaminated sites across Europe (Bezama 2006)
(Adapted from EEA 1999; EEA 2003; FEA 2002; Eurostat et al. 1995, Andersen
2000.)

contamination in the fifirst stage, from which approximately 10% were evalu-
ated as dangerous to human health. As expected, a large number of these
sites have an industrial origin. According to Seguel (2002), the total amount
of industrial wastes generated in this Chilean Region amounts to about
350,000 t/yr. Most of the generated waste corresponds to the forestry sector
(including sawmills, pulp and paper, cardboard, chipboards, and other com-
panies, comprising ∼21% of the regional waste generation) and to the iron
and steel industry (17%). Considering these industrial activities, it is possi-
ble that chlorinated compounds are pollutants in this Region’s contami-
nated sites.
The objective of this study is to define
fi the technical considerations to
be used when dealing with chlorinated-compound-contaminated sites,
especially focusing on sites contaminated with chlorinated hydrocarbons
(CHCs), chlorophenols (CPs), and polychlorinated biphenyls (PCBs), con-
sidering their occurrence in a developing economy such as Chile. These
defifinitions are then followed by the determination of an integral site reme-
diation technology that is based on the European experience but which
could be easily transferable to the reality of developing countries in South
America, specifically
fi considering the Chilean case.
4 A. Bezama et al.

II. Chlorinated Hydrocarbons as Key Environmental Pollutants

Chlorinated hydrocarbons have been found in old contaminated sites of

solvent, cleaning, and electrical industries as well as in sites of textile,
tanning, and pulp and paper industries as a result of their use as bleaching
agents. In this review, we discuss the most common and important
subgroups among chlorinated compounds, namely the CHCs, CPs, and
PCBs.
CHCs have been found in old wood storage sites, where they were used
as preserving agents. Trichloroethylene (TCE), perchloroethylene (PCE),
dichloromethane (DCM), 1,1,1-trichloroethane (TCA), or chloroform
(CFM) are used extensively in many industries (e.g., in metal degreasing,
dry cleaning, paint industries, organic synthesis), and as a result of their
common use, they can be a signifi ficant source of pollution in the atmos-
phere, soils, and aquifers (Jendrzejewski et al. 2001; Michael et al. 1991;
McCulloch and Midgley 1996; Stiber et al. 1998; Sturchio et al. 1998). Owing
to their toxicity, they can threaten both public health and sustainable biore-
source use and have actually generated worldwide concern as toxic envi-
ronmental contaminants (Bidelman and Olney 1974; Tanabe et al. 1994;
Guzzella et al. 2005; de Mora et al. 2004).
The principal industrial uses of CPs are in wood, textile, and leather
preservation. An additional important source for the introduction of these
compounds into the environment has been their application as pesticides
and the degradation products of common pesticides. Moreover, CPs are
by-products in the chloro-bleaching process of pulp and paper mills (Ohlen-
busch et al. 2000), an industry of great relevance to Chile; while world pulp
and paper production is estimated as 300 million t/yr, the Chilean produc-
tion reaches about 4.4 million t/yr (INFOR 2006), a number expected to
increase in the future because of the installation of several large pulp and
paper mills in South-Central Chile.
The Cluster Rule (EPA 1998) introduced in the United States regulates
the level of the 12 chlorophenols presented in Fig. 2 (McKague and Taylor
2001). The most important chlorophenols present in groundwater streams
in the U.S. and Germany are 2,4,5-trichlorophenol, 2,4,6-trichlorophenol,
2,4-dichlorophenol, 2-chlorophenol, 3-chlorophenol, and pentachlorophe-
nol (Kerndorff 1996). Moreover, these 6 compounds are on the list of the
most common contaminants present in groundwater streams at contami-
nated sites. Finally, although 2,4,5-trichlorophenol, 2,4,6-trichlorophenol,
2,4-dichlorophenol, 2-chlorophenol, and pentachlorophenol are in the third
group of priority contaminants according to EPA (1998), 3-chlorophenol
and 2,6-dichlorophenol could join the first or the second group as their
toxicity is not yet well established (Kerndorff 1996).
Another important group of organochlorine compounds are the PCBs.
Synthesis of these compounds was first fi described in 1881, and their com-
mercial production began at the end of 1920 (UNEP 2002), specifically fi
 Remediation of Contaminated Sites 5

2,4,5-Trichlorophenol 2,4,6-Trichlorophenol 2,3,4,6 Tetrachlorophenol

OH O H O H

Cl Cl Cl Cl Cl

Cl Cl

Cl Cl Cl

Pentachlorophenol 3,4,5-Trichlorocatechol 3,4,6- Trichlorocatechol

O H O H O H

Cl Cl O H Cl O H

Cl Cl Cl Cl Cl

Cl C l Cl

Tetrachlorocatechol 3,4,5-Trichloroguaiacol 3,4,6-Trichloroguaiacol

O H OH OH

Cl O H OCH 3 Cl OCH 3

Cl Cl Cl Cl Cl

Cl Cl Cl

4,5,6-Trichloroguaiacol Tetrachloroguaiacol Trichlorosyringol

OH OH OH

Cl OCH 3 Cl OCH 3 C H 3O OCH3

Cl Cl Cl Cl Cl

Cl Cl Cl

Fig. 2. Chlorophenols regulated by the Cluster Rule.

between 1929 and 1939. One of the most important characteristics of the
PCBs is the wide variability in their physicochemical properties (vapor
pressure, water solubility, and partition coefficients)
fi as a consequence of
their great number of congeners (209), varying from very low molecular
weight/high volatility compounds (such as monochlorobiphenyl) to not very
volatile/high molecular weight compounds (such as decachlorobiphenyl),
which determines their behavior and mobility in different environmental
compartments (Table 1). Such characteristics have resulted in international
standards to regulate use of these compounds in all their life-cycle stages
and in all their applications, e.g., recently in the European food industry
(EC 2006).
Total PCB global production has been estimated to reach 1.3 million ts
(Breivik et al. 2002). In Chile, as in other developing countries, PCBs have
been used for more than 30 yr in many different industrial applications
(Barra et al. 2004), principally as dielectric fl
fluid, whose use was prohibited
in 1982, in new transformers and condensers. However, their use in old
devices and later storage may present a high environmental risk. In July
 6

Table 1. Properties for polychlorinated biphenyls (PCB) congeners.

Name Number of IUPAC Molecular Vapor pressure Solubility in Number of

Formula (chloro-biphenyl) isomers numbers weight Log Kow % Cl (Pa) water (g/m3) identifi
fied isomersa

C12H9Cl Mono 3 1–3 188.7 4.3–4.6 18.79 0.9–2.5 1.21–5.5 3

C12H8Cl2 Di 12 4–5 233.1 4.9–5.3 31.77 0.008–0.6 0.06–2.0 12
C12H7Cl3 Tri 24 16–39 257.5 5.5–5.9 41.3 0.003–0.22 0.015–0.4 23
C12H6Cl4 Tetra 42 40–81 291.9 5.6–6.5 48.65 0.002 0.0043–0.01 41
C12H5Cl5 Penta 46 82–127 326.4 6.2–6.5 54.3 0.0023–0.051 0.004–0.02 39
C12H4Cl6 Hexa 42 128–169 360.8 6.7–7.3 58.9 0.0007–0.012 0.004–0.0007 31
Hepta 24 170–193 395.3 6.7–7 62.7 0.00025 0.000045–0.00001 18
A. Bezama et al.

C12H3Cl7
C12H2Cl8 Octa 12 194–205 429.8 7.1 66.0 0.0006 0.0002–0.0003 11
C12HCl9 Nona 3 206–208 464.2 7.2–8.16 68.7 — 0.00018–0.00012 3
C12Cl10 Deca 1 209 498.6 8.26 71.1 0.00003 1 × 10−6–1 × 10−7 1
a
Refers to the number of isomers identified
fi in commercial mixtures.
Source: Guitart et al. (1995); de Voogd et al. (1990); Ritter et al. (2005).
 Remediation of Contaminated Sites 7

Table 2. PCB amounts for Chilean regions.

Amounts of PCB in the

Region (L)

Region In use Stored Total (L)

I 699 22 721
II 62,014 156,408 218,442
III 10,767 56,267 67,034
IV 7,745 681 8,426
V 12,132 250 12,382
R.M. 77,235 5,005 82,240
VI 44,944 742 45,686
VII 173 173 173
VIII 80,645 20,378 101,023
IX 450 0 450
X 60 0 60
XI 0 0 0
XII 30,314 2,616 32,930
Total 327,005 242,542 569,547

Source: CONAMA (2005b).

2004, Chile ratifi

fied the Stockholm Convention, which established different
measurements to reduce and to eliminate POPs, including the prohibition
of use and reduction of by-product liberation. This agreement has served
as a diagnostic and assessment mechanism for these compounds at the
national level in Chile. The Chilean PCB Register, carried out in 2004,
which considered only dielectric fl fluids for condensers and transformers,
shows the following inventories for Chile in each region, both for current
use as well as for stored quantities (Table 2).
In the area of air quality, Barra et al. (2005) report that few PCB meas-
urements have been performed in Chile. Recent reports for Santiago de
Chile indicated values from 1.04 to 1.75 ng/m3, which makes it comparable
to other urban areas of the world. PCB levels in air particles smaller than
2.5 μm diameter (PM2.5) in Temuco, Chile, ranged from 0.67 to 1.7 μg/m3,
while in Santiago the levels ranged from 1.15 to 2.7 μg/m3. Unexpectedly,
differences observed between these two cities were not high, in spite of
differences in population density, indicating that the atmosphere in these
two Chilean cities is fairly well polluted with PCBs, even when compared
with other urbanized areas of the world (Mandalakis and Stephanou
2002).
Furthermore, there are very few offi ficially recognized contaminated sites,
and these are located mainly in heavily populated industrial areas as
Santiago and Concepción, Chile. However, these offi ficial numbers grossly
underestimate the real situation because of the existence of illegal and/or
8 A. Bezama et al.

Table 3. Distribution of the average sum of PCB concentrations in mussels through-

out different Chilean regions.

Average sum of PCBs

Geographic distribution Number of sampling points concentrations (ng/g d.w.)

North 4 25.3
North Center 3 37.2
South Center 3 12.4
South 3 15.3
South End 3 198.5

Source: Mendoza et al. (2006).

nonreported contaminated sites throughout the entire country (Barra et al.

2006). Available data are scarce and fragmentary; however, it is clear that
PCB contamination is widespread through the country, as even the concen-
trations reported for background levels are lower than in the Northern
Hemisphere (Barra et al. 2006; Borghini et al. 2005). PCB values in sedi-
ments from a highland lake have been reported, with quite low levels,
reaching maximum values of 2 ng/g dw, but with trends indicating an
increase in recent times (Barra et al. 2004). The most recent experience
(Mendoza et al. 2006) consisted of the spatial distribution analysis of PCBs
in mussels for the length of the Chilean coast. In this study, statistical analy-
sis of congeneric PCB composition indicated five groups according to their
molecular weight (number of chlorines), where the lighter congeners were
observed in areas corresponding to high latitudes with total PCB values of
298 ng/g dry weight (Table 3).

III. Implementing Remediation Technologies for

Organochlorine-Contaminated Sites
Remediation can be defined
fi as the improvement of a contaminated site, to
prevent, minimize, or mitigate damage to human health or the environ-
ment. It involves the development and application of a planned approach
that reduces the availability of contaminants to receptors of concern (CSM
1997). The selection of an appropriate technology is therefore a crucial
activity in a remedial project, where technical, economic, social, legal, and
environmental aspects must be considered and evaluated (DEPA 2002;
Bezama 2006). In general, it is possible to define fi two different types of
remediation processes: in situ and ex situ. In situ processes are carried out
in the same place where the pollutant is present, while ex situ processes are
carried out in on-site or off-site plants or facilities. On-site refers to the
place directly at the contaminated site, whereas off-site indicates that the
place of a remediation facility is located outside the contaminated site
(Hamby 1996; Guerin 1999; Bezama et al. 2004).
 Remediation of Contaminated Sites 9

Any conventional in situ remediation scheme, such as bioremediation or

soil vapor extraction, can be applied to the excavated soil, and because soil
is usually handled in a closed system, process operational control is greatly
simplifi fied. Aggressive remediation schemes, such as surfactant or acid
leaching of soil, can be applied without posing a threat to the environment.
Often this cannot be done in situ for fear that mobilized contaminants will
escape containment or will contaminate previously uncontaminated soil.
Additionally, the treatment process can be more easily monitored, and
unsuccessful processes may be abandoned with little environmental risk
because the pile remains encapsulated until satisfactory results are attained.
Furthermore, gradual, long-term remediation techniques can be used to
provide low-cost, low-risk remediation that would not be feasible in situ
(Lante 1991). This approach is similar to using in situ “natural attenuation”
(Lin and Puls 2003; Kao et al. 2001; Youcai et al. 2002) but allows for
greater mass transport control while the process continues.
In ex situ soil remediation, because wastes are removed from their origi-
nal locations, much of the original site can be quickly recovered. This
method can be a convincing advantage for redeveloping the affected areas
where a small portion of the site can be dedicated to gradual remediation
while the remainder of the site is redeveloped, or where one small site can
host the remediation for several similar locations (Mesania and Jennings
2000). On the other hand, in contrast to ex situ remediation, in situ tech-
nologies can be used at a site with little disruption to ongoing operations.
In situ treatments require neither heavy equipment for excavation nor large
aboveground surface areas for treatment technology equipment facilities.
Because in situ treatment does not involve movement of contaminated
materials in the location, it minimizes both human and environmental
exposure to contaminants. In contrast, transferring subsurface contami-
nated materials to the surface increases exposure risk for the same recep-
tors (Volkwein et al. 1999).
In addition, in comparison with ex situ technologies, in situ technologies
may be a more cost-effective and less intrusive means of remediating con-
taminated soils, sediments, and groundwater (Dupont 1993; Lin et al. 1996).
Excavating contaminated material as well as operating and maintaining
facilities for ex situ treatments often result in higher costs for treatment on
the surface. On the other hand, to use an ex situ approach, one must remove
contaminated soil from its original location and treat it above ground
(Mesania and Jennings 2000; Van Deuren et al. 2002; Bezama et al.
2007b).
To determine the remediation processes that could be useful for
organochlorine-contaminated sites, the physicochemical properties of some
relevant chlorinated compounds present in the soil–aquifer environment
need to be examined. In this case, compounds that are considered to be
representative for this analysis because of their toxicity and ubiquitous
presence are PCP, 2,4,6-trichlorophenol, and 2-chlorophenol (Table 4).
10 A. Bezama et al.

Table 4. Physicochemical properties of some chlorophenols.

Water
solubility* Specific
fi Boiling point °C Vapor pressure
Contaminant Kow (mg/L) gravity (760 mmHg) pK Ka (mmHg)

PCP 1.0 * 105 1.4 * 101 1.9278 311.0 4.7 0.0002

TCP 7.4 * 101 8.0 * 102 1.49 244.5 6.2 400a
CP 1.5 * 101 2.9 * 104 1.241 174.5 8.5 1b

PCP, pentachlorophenol; TCP, 2,4,6-trichlorophenol; CP, 2-chlorophenol.

a
At 12.1°C.
b
At 25°C.
*106 = infi
finite solubility.
Source: Adapted from Shiu et al. (1994).

PCP properties are of special importance for Chile because the intensive
forestry activity in the South Central region results in a large number of
sawmills and wood-related enterprises, where the Potentially Contami-
nated Sites are highly probable from PCP use, especially in those compa-
nies operating before 1999, when the Chilean government banned PCP use.
Until that date, more than 500 t PCP was imported and used for wood
impregnation, where sodium salt is the most common form used.
In Table 4, it can be observed that nonionic PCP has low solubility and
very low vapor pressure, which means, looking at its Kow value, that PCP
has a great affinity
fi for organic matter and will probably remain adsorbed
onto the organic part of the soil in the unsaturated zone. It is possible to
assume that nonionic PCP has a low Henry’s law constant, so that it will be
very difficult
fi to find PCP in the vapor phase. Regarding nonionic TCP, this
compound would be preferentially found in the vapor phase of the vadose
zone because of its high vapor pressure, moderated water solubility (Table
4), and high Henry’s law constant. The Kow for TCP is small, and TCP can
be assumed to adsorb lightly on organic matter of the soil. Finally, it is
possible to assume that nonionic CP would be preferentially dissolved in
the aqueous phase of the vadose zone because of its high solubility value,
low vapor pressure (see Table 3), and moderated Henry’s law constant. The
Kow value for CP is also small, and it is possible that this pollutant could
lightly adsorb onto soil organic matter. For the unsaturated zone, the situ-
ation would be as described in Fig. 3a, assuming as well an unsaturated zone
pH smaller than pKa and that all compounds are present in their nonionic
form.
All these considerations correspond to the pollutants in their original
form, not dissociated, although they may change depending on the pH of
the water and soil system, affecting each compound’s dissociation capacity
in dependence with its pKa value. These dissociated species may present
different behaviors in the surrounding environment and will have different
 Remediation of Contaminated Sites 11

(a) (b)

Dissolved CP Gaseous CP Dissolved Gaseous

Adsorbed PCP nonionic CP nonionic CP Dissolved
anionic PCP

Soil pore
p
Soil pore
p
Gaseous TCP Gaseous
nonionic TCP
Water pore
Water pore

Dissolved
Dissolved TCP
anionic TCP

Fig. 3. Possible contaminants (chlorophenols) distribution in the vadose zone:

(a) pH < pΚa and (b) pH = 7.

solubilities, Henry’s law constants, and vapor pressures. Thus, soil and
water pH would become an important factor when choosing or designing
a remediation process. If we consider that pH controls the presence of the
dissociated species in a soil–water environment, the dissociation curve for
each pollutant and its dependence with pH needs to be known. Taking as
example a pH value of 7 in the soil/water system, PCP would be preferen-
tially in the anionic form and its solubility will increase from 1.4 × 101
(nonionic PCP) to 104 mg/L (anionic PC-phenolate). In contrast, the situa-
tion for TCP and CP should be quite different, because at pH 7 TCP would
be present at about 50% in the anionic form and 50% in the nonionic form
and CP should be mainly present in the nonionic form in the aqueous phase.
Therefore, there might be only small quantities of nonionic TCP and CP
that remain adsorbed onto the soil pores (Fig. 3b).

A. Soil
A widely used remediation practice for soils polluted with hydrocarbons is
bioremediation. In recent years, bioremediation has been increasingly
selected to treat organochlorine compounds, because it is more publicly
acceptable as it relies on natural processes to treat contaminants (EPA
2000; GeoSyntec Consultants 2005). Bioremediation utilizes microorgan-
isms to degrade organic contaminants in soil, sludge, and solids, either
excavated or in situ. Microorganisms break down contaminants by using
them as a food source or cometabolizing them with a food source. Aerobic
processes require an oxygen source, and the end products typically are
carbon dioxide and water, although metabolites can also be found as inter-
mediates (Zink and Lorber 1995). In ex situ processes, optimal aerobic
12 A. Bezama et al.

conditions can be achieved adding structure materials (e.g., perlite) to an

optimal surface for bacterial growth. In these processes, the soil is arranged
in a pile and aeration is normally achieved using a vacuum pump. The
contaminated air passes through a biofi filter and is returned to the pile. In
addition, the percolated water is collected, cleaned in an aerated treatment
plant, and recirculated with nutrients into the pile to maintain its optimal
moistore content (Eiermann and Bolliger 1996).
One of the more often used soil bioremediation technologies is in situ
bioventing (Stehmeier et al. 1999), especially when dealing with CHC-
contaminated soils (Vogel 1996; Whyte et al. 2001; Gogoi et al. 2003). This
technology relies on an increase in the flow of air through the vadose zone
that provides oxygen in the subsurface to optimize natural aerobic biodeg-
radation, which becomes the dominant remedial process (Eiermann and
Bolliger 1996; Warith et al. 1999). It is known from the literature that
aerobic biological remediation processes are highly dependent on pollutant
bioavailability, oxygen, and nutrients (nitrogen and phosphorus). As a
result, in the ideal bioventing condition, the pollutants, oxygen, and nutri-
ents are all dissolved in the water phase, which is very difficult
fi to achieve.
Therefore, the main problem for biological in situ treatment is that usually
long process times are required (years), the microorganisms must be very
specific,
fi and the products of the biochemical reactions involved should be
determined to be innocuous. Another application is the soil–air extraction
in situ process, which relies on extraction of the air in the vadose zone
through a vacuum pump. This process is suitable for high vapor pressure
(>1500 Pa) contaminants in the soil matrix. The outlet air stream should be
further treated in a biofi
filter or an adsorption system (Hamby 1996; Bezama
2006).
Another interesting technological option that has gained much attention
recently is soil washing (Semer and Reddy 1996; DEPA 2002; Haapea and
Tuhkanen 2006). Soil washing is an ex situ technology that relies on water
as the fluid medium and energy to separate pollutants in nonsaturated
media (Mann 1999; Norris et al. 1999). Soil washing liberates pollutants
from the soil pores with mechanical energy, creating a pollutant suspension
or an emulsion in the wash fl fluid (Tokunaga and Hakuta 2002; Urum et al.
2005). An ultimate pollutant separation is required to recover a concen-
trated pollutant and a clean wash fluid. Pure water is normally used, but
some alternatives include addition of organic solvents, complex agents, and
high-pressure soil wash (Chu and Chan 2003; Conte et al. 2005). Use of this
technology is normally ex situ, but there are also some cases of in situ appli-
cation, depending principally on soil characteristics.
When determining in situ or ex situ remediation possibilities, organic
matter content and soil permeability play a key role. Indeed, for soil with
organic matter content <5%, in situ remediation technologies are applica-
ble; for organic matter >5%, ex situ technologies should be applied. In cases
of intermediate organic matter values, the pollutants adsorption onto soil
 Remediation of Contaminated Sites 13

will be the limiting mechanism determining whether an in situ or ex situ

technology should be applied (Okx and Stein 2000).
Moreover, a permeability factor (K Kf) > 0.2 m/d allows in situ remediation,
and ex situ technologies are most suitable for Kf < 0.2 m/d. An ex situ bio-
logical treatment is also feasible, but it will depend particularly on the soil
properties and the needed remediation time. In terms of time duration, ex
situ remediation processes last around 1 yr, whereas the remediation time
ranges normally between 1 and 5 yr years for in situ technologies (Bezama
et al. 2004).

B. Groundwater
Since the late 1970s, the on-site “pump and treat” process has been used
for groundwater remediation. Groundwater is pumped, treated normally in
afifilter/adsorbent system, and returned to the groundwater layer (Voudrias
2001). Sand filters,
fi granular activated carbon adsorbents, biofi filters, and
advanced oxidation processes have commonly been used as treatment
systems. The main disadvantages of this technology are related to its high
operational costs and the use of water as the carrier fluid. Still, the tendency
is toward increased use of in situ groundwater remediation.
In situ bioremediation has potential to remove contaminants present in
the groundwater plume, but it is limited by low ambient temperatures from
the aquifers. Mineralization processes from the contaminants could be seri-
ously affected by this factor, and biological on site treatment of contami-
nated groundwater could even require heating (Valo et al. 1990). There has
been very little experience with in situ biological treatment of contaminated
groundwater, as ex situ systems are more controllable, predictable, and less
complex than in situ treatments (Hamby 1996).
Activated carbon, minerals, waste-derived adsorbents, and zero-valent
metals are the adsorbent materials most used, with an interesting applica-
tion as a reactive wall for contaminant compound adsorption. Activated
carbon is mainly used in drinking and wastewater treatment facilities as
well as in groundwater remediation, and it can adsorb a wide range of
organic and inorganic pollutants present in water systems (USEPA 1998).
Its amphoteric character governs adsorption onto activated carbon. In fact,
in response to pH changes, the carbon surface develops coexisting electric
charges of opposite sign, whose prevalence depends on the solution’s chem-
istry. Therefore, attractive or repulsive electrostatic interactions between
the adsorbate and the adsorbent must be taken into consideration (Radovic
et al. 2000).
Activated carbon can adsorb heavy metals such as Cr(III), Cr(VI), Mo,
Co, Ni, Cu, Zn, Cd, Hg, Pb, U, Au, and As as well as phosphates (Radovic
et al. 2000), phenols (Streat et al. 1995), substituted phenols and benzenes
(Mollah and Robinson 1996; Aksu and Yener 2001), dyes (Walker and
Weatherley 1999), natural organic matter (NOM) (Radovic et al. 2000),
14 A. Bezama et al.

and other organics such as trihalomethanes, amines, chlorinated organic

compounds, alcohols, carboxylic and fatty acids, and amino acids (Radovic
et al. 2000). Because of its wide range of adsorption spectra, activated
carbon is considered a non selective adsorption material, although the high
cost of this adsorption material [between 2.5 and 9.00 (US)/kg] has driven
the search for other cheaper and selectively adsorbing materials. Research
and development on the use of natural soils as abundant, cheap, and selec-
tively natural adsorbents has been mainly focused on special soils. Such
soils have been recently investigated for their pollutant adsorption and
remediation capacity. As they are normally present abundantly in the envi-
ronment, they could consequently present interesting cost advantages. His-
torically, the soil’s porosity and pore-size distribution were of interest,
principally for their effects on water retention and flflow, gas advection and
diffusion, and nutrient transport. With increased concern about environ-
mental pollution, soil pore structure is now recognized as a critical factor
in pollutant sorption. Sorption has an underlying infl fluence on pollutant
transport, chemistry, and biological activity (Navia 2004).
Soil is composed of individual minerals and organic matter grains that
are cemented together to form particles. In turn, these particles may
agglomerate to form higher-ordered structures (Yaron et al. 1996). In short,
the association of its mineral and organic parts controls a soil system’s
porosity where soil water has a strong effect. From the point of view of
potential interactions with various pollutants, the constituents of the soil’s
solid phase should be grouped according to their surface area. The fate of
pollutants is affected by all the components of the soil solid phase. Still, soil
constituents with low surface area could principally affect the transport of
the pollutants as solutes, as immiscible with water liquids, or as vapors. The
soil solid phase can also indirectly induce organic pollutant degradation in
the soil medium through its effects on the water/air ratio in the system and
consequently on the soil’s biological activity. The group of constituents with
high surface area controls pollutant transport, retention, and release on and
from the soil surface as well as their surface-induced chemical degradation
(Yaron et al. 1996).
Regarding waste-derived materials, extensive research regarding non-
conventional, low-cost adsorption materials has been undertaken mainly to
identify the possible reuse of some organic and inorganic wastes. Inorganic
waste materials act as pure adsorbents, whereas organic waste materials
could act as adsorbents as well as biosorbents because of the microbiologi-
cal flora present in them.
Fly ash from thermal power stations and dried activated sludge were
used to adsorb and biosorb 2- and 3-chlorophenol, respectively, with satis-
factory results (Aksu and Yener 2001). Furthermore, chlorophenols and
nitrophenols were found to adsorb strongly onto a cheap carbonaceous
material obtained from the waste slurry generated in fertilizer plants (Gupta
et al. 2000), while cheaper activated carbons based on coconut shell, wood,
 Remediation of Contaminated Sites 15

coal, straw, and tires were tested successfully for the removal of phenol and
p-chlorophenol from contaminated water (Streat et al. 1995).
Bagasse fly ash, a sugar industry waste, has been recently investigated
for the removal of some specific fi toxic and carcinogenic compounds such as
pesticides based on a chlorobiphenyl structure (Gupta and Ali 2001). This
same residue is also capable of removing Cd and Ni from wastewater
streams (Gupta et al. 2003).
Low-cost adsorbents obtained from organic residues have also been
developed to remove hexavalent chromium, which is often found in waste-
water discharges from electroplating, metal finishing, and chrome prepara-
tion, and is considered to be highly toxic with a potential carcinogenic
effect. At acidic pH, sawdust, sugar cane bagasse, sugar beet pulp, and corn
cob, which are naturally occurring cellulosic waste materials, are able to
adsorb Cr(VI) present in contaminated water (Sharma and Foster 1994).
Moreover, Cu and Cd ions have been found to adsorb successfully onto
bone char (Ko et al. 2000), whereas Al, Ca, Cd, Cu, Fe, Mg, Ni, Pb, and
Zn ions (especially Pb) were efficiently
fi removed from an acidic leachate
by cocoa shells and, to a lesser degree, by cedar bark (Meunier et al.
2002).
With respect to colored effl fluents, these wastewaters are not only aes-
thetically displeasing, but they also impede light penetration, thus upsetting
biological processes within a stream. Additionally, many dyes are toxic to
some organisms and may cause direct destruction of aquatic communities,
requiring some form of advanced treatment. Adsorption of acid and basic
dyes present in aqueous solutions onto low-cost adsorbents such as bagasse
pith, peat, corn cob, bean waste, and sugar-industry mud (Magdy and
Daifullah 1998), as well as onto low-cost sewage sludge-based activated
carbon (Rozada et al. 2003), have produced very successful results.
Zero-valent metals are one of the most promising adsorbent materials,
with an interesting application for pollutant adsorption in in situ ground-
water remediation, and are partially displacing on-site groundwater treat-
ment processes like “pump and treat.” In this process, the groundwater is
pumped, treated normally in a filter/adsorbent system, and returned to the
ground, although the tendency is toward increased use of in situ remedia-
tion (Zorzi and Hammer 1998). Fe0, Al0, Zn0, Ni0, Cu0, Pd0/C, and their
combinations have been successfully tested for organochloride removal
from groundwater. Moreover, it has been demonstrated that zero-valent
iron is capable of producing reductive dechlorination of a great spectra of
chlorinated organic compounds (Table 5).
Some unknown Fe0-treatable CHCs have recently been carefully studied.
Particularly, PCB reductive dechlorination may also occur in the presence
of zero-valent iron while extracting PCBs from soil and sediments with
subcritical water (Yak et al. 1999). From the chlorophenol group, the elec-
trochemical dechlorination of 4-chlorophenol to phenol was determined to
occur rapidly on palladized carbon cloth or palladized graphite electrodes
16 A. Bezama et al.

Table 5. Organochlorine compound treatability with Fe0 as reactive wall.

Treatable organochlorine compounds

Methanes Trichloromethane. tetrachloromethane
Ethanes 1,1-Dichloroethane; 1,1,2-trichloroethane;
1,1,1-trichloroethane; 1,1,1,2-tetrachloroethane;
1,1,2,2-tetrachloroethane; hexachloroethane
Ethenes Vinyl chloride; 1,1-dichloroethene; trans-1,
2-dichloroethene; cis-1,2-dichloroethene; trichloroethene;
tetrachloroethene
Propanes 1,2-Dichloropropane; 1,2,3-trichloropropane
Other Hexachlorobutadiene
Nontreatable organochlorine compounds
Methanes Chloromethane; dichloromethane
Ethanes Chloroethane; 1,2-dichloroethane
Organochlorine compounds with unknown treatability
Chlorobenzenes —
Chlorophenols —
Certain pesticides —
PCBs —

Source: Adapted from USEPA (1998); Gillham and O’Hannesin (1994).

(Cheng et al. 1997). At laboratory scale, complete dechlorination of penta-

chlorophenol was observed in a Pd/Mg reactive wall, while the dechlorina-
tion of less substituted chlorinated phenols by different metallic systems
was found to be more likely to be achieved (Morales et al. 2002).
Moreover, reductive dechlorination of lindane was achieved in aqueous
solution using a Zn-modifiedfi carbon cloth cathode (Kulikov et al. 1996).
Therefore, it is stated that in general zero-valent metal barriers could be
used for CHC dechlorination. The principal disadvantages of activated
carbon are its price and its consequent thermal regeneration, whereas Fe0
is an inexpensive material that should react and dechlorinate the CHCs
present in groundwater (Navia et al. 2002).

IV. Definitions
fi of an Integrated Approach
On May 19, 2005, Chile ratifified the Stockholm Convention to prevent the
damage caused by persistent organic pollutants (POPs) on the environ-
ment, including human health. With this ratifification, Chile agreed to apply
all the necessary measures to eliminate or restrict intentionally generated
POPs as well as to appropriately eliminate the existent unused POPs,
to minimize unintentional POP generation, and to establish a National
Stockholm Convention Implementation Plan. Within this framework, Chile
adopted as one of its priority lines the remediation of POP-contaminated
environmental liabilities.
 Remediation of Contaminated Sites 17

In Chile at present there is a generalized search for appropriate and

advanced soil and groundwater remediation technologies, where the inclu-
sion of regional conditions for this technology development is highly appre-
ciated (De Palma 2002; Bezama et al. 2007c). In site remediation, the ex
situ soil treatment plants are gaining importance in the European market
(Schmitz and Andel 1997) and also when dealing with chlorinated organics
(Deshpande et al. 1999; Khodadoust et al. 1999; Sheets and Bergquist 1999;
Chu and Kwan 2003; Haapea and Tuhkanen 2006). Although cost restraints
can be a factor, as mentioned earlier, in the case of CHC- and/or PCB-
contaminated soils and groundwater, a certain degree of accuracy in moni-
toring and controlling the handling and degradation of pollutants is required.
Therefore, in this work we propose an ex situ soil and groundwater reme-
diation scheme (Fig. 4) consisting of a soil washing process followed by a
further wastewater treatment with a filtration and an adsorption process.
In addition, characterization of the waste fractions is made to define fi the
possible fifinal treatment alternatives.

A. Soil Washing
First, contaminated soil should be sieved. The large size soil fraction, which
is normally less contaminated, should be immediately disposed. The con-
taminated soil fraction should be washed under optimal temperature, pH,
and tensoactive concentration. With these optimized values, the final pol-
lutant distribution in the different soil-size fractions should be determined.
It is expected that the pollutants should concentrate in the fine
fi soil fraction
<63 μm (Reinert et al. 1999), which will be characterized and treated further
in a windrow composting system. As guide values for the chlorinated organ-
ics concentration in cleaned soil, it is possible to use the Austrian Norm for
soil remediation S2088-2 (OeN 2000), where limit values and prevention
values are established (Table 6). For possible soil landfilling
fi after the clean-
ing process, the Austrian Landfi fill Policy (List 2001) should be applied, dif-
ferentiating between four landfillfi types, and each one has limit values for
pollutants in the soil (Table 7).

B. Wastewater Treatment
The wastewater stream from the soil washing process will be contaminated
with CHCs. This stream should be cleaned by adding fl flocculating com-
pounds through a press filter
fi and a further activated carbon adsorption
process. The filtration process will treat the suspended compounds, while
the activated carbon system should adsorb the dissolved pollutants. The
cleaned wastewater should be returned to the washing system as process
water, following an integrated process concept. The Austrian Water Law
will be used for fi
first-limit wastewater values. The emission limit values will
follow the indication for emission into open canalization (Table 8).
18 A. Bezama et al.

Fig. 4. Overview of the proposed integral concept for the remediation of organo-
chlorine compound-contaminated soils.
Table 6. Limit and prevention values for polyaromatic
hydrocarbons (PAH) and PCB presence in soil.

Limit value Prevention value

Pollutant (mg/kg dry soil) (mg/kg dry soil)

PAH 1 50
PCB 0.2 1

Source: OeN (2000).

 Remediation of Contaminated Sites 19

Table 7. Limit values for selected pollutants depending on landfill

fi type.

fill type (differentiated by the source of the wastes)a

Landfi

Excavated Demolition Residual Mass waste

soil (mg/kg waste (mg/kg waste (mg/kg (mg/kg dry
Pollutant dry mass) dry mass) dry mass) mass)

Total content
PAH 0.5 2 t.b.d. 100
Hydrocarbons 20 100 5,000 20,000
TOC (as C) 20,000 30,000 30,000 50,000
POX (as Cl) — — — 1,000
Elute
Hydrocarbons 5 50 100 —
TOC (as C) 200 500 500 t.b.d.
EOX (as Cl) 0.3 3 t.b.d. 30
Surfactants 1 5 20 t.b.d.
(as TBS)

t.b.d., to be determined if relevant; TOC, total organic carbon; POX, purgeable organic
halogens; EOX, extractable organic halogens.
a
Types of sanitary landfills
fi defi fined according to FEA (2001); Jacobsen and Kristoffersen
(2002).

Table 8. Wastewater limit values.

Organic pollutants Limit value (mg/L)

AOX (as Cl) 0.5

Nonvolatile lipophilic 100
compounds
Hydrocarbons 20
POX (as Cl) 0.1
Phenol index (as phenol) 10
BTX 0.1

AOX, adsorbable organic halogens; POX, purgeable

organic halogens; EOX, extractable organic halogens; BTX,
benzol, toluol, xylol.
Source: Rossmann (1993).

C. Characterization of Waste Streams

The different waste streams (soil fi
fine fraction, filtration steps, solid waste,
and spent activated carbon) will be tested for pollutant load level and bio-
degradability. The wastes with a pollutant load level below the Austrian
Landfifill Policy will be immediately landfi filled, and the biodegradable
20 A. Bezama et al.

Table 9. Limit values for pollutants in compost.

Pollutant Limit value (mg/kg dry mass)

PAH 2
PCB 0.2
Hydrocarbons 200

polluted fractions will be treated in a composting system. Possible reuse of

the cleaned soil fractions in the construction market as well as the reuse of
the stabilized material (from the windrow composting process) in recultiva-
tion purposes will be analyzed. For the nonbiodegradable fractions, a
thermal processing criterion will be developed, focused especially on use
at cement industry facilities.

D. Windrow Composting
Biological degradation of the organochlorine compounds should be per-
formed in a windrow composting system. The fractions to be handled are
the biodegradable, still-contaminated fine fraction from the soil washing
process, the spent activated carbon, and the waste fraction from the waste-
water treatment’s fi filtration process. To optimize the relationship between
the nutrients (i.e., carbon, phosphorus, and nitrogen) in the biological
process, structural material and appropriate substrates (i.e., amendments)
should be added. As a final product, a stabilized material is obtained that
should at least comply with the limit values presented in Table 8 for its
further landfilling.
fi To search for additional uses, a complete characteriza-
tion of the stabilized material shall be determined. For land applicability
uses, the compost limit values are regulated by the Austrian Compost
Policy as summarized in Table 9 (BLFUW 2001).

V. Discussion
Contaminated sites are becoming more important throughout the world, as
their containment and/or remediation can be coupled with a reasonable
further redevelopment of these sites, which is foreseen as one of the most
important future environmental tasks. In the future, research on contami-
nated site remediation must focus on the application of combinations of
already existing technologies in such a way as to appropriately degrade the
most abundant pollutants in an economically and environmentally sound
way. Particularly, organochlorine compounds represent an important frac-
tion of the pollutants found at the identified
fi contaminated sites in Europe,
especially considering those originating from industrial and agricultural
activities, being present in approximately 75% of the identified
fi potentially
contaminated sites. Therefore, the sites contaminated with such pollutants
 Remediation of Contaminated Sites 21

require innovative remediation solutions that make possible future reuse

of these derelict areas.
This work has focused on the three most representative groups of
organochlorine compounds, namely polychlorinated biphenyls (PCBs),
chlorinated hydrocarbons (CHCs), and chlorophenols (CPs). A complete
analysis has been established for the determination of an appropriate tech-
nology that could handle these compounds in an environmentally and tech-
nically sound manner but that could also be suitable for developing countries
and especially for Chile. Such a concept has been defi fined as an integral
approach to treat and manage the contaminated material that comprises
soil excavation and subsequent mechanical sorting of the material. Two
material streams can be defi fined from this process, where each one will
undergo a different treatment according to their pollution loads. The
defifined treatment options comprise mechanical sorting and soil condition-
ing and a soil washing process coupled with either biological or thermal
remediation, depending on the nature of the pollutants present in each
project. Final treatment options comprise the disposal of the material in a
sanitary/security landfi
fill but also consider the recycling of the material in
applications such as filling materials or for the recultivation of degraded
areas, an aspect that can be considered only on a case-by-case basis.
In the case of developing countries such as Chile, such technological
approaches could be easily implemented because they would not result in
technological adversities or barriers in the already existing market. The
technical concepts analyzed and presented in this review can be considered
as suitable for application regardless of the country’s economic situation
and thus provide an interesting option when looking for sound environment
recovery alternatives.

Summary
Chlorinated hydrocarbons represent an important aspect of the contami-
nated sites identifi
fied worldwide, especially when considering those origi-
nated from industrial or agricultural activities, which represent approximately
75% of the potentially contaminated sites identifi fied in developed European
countries.
Chlorinated hydrocarbons, such as trichloroethylene, perchloroethylene,
dichloromethane, 1,1,1-trichloroethane, or chloroform, are used extensively
in many industries (e.g., in metal degreasing, dry cleaning, paint industries,
organic synthesis, etc.), and their common use can make them into a sig-
nificant
fi pollution source in the atmosphere, soils, and aquifers. Another
important group of organochlorine compounds are polychlorinated
biphenyls (PCBs). Among the characteristics of the PCBs, one of the most
important is a wide variability in their physicochemical properties (vapor
pressure, water solubility, and partition coefficients)
fi because of their
great number of congeners (varying from very slight compounds, such as
22 A. Bezama et al.

monochlorobiphenyl, to the not very volatile compound decachlorobiphe-

nyl), which determines their behavior and mobility in different environ-
mental compartments.
Remediation can be defined
fi as the improvement of a contaminated site
to prevent, minimize, or mitigate damage to human health and/or the envi-
ronment. Remediation involves the development and application of a
planned approach that reduces the availability of contaminants to receptors
of concern. The selection of an appropriate technology is therefore a crucial
activity in a remedial project, where technical, economic, social, legal, and
environmental aspects must be considered and evaluated. Conventional
groundwater treatment methods (e.g., based on sorption onto granular
activated carbon) are in many cases not very effective, in particular for
some low sorption potential compounds. Permeable reactive barriers con-
structed of elemental iron have emerged as an effective passive remediation
method for groundwater contaminated with a range of contaminants,
mainly chlorinated hydrocarbons. Furthermore, soil washing is a state-of-
the-art technique that transfers the contaminants from the soil to a liquid
phase by desorption and solubilization. In these applications, extracting
solutions (typically water with or without additives to improve contaminant
removal) are infiltrated
fi into the soil, and the loaded aquifer is then either
treated in situ or recovered and treated on site. Nowadays there is a gen-
eralized search for appropriate and advanced soil and groundwater reme-
diation technologies whereby the inclusion of the regional conditions to this
technology development is highly appreciated.
The present work aimed to fi first present the technical aspects that need
to be considered for an appropriate selection of remedial technologies for
site remediation when dealing with organohalogenated compounds as the
main pollutants. From these aspects, an integral site remediation strategy
was defined,
fi based on the European experience but easily transferable to
the reality of developing countries in South America, specifi fically consider-
ing Chile. This strategy consists in the remediation of CHC-contaminated
soil through a soil washing process followed by a further wastewater treat-
ment consisting of a filtration and an adsorption process. In addition, a
characterization of the waste fractions will be made to defi fine the possible
treatment alternatives.

Acknowledgments
This work was partially supported by FONDECYT N° 1050647 (R. Barra),
and by The Research Directorate of the University of Concepción.

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Manuscript received March 22; accepted April 4, 2007.

Rev Environ Contam Toxicol 193:31–52 © Springer 2008

Chemistry and Fate of Triazolopyrimidine

Sulfonamide Herbicides

Thomas W. Jabusch and Ronald S. Tjeerdema

Contents
I. Introduction .......................................................................................................... 31
II. Chemistry .............................................................................................................. 31
A. Physicochemical Properties .......................................................................... 31
B. Synthesis .......................................................................................................... 36
C. Mode of Toxic Action ................................................................................... 36
III. Methods of Analysis ............................................................................................ 38
IV. Environmental Fate and Occurrence................................................................ 39
V. Photodegradation................................................................................................. 40
VI. Metabolism in Microorganisms, Plants, and Animals .................................... 43
Summary ............................................................................................................... 48
References ............................................................................................................ 48

I. Introduction
The triazolopyrimidine sulfonamide (TSA) herbicides were registered in
the United States in 1993. Their mode of action is by inhibition of aceto-
lactate synthase (ALS), an enzyme common to plants and microorganisms
but not found in animals. ALS inhibitors include other herbicide families
such as the sulfonylureas, imidazolinones, and pyrimidinyl thiobenzoates.
In the 1970s, sulfonylureas were the first
fi ALS inhibitors to be introduced
to the market. Their discovery was greeted as a great achievement because
of their ability to suppress weeds at extremely low application rates com-
pared to previously used herbicides and with low toxicity risk to humans
and wildlife. By 1991, the market value of ALS inhibitors had reached $1.3
billion (U.S.). However, some of the problems associated with their use
include the induction of resistance in weed species to both ALS-inhibiting
and alternative herbicides. ALS inhibitors have also been found to damage

Communicated by Ronald S. Tjeerdema.

Thomas W. Jabusch
San Francisco Estuary Institute, 7770 Pardee Lane, Oakland, CA 94621, U.S.A.
Ronald S. Tjeerdema
Department of Environmental Toxicology, College of Agricultural and Environmental
Sciences, University of California, One Shields Avenue, Davis, CA 95616-8588, U.S.A.

31
32 T.W. Jabusch and R.S. Tjeerdema

nontarget plants at residual levels that are below the detection limits of
standard analytical methods (Saari et al. 1994; Whitcomb 1999).
TSA herbicides were first developed in the late 1980s by Dow Agro
Sciences and are now widely applied for postemergence control of broad-
leaf weeds in food and forage crops (Table 1), commonly in rotation and/or
mixtures with different mode of action herbicides. Flumetsulam was the
first to be introduced to the U.S. market in 1993 to control weeds in corn
fi
and soybean fields.
fi It was followed by cloransulam-methyl and diclosulam,
which were developed for use with soybeans and peanuts for broadleaf
weed control. Although neither metosulam nor fl florasulam are registered
in the United States, they have been introduced in other countries for weed
control with maize (Kleschick and Gerwick 1989; Ware and Whitacre 2004;
PAN 2006). Penoxsulam, the newest addition, was registered in the U.S. in
2004 for the control of annual grasses, sedges, and broadleaf weeds in rice
culture (USEPA 2004a). At this writing, the new analogue pyroxsulam is
pending U.S. registration for control of annual grasses and broadleaf weeds
with winter wheat (USEPA 2006).

II. Chemistry
A. Physicochemical Properties
Triazolopyrimidine sulfonamides are among the newer structural types of
herbicides that have made important impacts upon the industry over the
past 10–15 yr; they represent an increasingly science-based approach to pest
control. They are selective ALS inhibitors, generally exhibit a broad activ-
ity spectrum with good crop selectivity and typically high herbicidal activity
at low application rates of 10–50 g active ingredient (a.i.)/ha (Kleschick
et al. 1992; Baskaran et al. 1996), and include seven compounds: flumet- fl
sulam, metosulam, cloransulam-methyl, diclosulam, florasulam,fl penox-
sulam, and pyroxsulam (Fig. 1). Their physicochemical properties are
summarized in Table 1.
Triazolopyrimidine sulfonamides exhibit acidic dissociation constants
(pKKas); thus, their water solubilities are highly pH dependent (see Table 1).
For example, the water solubility of penoxsulam ranges from 0.006 g/L at
pH 5 to 1.46 g/L at pH 9 (Roberts et al. 2003). Generally, these compounds
tend to be highly water soluble at basic and neutral pHs, and their solubility
decreases at lower pHs. Octanol–water partitioning and soil sorption coef-
ficients, as well as hydrolytic rates, for these compounds are also pH depen-
fi
dent. Log octanol–water partition coeffi ficients (log Kow) are low and indicate
that these agents do not signifi ficantly partition into lipids or other organic
solvent phases. The reported soil sorption coefficientsfi (Kds) for flumet-
fl
sulam (5–182) and penoxsulam (0.2–5.1) indicate moderate to high mobility
in soils (Kleschick et al. 1992; Jabusch and Tjeerdema 2005). Chemical
degradation by hydrolysis is also pH dependent; the hydrolytic half-life (t1/2)
Table 1. Physical, chemical, and toxicological properties of the TSA herbicides.

Cloransulam-
Property Flumetsulam Metosulam Diclosculam Florasulam methyl Penoxsulam Pyroxsulam

Chemical N-(2,6-
N N-(2,6-Dichloro-3- N-(2,6-
N N N
N-(2,6- Methyl 3-chloro- 2-(2,2- N-(5,7-
N
name Diflfluorophenyl)- methylphenyl)- Dichlorophenyl)- Diflfluorophenyl)- 2-(5-ethoxy-7- Difl fluoroethoxy)- Dimethoxy-[1,2,4]
5-methyl-[1,2,4] 5,7-dimethoxy- 5-ethoxy-7- 8-fl
fluoro-5- fluoro-[1,2,4] N
N-(5,8-dimethoxy- triazolo[1,5-a]
triazolo[1,5-a] [1,2,4]triazolo fl
fluoro-[1,2,4] methoxy-[1,2,4] triazolo[1,5-c] [1,2,4]triazolo[1,5-c] pyrimidin-2-yl)-2-
pyrimidine-2- [1,5-a] triazolo [1,5-c] triazolo[1,5-c] pyrimidine-2- pyrimidin-2-yl)-6- methoxy-4-
sulfonamide pyrimidine- pyrimidine-2- pyrimidine-2- sulfonamido) (triflfluoromethyl) (trifluoromethyl)
fl
2-sulfonamide sulfonamide sulfonamide benzoate benzenesulfonamide pyridine-3-
sulfonamide
Trade Accent gold Strongarm® Firstrate®, Gf-881®, Granite®,
name(s) herbicide®, Frontrow®, Grasp®
Broadstrike®, Gangster®,
Frontrow®, Gauntlet®,
Hornet®, Naf- Riverdale
280, Naf-281, 565®
Python®,
Scorpion®
Application Corn, soybean, Cereals Soybean, peanut Cereals, pasture Soybean Rice Wheat, hay, straw
Chemistry and Fate of Herbicides

barley, wheat
Application 10–80 g a.i./ha 30–70 g a.i./ha Maximum global Canada 5 g a.i./ha 17.5–44 g a.i./ha 40 g a.i./ha
rate use rate: 52 g Europe 7.5 g a.i./ha
a.i./ha
CAS 98967-40-9 139528-85-1 145701-21-9 145701-23-1 147150-25-4 219714-96-2 422556-08-9
number
Chemical C12H9F2N5O2S C14H13Cl2N5O4S C13H10Cl2FN5O3S C12H8F3N5O3S C15H13ClFN5O5S C16H14F5N5O5S C14H13F3N6O5S
formula
Molecular 325.3 418.3 406.1 359.3 429.8 483.4 434.4
weight
33
 34

Table 1. (cont.)

Cloransulam-
Property Flumetsulam Metosulam Diclosculam Florasulam methyl Penoxsulam Pyroxsulam

Vapor 4 × 10−9 Pa @ NA 7 × 10−10 Pa @ 25°C NA 4 × 10−14 Pa @ 9.5 × 10−14 Pa @ 25°C NA

pressure 25°C 25°C
pK
Ka 4.6 NA 4.1 NA 4.8 NA NA
Water 0.049 g/L at pH 2.5 NA 0.117 g/L at pH 5 NA 0.003 g/L at pH 5 0.006 g/L at pH 5 NA
solubility 0.124 g/L at pH 7 0.184 g/L at pH 7 0.41 g/L at pH 7
4.29 g/L at pH 9 1.46 g/L at pH 9
Log Kow 0.21 NA 1.42 at pH 5 NA 1.12 at pH 5 NA NA
−0.047 at pH 7 −0.365 at pH 7
−0.448 at pH 9 −1.24 at pH 9
Soil sorption 5–182 NA NA NA NA 0.14–5.05 NA
coefficient
fi
Kd
Soil t1/2 (d) 30–246 22 48 0.7–85 9–263 5.2–12.8 NA
Aqueous NA NA 101 NA 0.015 2.9–26.5 NA
photolysis
T.W. Jabusch and R.S. Tjeerdema

t1/2 (d)
Hydrolysis NA NA 1007 at pH 5 NA >365 at pH 5 NA NA
t1/2 (d) 241 at pH 7 118–231 at pH 7
1.9 at pH 9 3 at pH 9

Sources: Kleschick et al. 1992; Wolt et al. 1992; Frear et al. 1993; Baskaran et al. 1996; USEPA 1997; Parnell and Hall 1998; Jackson et al. 2000; Krieger et al. 2000a; Baron 2001; van
Weesenbeeck et al. 2001; Zabik et al. 2001; Rouchaud et al. 2002b; Roberts et al. 2003; USEPA 2004b; Borges et al. 2005b; Jabusch and Tjeerdema 2005; Cambridge Soft Corporation
2006; Jabusch and Tjeerdema 2006a; PAN 2006; Wood 2006.
 Chemistry and Fate of Herbicides 35

O O
N
N O
A. S NH
F B. N N
N S NH Cl
N O N
O N O
F
Cl

F
O Cl
C. D. N O
O
N N N S NH F
S NH Cl N N N
O
F N O F
O

O
F N O O
S NH O N N N
N N N NH O
E. F. N F
O S F
Cl O
O O F
O

F
F

F
H OF F
O N N
G. S
N
N O
N O N
O

Fig. 1. Chemical structures of the triazolopyrimidine sulfonamides. A. Flumetsulam

(N-(2,6-difluorophenyl)-5-methyl-[1,2,4]triazolo[1,5-
fl α]pyrimidine-2-sulfonamide.
B. Metosulam (N-(2,6-dichloro-3-methylphenyl)-5,7-dimethoxy-[1,2,4]triazolo[1,5-
N
α]pyrimidine-2-sulfonamide). C. Diclosulam (N-(2,6-dichlorophenyl)-5-ethoxy-7-
N
fl
fluoro-[1,2,4]triazolo[1,5-c]pyrimidine-2-sulfonamide). D. Florasulam (N-(2,6-
N
difl
fluorophenyl)-8-fl fluoro-5-methoxy-[1,2,4]triazolo[1,5-c]pyrimidine-2-sulfonamide).
E. Cloransulam-methyl (methyl 3-chloro-2-(5-ethoxy-7-fluoro-[1,2,4]triazolo[1,5-
fl c]
pyrimidine-2-sulfonamido)benzoate). F. Penoxsulam (2-(2,2-difluoroethoxy)-
fl
N-(5,8-dimethoxy-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)-6-(trifluoromethyl)
benzenesulfonamide). G. Pyroxsulam (N-(5,7-dimethoxy-[1,2,4]triazolo[1,5-
N a]
pyrimidin-2-yl)-2-methoxy-4-(triflfluoromethyl)pyridine-3-sulfonamide).
36 T.W. Jabusch and R.S. Tjeerdema

for diclosulam decreases from 1,007 d at pH 5 to 1.9 d at pH 9 (van

Weesenbeeck et al. 2001). Available vapor pressure data range from
4 × 10−9 Pa to 4 × 10−14 Pa, suggesting that the triazolopyrimidine sulfon-
amides are in general extremely nonvolatile (Kleschick et al. 1992; Wolt et
al. 1992; van Weesenbeeck et al. 2001; Roberts et al. 2003). Soil degradation
half-lives range from 0.7 to 246 d and photodegradation half-lives from
0.015 to 101 d (Kleschick et al. 1992; Wolt et al. 1992; USEPA 1997; Jackson
et al. 2000; Krieger et al. 2000a; van Weesenbeeck et al. 2001; Zabik et al.
2001; Roberts et al. 2003; Jabusch and Tjeerdema 2005, 2006a).

B. Synthesis
Flumetsulam, the first marketed TSA herbicide, is derived from the
reaction of 5-methyl-[1,2,4]triazolopyrimidine-2-sulfonyl chloride with
2,6-difl
fluoroaniline in pyrimidine. The sulfonyl chloride is prepared from
2-(benzothio)-5-methyl-[1,2,4]triazolo[1,5-α]pyrimidine by reaction with
chlorine in aqueous acid (Fig. 2; Kleschick et al. 1992). At near-neutral pHs,
the sulfonamide bridge of triazolopyrimidine sulfonamide molecules is
thought to be deprotonated, rendering the compound highly water soluble
(Renew and Huang 2004). At lower pHs, the ratio of the anionic to the
neutral species shifts toward the latter, and solubility as well as soil mobility
slightly decrease (Wauchope et al. 2002).

C. Mode of Toxic Action

The general mode of action of the TSA herbicides is the inhibition of ALS,
which catalyzes the first
fi common step in the biosynthesis of the branched-
chain amino acids (valine, leucine, and isoleucine) in plants and micro-

F
N O N O
N N
S Cl + H2N S NH F
N N N
N O N O
5-Methyl[1,2,4]triazolo F Pyridine F
[1,5-a]pyrimidine- 2,6-Difluoroaniline
Flumetsulam
2-sulfonyl chloride
N-(2,6-Difluorophenyl)-5-methyl-
[1,2,4]triazolo-
Cl2 AcOH-H2O [1,5-a]pyrimidine-2-sulfonamide
-5°C to 3°C

N N
S
N N

2-(Benzylthio)-5-methyl-
[1,2,4]triazolo[1,5-a]pyrimidine

Fig. 2. Reaction scheme for the synthesis of fl

flumetsulam.
 Chemistry and Fate of Herbicides 37

organisms (Subramanian et al. 1989). There are to date four classes of

ALS–inhibiting herbicides: the triazolopyrimidines, sulfonylureas, imidazo-
linones, and pyrimidyl-oxy-benzoates (Mourad and King 1992). They all
bind to closely overlapping sites on the ALS molecule that are presumably
separate from binding sites for substrates or feedback inhibitors such as
valine (Landstein et al. 1993).
The herbicidal activities of both flumetsulam
fl and metosulam were found
to be temperature mediated. Elevating temperature after spraying from 1°
to 20°C increased the activities of both fl flumetsulam and metosulam in the
broad-leaved weed Raphanus raphanistrum by factors of 97 and 7, respec-
tively, with 50% reduction rates (ED50) for flumetsulam
fl ranging between
19.5 (1°C) and 0.2 g a.i. ha−1 (20°C) and for metosulam between 0.55 (1°C)
and 0.08 g a.i. ha−1 (Madafiglio
fi et al. 2000).
ALS herbicide resistance has been observed worldwide, and the devel-
opment of target site resistance resulting from repeated applications is
probably the most signifi ficant barrier to the long-term successful use of TSA
herbicides (Gerwick and Kleschick 1991; Rubin 1996; Whitcomb 1999).
Almost all cases of resistance to TSA herbicides or other ALS inhibitors
are the result of an altered ALS enzyme that is less sensitive to inhibitors
(Schmitzer et al. 1993; Bernasconi et al. 1995). Resistance is commonly the
result of a point mutation within discrete conserved domains of the als gene,
resulting in an altered target site and, thus, target site resistance. There are
different possible mutations within the ALS system, conferring a broad
spectrum of resistance to ALS inhibitors (Shaner and Singh 1997; Sprague
et al. 1997). In addition, varying degrees of cross-resistance between triazo-
lopyrimidines, sulfonylureas, and pyrimidinyl-oxy-benzoates have been
detected in different plants (Landstein et al. 1993; Foes et al. 1998;
Ferguson et al. 2001; Hashem et al. 2001; Patzoldt et al. 2001). Patterns of
resistance and cross-resistance appear to be specifi fic to the particular point
mutation and its position on the ALS enzyme (Whaley et al. 2006). In most
instances, resistance has been inherited as a semidominant trait (Saari et
al. 1994; Boutsalis and Powles 1995; Devine and Eberlein 1997; Patzoldt
and Tranel 2002; Sibony and Rubin 2003; Zheng et al. 2005).
A different resistance mechanism, observed in Alopecurus myosuroides,
is the rapid detoxification
fi of the ALS-inhibiting herbicide (Kemp et al.
1990). Selectivity of flflorasulam in wheat has been determined to be primar-
ily related to a differential rate of metabolism between wheat (t1/2 of 2.4 hr)
and broadleaf weeds (t1/2 ranging from 19 to >48 hr) and, to a lesser extent,
to uptake differences (deBoer et al. 2006). Differences in cloransulam
absorption and translocation partially explained differences in susceptibil-
ity among some weed species but not others (Barnes and Oliver 2004).
Cloransulam and other ALS inhibitors have also been found to anta-
gonize graminicides. This antagonism has not been fully explained. Pro-
posed hypotheses suggest either a decline in graminicide absorption or
prevention of the inhibiting action of the graminicide on meristematic
growth (Barnwell and Cobb 1994).
38 T.W. Jabusch and R.S. Tjeerdema

III. Methods of Analysis

Because of the low use rates of TSA herbicides, ultratrace residue meth-
odologies are required for their detection in environmental samples or to
enforce tolerance levels in food crops (Maycock et al. 1995; Shackelford et
al. 1996; Whitcomb 1999). Analytical methods with high sensitivity levels
for the detection of TSA herbicides in environmental matrices include
liquid chromatography with ultraviolet detection (LC-UV), thermospray
liquid chromatography with mass spectrometry (TSP-LC-MS), capillary gas
chromatography with mass spectrometry (GC-MS), LC tandem MS (LC-
MS/MS), capillary electrophoresis with UV detection (CE-UV), coelectro-
osmotic CE-UV, and direct enzyme-linked immunosorbent assay (ELISA).
All these methods are to varying degrees associated with some inherent
difficulties
fi but are able to detect one or several TSA herbicides in the ppb
range after varying extraction, cleanup, and preconcentration procedures
(Maycock et al. 1995; Baskaran et al. 1996; Shackelford et al. 1996; Krynitsky
1997; Parnell and Hall 1998; Krieger et al. 2000b; Laganà et al. 2000; Borges
et al. 2005a–c).
LC-MS and CE were successfully employed for ultratrace analysis of
TSA herbicides in environmental matrices in multiresidue determinations
at the ppb range. CE is a high-efficiency
fi separation method for pesticide
analysis in complex environmental matrices, but its low detection sensitiv-
ity, which is normally in the ppm range, needs to be overcome; this can be
achieved by sample preconcentration with a method called sample stacking.
Krynitsky (1997) analyzed fl flumetsulam along with 12 sulfonylurea herbi-
cides in runoff water by CE and LC/MS and achieved a limit of quantitation
(LOQ) of 0.02 ppb with a sample preparation method that included acidi-
fication, extraction by reversed-phase SPE, and extract cleanup with a
fi
tandem sample stacking system consisting of a strong anion-exchange SPE
cartridge stacked on an alumina cartridge. For the determination of fi five
TSA herbicides (diclosulam, cloransulam-methyl, fl flumetsulam, metosulam,
and flflorasulam) in soy milk, Borges et al. (2005c) developed a methodology
combining SPE and CE-MS with sample stacking in normal stacking mode
(NSM) and achieved limits of detection (LODs) down to 74 ppb. The same
authors achieved LODs between 0.1 and 0.24 ppb for analysis of the
same suite of TSA herbicides (diclosulam, cloransulam-methyl, flumet- fl
sulam, metosulam, and florasulam) in water samples with a methodology
that employed CE-UV, SPE using C18 cartridges, and on-line preconcentra-
tion by sample stacking with matrix removal (SWMR; Borges et al. 2005b).
The separation electrolyte was 24 mM formic acid and 16 mM ammonium
carbonate at pH 6.4; recovery ranged between 55% and 110%. For the
detection of TSA herbicides in soils, the authors combined a methodology
using SPE (using C18 cartridges), on-line fi field-enhanced sample injection
(FESI), and coelectroosmotic CE-UV and achieved LODs between 18 and
34 ppb (Borges et al. 2005a).
 Chemistry and Fate of Herbicides 39

IV. Environmental Fate and Occurrence

The physicochemical properties and experimentally determined degrada-

tion rate constants of the TSA herbicides (see Table 1) indicate their mod-
erate to low persistence in the environment, which is confirmed
fi by available
field dissipation studies. For instance, penoxsulam applied to flooded and
fi
nonfl flooded rice plots in Arkansas, California, Italy, and Spain yielded first-
order t1/2s of 3–7 d (Roberts et al. 2003). Field dissipation of cloransulam-
methyl and diclosulam was best characterized by two-compartment
modeling, presumably resulting from multiple concurrent degradation
mechanisms (photolysis, hydrolysis, and aerobic soil metabolism; van
Weesenbeeck et al. 1997; Zabik et al. 2001). Field dissipation rates were
determined with initial t1/2s ranging from 2.5 to 4.8 d for cloransulam-methyl
(van Weesenbeeck et al. 1997) and from 13 to 43 d for diclosulam (Zabik
et al. 2001).
Dissipation of fl
flumetsulam in the 0–8 cm soil layer of cornfi field soils also
followed first-order kinetics with t1/2s of about 41 d for crops grown on
sandy-loam soil and 30 d for crops grown on loamy-sand soil (Rouchaud
et al. 2002a). Flumetsulam degradation t1/2s in five Mississippi soils of
varying texture (loam, silt loam, and clay loam), organic matter content
(1.2%–3.5%), and pH (6–7.6) were ranging between 20 and 46 d (Shaw
and Murphy 1997). In this study, fl flumetsulam persistence decreased with
increasing cumulative rainfall and decreasing organic matter content
(within a range of 1.2%–3.5%). Soil pH had no measurable effect (Shaw
and Murphy 1997). Lehmann et al. (1992) observed similar degradation
rates in flumetsulam but found a dependency on both sorption and pH in
21 U.S. soils. They described degradation rates with a first-order
fi model,
using organic carbon and pH to estimate sorption from Koc values for the
neutral and anionic forms of fl flumetsulam (Lehmann et al. 1992). At appli-
cations rates of 50–100 g a.i. ha−1 on corn, fl
flumetsulam residues in soils gave
injury syndromes to sensitive cabbage crops planted 1 yr later (O’Sullivan
et al. 1999). Also, there were no, or only very light, phytotoxicity symptoms
observed in sensitive crops (sugar beet, lettuce) sown 5 mon after applica-
tion of flumetsulam at a low application rate of 20 g a.i. ha−1. Flumetsulam
is used in the U.S. at the rate of 60–80 g a.i. ha−1 for the control of broad-
leaved weeds and grasses in corn and soybean culture (Rouchaud et al.
2002b).
Soil mobility varies between different TSA herbicides. Residues of
penoxsulam in soil were low and transient after application to flooded
fl and
nonfl flooded rice fields (Roberts et al. 2003). Cloransulam-methyl was
retained in the soil surface layer (0–15 cm) after application to soybean
crops despite strong leaching conditions at test sites (van Weesenbeeck et
al. 1997). Modeling results with the PRZM 2 (Pesticide Root Zone Model-
ing) model suggest that cloransulam-methyl is potentially mobile in finer- fi
texture soils with <1.5% organic matter. Flumetsulam concentrations in
40 T.W. Jabusch and R.S. Tjeerdema

three Mississippi soils of varying texture and organic content were primarily
limited to the upper 8 cm of soil 84 d after treatment (Murphy and Shaw
1997). During the cropping period, fl flumetsulam residues were detectable
but low (near the GC-MS method detection limit of 0.3 ppb) in the 8–15 cm
surface layer and were never detected in the 15–20 cm layer. The results
indicate that fl
flumetsulam strongly adsorbed to soil and that leaching is not
a signifificant dissipation route (Rouchaud et al. 2002a). The reported soil
t1/2 of fl
flumetsulam ranges from 30 to 60 d at pH 6–7 (2%–4% organic
carbon; Gerwick and Kleschick 1991).
A runoff study combined with PRZM modeling revealed that less than
6% of diclosulam applied to peanut fi fields would be lost from severe rain-
fall, assuming worst case management practices. Of the total diclosulam
runoff, 97% was transported off the field by water, with <3% associated
with the sediment (van Weesenbeeck et al. 2001). Soil aging effects,
resulting in increased adsorption, can contribute to the limited mobility
of diclosulam (Yoder et al. 2000; Zabik et al. 2001), cloransulam-methyl
(van Weesenbeeck et al. 1997), and florasulam in soils (Krieger et al.
2000a). The leaching potential of cloransulam-methyl to groundwater was
modeled using conservative inputs for soil, weather, and application rate.
The maximum predicted concentration in groundwater for the most vul-
nerable soil was 2.7 μg/L (ppb), which is more than two orders below
the U.S. EPA-defi fined health level of 1,000 ppb (USEPA 1997; van
Weesenbeeck and Havens 1999). The same study also suggests no signifi- fi
cant risk for nontarget terrestrial plants, based on comparisons of esti-
mated environmental concentration (EEC) and EC25 distributions obtained
from conservative Tier 2 phytotoxicity studies (van Weesenbeeck and
Havens 1999).
So far, few monitoring studies have attempted to determine ambient
TSA herbicide concentrations. In 1998, a monitoring study was conducted
to determine the occurrence of ALS inhibitors in rivers, reservoirs, and
groundwater in the midwestern U.S. Flumetsulam, the only TSA herbicide
measured, was detected in 83% of samples collected from 75 surface water
(63%) and 25 groundwater sites (12%), at a median concentration of
0.01 ppb and a maximum of 0.035 ppb. These concentrations are not likely
to be toxic to nontarget aquatic plants or of concern for human consump-
tion, but do add to the overall burden of pesticides carried by midwestern
rivers (Battaglin et al. 2000).

V. Photodegradation
Photolytic processes are considered important contributors to the degrada-
tion of TSA herbicides, both on soil and, particularly, in water (USEPA
1997; van Weesenbeeck et al. 1997; Krieger et al. 2000c; Zabik et al. 2001;
USEPA 2004b; Jabusch and Tjeerdema 2006b). Photodegradation studies
 Chemistry and Fate of Herbicides 41

with florasulam yielded a t1/2 of 3.3 d in a natural water system in summer

at 51.5°N latitude compared to an estimated t1/2 of 36 d based on photolysis
experiments in sterile buffered solution. The much faster rate in the natural
system suggests an important role for indirect photolytic processes in the
overall photodegradation of florasulam (Krieger et al. 2000c). In contrast,
degradation studies for penoxsulam in summer sunlight at 39°N yielded no
significant
fi differences between the rates determined in sterile buffered
solution and rice field water. Degradation t1/2s ranged from 3.3 to 3.7 d in
field waters of different origin compared to a t1/2 of 3.9 d determined in
fi
sterile buffered solution, indicating a minor role for indirect photolytic
processes in controlling the overall degradation process and rate
(Tjeerdema et al. 2005).
Proposed photodegradation pathways have been published for flora- fl
sulam (Krieger et al. 2000c; Fig. 3) and penoxsulam (Jabusch and
Tjeerdema 2006b; Fig. 4). In studies using radiolabeled florasulam
fl in
aqueous buffer, only one product (triazolopyrimidine sulfonic acid, TPSA;
Fig. 3) was found at greater than 10% of the applied radioactivity (it
attained 18% after 32 d; Krieger et al. 2000c). Analogous TPSA degradates
were also found as products of aqueous photolysis for diclosulam (Concha
et al. 1994) and penoxsulam (Fig. 4, pathway 2; Jabusch and Tjeerdema
2006b). However, the TPSA degradates were not detected in experiments
using natural water systems (Krieger et al. 2000c; Jabusch and Tjeerdema
2006b).
Photodegradation of florasulam in natural waters is proposed to proceed
by two main routes: (1) formation of 5-hydroxy fl florasulam by oxidation of
the methoxy group on the triazolopyrimidine ring to a hydroxyl group, or
(2) loss of the difluorophenyl
fl moiety by cleavage of the phenyl carbon–
nitrogen bond, followed by conversion of the methoxy group of the triazo-
lopyrimidine ring to a hydroxyl group (Krieger et al. 2000c). Subsequent
degradation of 5-hydroxy florasulam is by either (a) opening of the pyrimi-
dine ring and subsequent loss of the difluorophenyl
fl moiety, or (b) loss of
the difl fluorophenyl moiety first, followed by opening of the pyrimidine
ring.
Degradation studies with 14C-labeled penoxsulam in a “merry-go-round”
reactor suggest that aqueous photodegradation proceeds via three path-
ways: (1) cleavage of the sulfonamide bridge, (2) stepwise degradation of
the triazolopyrimidine system and its substituents, and (3) photooxidation
of the sulfonyl group. Seven major photoproducts were found (maximum
levels ≥5% during the course of the study) and six were identifi fied (see Fig.
4). Two of the identified
fi photodegradation products, 2-ATP and TPSA,
were present as 1.4% and 8.9%, respectively, of the initially applied radio-
activity. However, these compounds are assumed to be either rapidly bio-
degraded when formed or not formed in significant fi amounts in natural
waters (Krieger et al. 2000c; Jabusch and Tjeerdema 2006b).
42 T.W. Jabusch and R.S. Tjeerdema

F
N O
S NH F
N N N
O
O F

florasulam

F F F
N O N O O
N
S OH S NH2 S NH F
N N N N N N N N N
O O O
O O OH F

TPSA ASTP 5-OH-florasulam

F OH
O O N O
N
S NH2 S NH F
N N N N N
O O
H
F
OH

5-OH-ASTP DFP-ASTCA

OH

O N O
S NH2
N N
O
H

ASTCA
Fig. 3. Suggested photodegradation pathways for fl
florasulam in water (adapted
from Krieger et al. 2000c).
 Chemistry and Fate of Herbicides 43

O O O

N N 1 N
N
NH O N N N NH2
NH2 N N N
N S F F
O N
O F O OH
O

2-amino-TP 5-OH-2-amino-TP
F
F 2
penoxsulam

O
3
N N N
NH
O
N S
O
O OH
F F
F F TPSA
F F
O O
S NH S NH
O O N
N F O
F O N N
NH NH

F F
O HO
O O
BSTCA
BSTCA-methyl
F
F
F F
F O
F
O S NH
2
S NH ? O
O
O N
O N F
NH
F
F
F BST (sulfonamide)

Fig. 4. Suggested aqueous photodegradation pathways for penoxsulam: (1) cleav-

age of the sulfonamide bridge, (2) photooxidation of the sulfonyl group, and
(3) stepwise degradation of the triazolopyrimidine system and its substituents.
BSTCA, 3-(2-(2,2-difluoroethoxy)-6-(trifl
fl fluoromethyl)pheuylsulfonamido)-1H-1,2,4-
triazole-5-carboxylic acid (Adapted from Jabusch and Tjeerdema 2006b.)

VI. Metabolism in Microorganisms, Plants, and Animals

Aerobic soil metabolism of cloransulam-methyl, florasulam,
fl and diclosulam
is shown in Fig. 5. Degradation rates of the TSA herbicides and their deg-
radation products appear to be strongly influenced
fl by temperature, as well
as to some degree by soil moisture content (Kleschick et al. 1992; Wolt
et al. 1992, 1996; van Weesenbeeck et al. 1997; Cupples et al. 2000; Zabik
et al. 2001; Rouchaud et al. 2002b). For example, soil t1/2s of flumetsulam
44 T.W. Jabusch and R.S. Tjeerdema

A. F O
N O
F N O
S NH O
N N N S NH O
O N N N
Cl O O
O Cl HO
OH

cloransulam-methyl 5-OH-cloransulam-methyl

F N O O F O O
N
S NH O S NH OH
N N N N N N
O O
O Cl Cl
OH

cloransulam 5-OH-cloransulam

B. F
F
N O OH
N O
S NH F O
N N N S NH F O N
O N N N S NH F
F O N N
O F O
OH H
F
florasulam 5-OH-florasulam
(minor route)
OH
O
HN N N O
S NH F O
N S NH2
O N N
F O
H

C. ASTCA
O Cl (minor route)
O
N N N
S NH Cl
O
F N O HN N
S NH2
diclosulam N O
TSA
Cl
F O Cl
F N O N
S NH Cl F N O
S NH2
N N N N N N S NH Cl
O O N N N
OH Cl O
O
OH Cl
ASTP
5-OH-diclosulam 8-Cl-diclosulam

Fig. 5. Aerobic metabolism of triazolopyrimidine sulfoanimide (TSA) herbicides

in soil. A. Cloransulam-methyl (adapted from Wolt et al. 1996). B. Florasulam
(adapted from Jackson et al. 2000). C. Diclosulam (adapted from Yoder et al. 2000).
ASTP, aminosulfouyl triazdopyrimidine.
 Chemistry and Fate of Herbicides 45

in laboratory studies (both at 26°C) were found to range from 49 to 246 d,

whereas those of fl florasulam ranged from 1 to 8.5 d at 20°–25°C, and from
6.4 to 85 d at 5°C (Rouchaud et al. 2002b); the t1/2 of its major degradation
product, 5-hydroxy florasulam, ranged from 8 to 36 d at 20°–25°C and 43 to
78 d at 5°C (Krieger et al. 2000a). Dissipation t1/2s of diclosulam ranged from
16 to 54 d in U.S. and South American soils incubated in the dark at 20°C
and 45% moisture-holding capacity and at 20°C at 75% 0.3 bar moisture
(Yoder et al. 2000). Three soil metabolites reached levels of ≥10% of
applied compound (5-hydroxy diclosulam, aminosulfonyl triazolopyrimi-
dine (ASTP), and 8-chloro-5-hydroxy diclosulam). The terminal products
were CO2 and bound soil residues. The KD values of the metabolites were
slightly lower than that for diclosulam, and the sorptivity of both diclosulam
and its metabolites were found to increase over time (Yoder et al. 2000).
The degradation kinetics of cloransulam-methyl in tubes simulating
aerobic soil systems was observed to follow a biphasic pattern with a fast
initial step followed by a slower step. In this study, analyzed metabolites
had lower KDs and were less phytotoxic than the parent compound (Wolt
et al. 1996). However, in a different study the influence
fl of temperature on
the mineralization rate (conversion to CO2) was dependent upon the loca-
tion of the radiolabel within the cloransulam-methyl molecule, suggesting
that distinct groups of microorganisms degrade different parts of the mol-
ecule at different temperatures (Cupples et al. 2000). Also, mineralization
rates for cloransulam-methyl were found to be moisture dependent, with
direction of response (increase versus decrease) depending on soil type
(Cupples et al. 2000).
The t1/2 of penoxsulam in flasks simulating flooded rice field conditions
(anaerobic conditions but open to the atmosphere) ranged from 2 to 13 d;
faster degradation rates occurred in flasks with steeper redox gradients
(Jabusch and Tjeerdema 2006a). The suggested anaerobic pathway for
penoxsulam is shown in Fig. 6. In a strongly anaerobic test system (sulfate-
reducing or methanogenic) isolated from the atmosphere, flumetsulam
fl was
transformed abiotically to fl flumetsulam hydrate with a t1/2 of 186 d. The
transformation was reversed upon aeration, and the t1/2 of flumetsulam
hydrate was 2 d versus 40 d for fl flumetsulam (Wolt et al. 1992).
In plants, TSA herbicides are metabolized via para-hydroxylation of the
aniline ring, followed by conjugation with glucose (Fig. 7; Frear et al. 1993;
deBoer et al. 2006). Microsomal cytochrome P450-dependent monooxy-
genases were identifi fied as the enzymes responsible for flumetsulam hydrox-
ylation (Frear et al. 1993). Metabolism of florasulam
fl by broadleaf weeds
was so slow that metabolite characterization was not possible, but compari-
son of HPLC data suggests hydroxylation as the major pathway (deBoer
et al. 2006). There is very little information on the metabolism of TSA
herbicides in animals. Lactating goats, when given a daily oral dose of
radiolabeled cloransulam-methyl for 5 consecutive d, rapidly excreted most
of the compound within 23 hr of receiving the last dose, with 99.9% of the
46 T.W. Jabusch and R.S. Tjeerdema

O
O
N N N
NH O 2 NN N
NH2
N S F F N
O O
O F
O (Minor product)
2-amino-TP
F
F

Penoxsulam

1
O
N
NH O
HN N N F
S F
O
O F
O

F HN N
O
F
N NH
HO
5-OH-penoxsulam O S O
F
O F
F
F F

BSTCA
HN N
O
N NH
O
O S O
F
O F
F
F F

BSTCA-methyl HN N

N NH
F O S O
F
H2N
O O F
F O S F F
F F
O
F
F
BST

Sulfonamide
Fig. 6. Suggested anaerobic metabolism of penoxsulam in fl
flooded rice field soils.
(Adapted from Jabusch and Tjeerdema 2006a.)
 Chemistry and Fate of Herbicides 47

N O
N
S NH
F
N N O
F

flumetsulam

N O N O
N N
S NH F S NH F
N HO
N O N N O
F F

OH

OH

F
N N O
HO
S NH F
N N
O

N O
N HO OH N O
S NH F N
S NH F
N N O HO O
N N O
F O
HO OH F
HO
O OH
O
OH

F OH

N N HN H
HO OH
HO
SH F
N N
O O OH
O
OH

Fig. 7. Suggested metabolism of flumetsulam

fl in tolerant corn, wheat, and barley
seedlings by hydroxylation and glucose conjugation pathways (adapted from Frear
et al. 1993).

recovered radioactivity appearing in the urine and feces. Only one metabo-
lite, cloransulam, was identified,
fi amounting to 9.5% of total radioactive
residue in the liver (Lewer et al. 2000). Cloransulam is the corresponding
acid formed by ester hydrolysis of cloransulam-methyl (see Fig. 6A).
48 T.W. Jabusch and R.S. Tjeerdema

Summary
The TSA herbicides are generally applied to agricultural fi fields at low rates.
Possessing low mammalian toxicity, they pose little risk to either humans
or wildlife. Plants demonstrate a wide range in sensitivity to these com-
pounds. For example, the observed toxicity of fl flumetsulam to five different
aquatic plant species varied by more than 10,000 fold (Peterson et al. 1994),
beginning in the ppb range. Application of the TSA herbicides may pose a
potential risk to both aquatic and terrestrial plants at the manufacturer-
proposed rates, including the risk of injuring rotational crops by residues
of previous applications. There has been little ambient monitoring for the
TSA herbicides, but a study of selected low use rate herbicides in Midwest
U.S. waters found that observed fl flumetsulam concentrations were not
likely to be toxic to nontarget aquatic plants (Battaglin et al. 2000). Most
TSA herbicides possess low mobility but moderate persistence in soils, with
persistence increasing at lower temperatures. Environmental half-lives
range from less than a day to several months. Penoxsulam, which is desig-
nated for use with rice, is highly mobile but not very persistent in both water
and soils. Degradation of the TSA herbicides in the environment occurs by
a combination of processes (photodegradation, microbial degradation, and
hydrolysis). A major issue concerning use of the TSA herbicides is the rapid
development of herbicide resistance in target species. There are major data
gaps concerning the ambient fate and distribution of these compounds, and
it is undetermined whether, and if so, to what extent, TSA herbicides have
indirect ecological effects by contributing to the overall burden of xeno-
biotic substances in the environment.

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Manuscript received April 12; accepted April 18, 2007.

Rev Environ Contam Toxicol 193:53–210 © Springer 2008

Parameters for Carbamate Pesticide QSAR and

PBPK/PD Models for Human Risk Assessment

James B. Knaak, Curt C. Dary, Miles S. Okino, Fred W. Power,

Xiaofei Zhang, Carol B. Thompson, R. Tornero-Velez, and
Jerry N. Blancato

Contents
I. Introduction ..................................................................................................... 54
II. Nature of Carbamate Insecticides ................................................................ 56
A. Discovery ................................................................................................... 57
B. Carbamates and Their Toxicity .............................................................. 60
III. QSAR Models for Predicting Biological Parameters Used in
PBPK/PD Models ............................................................................................ 62
A. Human Oral Bioavailability .................................................................... 63
B. Toxicity Models ......................................................................................... 64
C. Liver Microsomal P450 and CYP Hydroxylation Models .................. 66
D. Tissue:Blood Partition Coefficients fi ........................................................ 66
IV. Experimentally Derived Biological Parameters Used in
PBPK/PD Models ............................................................................................ 68
A. Gastrointestinal Absorption .................................................................... 68
B. Skin Absorption ........................................................................................ 68
C. Tissue Partition Coeffi ficients/Distribution ............................................. 71
D. Glucuronidation and Transcellular Transport ...................................... 82
E. Metabolic Enzymes, P450s, and CaEs ................................................... 83
F. Models for In Vivo Metabolism of Carbamates ................................... 87
G. Response: In Vivo AntiChE Activity .................................................... 94
V. Target Enzymes, the ChEs and CaEs .......................................................... 98
A. Structure, Multiple Forms, and Distribution ........................................ 98
B. AChE, BChE, and CaE Substrate Selectivities ................................. 102

Communicated by James B. Knaak.

J.B. Knaak
Department of Pharmacology and Toxicology, School of Medicine and Biomedical Sciences,
SUNY at Buffalo, 3435 Main Street, Buffalo, NY 14214, USA.
C.C. Dary · M.S. Okino · F.W. Power
U.S. Environmental Protection Agency, Human Exposure and Atmospheric Sciences
Division, 944 E Harmon Ave, Las Vegas, NV 89193-3478, USA.
X. Zhang · C.B. Thompson
General Dynamics Information Technology, 181 N Arroyo Grande Blvd, Suite 105,
Henderson, NV 89074-1624, USA.
R. Tornero-Velez · J.N. Blancato
U.S. Environmental Protection Agency, National Exposure Research Laboratory, Mail Code:
D305-01 Research Triangle Park, NC 27711, USA.

53
54 J.B. Knaak et al.

VI. Acylation and Decarbamylation of Cholinesterases and CaEs .............. 109

A. Hydrolysis of Substrates ........................................................................ 109
B. Carbamylation (Inhibition) and Decarbamylation of ChEs
and CaEs .................................................................................................. 112
C. Bimolecular Rate Constants, ki, for the Ten Carbamates ................ 114
VII. Discussion ....................................................................................................... 118
VIII. Recommendations ......................................................................................... 122
Summary ......................................................................................................... 122
Acknowledgments ......................................................................................... 123
References ...................................................................................................... 123
Appendix A: Chemical Structures, Physical Parameters,
and Tissue Partition Coeffi ficients of Parent Carbamates
and Metabolites ............................................................................................. 142
Appendix B: Metabolic Pathways and Preliminary Metabolic
Rate Constants (Vmax, Km) for the Metabolism of Parent
Carbamates and Metabolites ....................................................................... 187
Appendix C: Physiological Model for Aldicarb Depicting Its
Metabolic Pathway ........................................................................................ 207
Appendix D: Nomenclature ........................................................................ 208
i. Acronyms and Abbreviations ............................................................ 208
ii. Chemical and Mathematical Expressions ......................................... 210

I. Introduction
Our interest in providing parameters for the development of quantitative
structure physiologically based pharmacokinetic/pharmacodynamic
(QSPBPK/PD) models for assessing health risks to carbamates (USEPA
2005) comes from earlier work with organophosphorus (OP) insecticides
(Knaak et al. 2004). Parameters specific fi to each carbamate are needed in
the construction of PBPK/PD models along with their metabolic pathways.
Parameters may be obtained by (1) development of QSAR models, (2)
collecting pharmacokinetic data, and (3) determining pharmacokinetic
parameters by fitting to experimental data. The biological parameters are
given in Table 1 (Blancato et al. 2000).
The quantitative structure-activity relationship (QSAR) model approach
requires (1) knowledge of the numerical values of the parameters of related
chemicals and (2) specification
fi of an equation for conducting the multiple
regression analysis (Fouchecourt et al. 2001). TOPKAT (Accelrys Software
Inc., San Diego, CA) is an excellent example of QSAR model development.
The program consists of several toxicity QSAR models; one provides infor-
mation on the acute oral toxicity of OP and carbamate insecticides. QSAR
models predicting Vmax and Km values for human liver microsomes (HLMs)
and CYP3A4 catalyzed hydroxylation reactions involving drug-based aro-
matic and alicyclic rings and aliphatic groups were recently developed by
Enslein et al. (2007). Liu et al. (2005) developed a QSAR model for pre-
dicting tissue/blood partition coeffi ficients, and Poulin and Theil (2002a)
 Parameters for Carbamate Models 55

Table 1. Biological Parameters Required for Carbamate Pesticide Physiologically

Based Pharmacokinetic/Pharmacodynamic (PBPK/PD) Models.a

Biological process Compartment Parameter Units

Absorption GI tract hr−1

Respiratory tract Blood : air PC Unitless
Skin Permeability cm hr−1
constant
Distribution Desired Blood : tissue PC Unitless
compartments
Metabolism, P450 Liver V bmax μmol hr−1 kg−1 bwc
cytochromes, etc. Kdm μmol kg−1
Excretion Kidney V bmax μmol hr−1 kg−1 bwc
Kmd μmol kg−1
Toxicity Blood, brain, liver B esterase M−1 min−1
inhibition hr−1
Recovery hr−1
Synthesis
a
Source: Blancato et al. (2000).
b
Maximum velocity.
c
bw, body weight.
d
Substrate concentration at half-maximum velocity.

used a mechanistic approach to predict tissue/blood partition coefficients.

fi
The Poulin and Theil (2002a) model was reexamined using log DpH 7.4 values
and was used to estimate partition coefficients
fi for the carbamates and their
metabolites.
A search of the literature for metabolism/pharmacokinetic data turned
up three carbamate reviews, written by Cool and Jankowski (1985), Kuhr
and Dorough (1976), and Roberts and Hutson (1999). The reviews consid-
ered chemical degradation, degradation in soils, and metabolism in plants
and animals, with metabolism in insects being covered by Kuhr and
Dorough (1976). Data in these reviews and others provided information
for metabolic fl flow charts or tables suitable for use in the development
of 10 preliminary carbamate PBPK/PD models using the Advanced Con-
tinuous Simulation Language (ACSL) (The AEgis Technologies Group,
Huntsville, AL).
A PBPK/PD model developed for isofenphos (CAS no. 25311-71-1)
(Knaak et al. 2002) was used as a template for the development of models
for aldicarb, carbaryl, carbofuran, formetanate, methiocarb, methomyl,
oxamyl, pirimicarb, propoxur, and thiodicarb. Bimolecular rate constants,
ki (pM−1 hr−1), were found describing the inhibition of the B-esterases [ace-
tylcholinesterase (AChE), butyrylcholinesterase (BChE), and carboxyles-
terase (CaE)] by the carbamates or their toxic metabolites. The relationship
between the inhibitory nature of the carbamates, structure of the target
56 J.B. Knaak et al.

enzymes (e.g., B-esterases), location of the B-esterases in tissues, and the

number of active sites/kg of tissue was reexamined using rate constants for
substrate hydrolysis (e.g., kcat). Metabolic rate constants, Vmax and Km, for
the cytochrome P450 (CYP450) hydroxylation of ring compounds, their
conjugation to glucuronic and sulfuric acid, and the hydrolysis of aryl and
oxime carbamates by hydrolases were obtained, if available, from the
literature.
No attempt was made in this review to obtain pharmacokinetic parame-
ters by fitting the output of preliminary ACSL-based PBPK models to
experimental data (e.g., absorption, distribution, metabolism, elimination,
and toxicity data). This approach was considered to be outside the scope
of the review. However, the metabolic pathways, molecular weights, tissue
partition coefficients,
fi bimolecular inhibition rates for AChE, ChE, CaE,
the number of active sites in tissues, metabolic Vmax, Km values, and other
parameters collected under approaches 1 and 2 were used by Zhang et al.
(2006, 2007), Okino et al. (2005a,b), and Power et al. (2005) with experi-
mental data and the Exposure Related Dose Estimating Model (ERDEM;
Blancato et al., 2006) to model carbofuran, carbaryl, and aldicarb.
Under the Food Quality Protection Act (FQPA) of 1996, the U.S. Envi-
ronmental Protection Agency (EPA) must consider available information
concerning the cumulative effects on human health resulting from exposure
to multiple chemicals that have a common mechanism of action, such as
the NN-methyl carbamates. ERDEM is capable of handling AMET (absorp-
tion, metabolism, elimination, and toxicity) of three or more carbamate
pesticides depending upon the total number of metabolites being modeled.
The upper limit of the current model (version 5.1) is 36 chemicals, which
includes parent pesticides and or metabolites (Miles Okino; U.S. EPA, Las
Vegas, NV).

II. Nature of Carbamate Insecticides

Carbamate insecticides are biologically active AChE inhibitors with the
general formula ROC(=O)NHCH3 for N-methylcarbamates and
ROC(=O)N(CH3)2 for dimethylcarbamates. The structure of the R group,
alkylphenyl, alkoxyphenyl, halogen phenyl, double ring and oxime, N,N- N
dimethyl, N-methyl,
N N-acyl, thiol (SC =O), thiono (OC =S), or dithio
N
(SC=S), and its effects on AChE activity and toxicity, are briefly
fl discussed
in the following section, “Discovery”. QSAR modeling played a very
important role in relating the structure of carbamates to their cholinester-
ase (ChE) inhibitory properties and insect toxicity under the World Health
Organization (WHO) Programme for the evaluation and testing of new
insecticides (Wright 1971). This program led to worldwide synthesis, testing
(adult mosquito, larval mosquito, and housefl fly), and the development of
structurally related carbamates to control vector-borne diseases with the
Departments of Entomology and Zoology, University of Illinois, Urbana-
Champaign, IL, playing a key role.
 Parameters for Carbamate Models 57

A. Discovery
Physostigmine
The fifirst known carbamate, physostigmine or eserine (CAS no. 57-47-6),
was discovered in extracts of the calabar bean. The alkaloid was found to
be miotic. Robertson (1863) suggested the drug for use in ophthalmology.
“Alkaloids are compounds of nitrogen, capable of uniting with acids, like
ammonia, and forming with them definitefi combinations, which are true
salts” (de Coninck 1886). The work of Stedman and Barger (1925) led to
the elucidation of the structure of physostigmine as the methylcarbamate
of a substituted indole derivative. Physostigmine appears to be the only
natural product within the N N-methylcarbamate group and is unstable in
water. The corresponding dimethylcarbamate (neostigmine, CAS no. 59-
99-4) was prepared by Aeschlimann and Reinert (1931) and was found to
be more stable and an effective miotic agent. Stemple and Aeschlimann
(1956) reviewed the properties of these compounds. Somani and Khalique
(1986) studied the distribution and pharmacokinetics of physostigmine in
rats after intramuscular administration. The time course of BChE activity
and plasma concentration showed that the maximum enzymatic inhibition
(47%) occurred about the same time (7 min) as the peak plasma concentra-
tion (583 ng mL−1 at 5 min) with enzymatic activity recovering to 81% at 2 hr
and 100% within 24 hr.

Dimethylcarbamates
Gysin (1954), at the Geigy Corporation, studied the insecticidal properties
of the dimethylcarbamates of 5,5-dimethyldihydroresorcinol (Dimetan,
CAS no. 122-15-6) and 1-iso-propyl-3-methyl pyrazole (Isolan, CAS no.
119-38-0). These two compounds, along with dimetilan (1-(dimethylcar-
bamoyl)-5-methyl-3-pyrazolyl dimethylcarbamate, CAS no. 644-64-4),
underwent considerable evaluation as commercial insecticides. All these
carbamates are potent anticholinesterase (antiChE) agents having I50 values
for AChE in fruit flies of 5–6 × 10−7 M for Dimetan to 7–10 × 10−8 M for
Isolan. The rate of hydrolysis of p-nitrophenyl methylcarbamate
(kb = 5.4 × 105 M−1 min−1) is about 107 times greater than that of the corre-
sponding dimethylcarbamate (kb = 3.4 × 10−2 M−1 min−1). The dimethylcar-
bamates are more stable than the methyl carbamates; however, as antiChEs
and insect toxicants, the methylcarbamates are far superior (Metcalf and
Fukuto 1962).

Phenylmethylcarbamates
Metcalf and March (1950) were among the first
fi to synthesize and study the
activity of simple phenyl methylcarbamates. Substitution in the ortho or
meta position with an alkyl group or halogen on the phenyl ring increased
insecticidal activity. Maximum activity was found with m-tert-butylphenyl
methylcarbamate. The mammalian/insect toxicity ratios ranged from >100
58 J.B. Knaak et al.

for butacarb (CAS no. 2655-19-8) to around 0.2 for aldicarb (CAS no. 116-
06-3). Increasing the chain length of N
N-alkyl groups exponentially increased
the recovery half-lives of inhibited insect AChE and decreased their inhibi-
tory potential when going from N N-methyl to N
N-ethyl groups (Yu et al.
1972a). The activity of alkoxyphenyl carbamates is governed by the same
set of rules that apply to the alkylphenyl carbamates. AntiChE and toxicity
for insects increase progressively with the size of the phenyl substituent,
with maximum activity at the isopropyl and sec-butyl groups. The ortho
positions are preferred over those of the meta positions. Goldblum et al.
(1981) developed a QSAR for a set of 269 phenyl N-methylcarbamates
N
inhibiting flfly head AChE (pI50 values). The data were taken from the
extensive studies of Metcalf and Fukuto (1962), Metcalf and Fukuto (1965a),
Metcalf and Fukuto (1965b), Metcalf and Fukuto (1967), and Metcalf et al.
(1965), which followed the earlier work of Metcalf and March (1950).

Halogen-substituted phenyl-, thio-, and nitrophenylcarbamates

The only halogen-substituted carbamate developed as a commercial insec-
ticide was o-chlorophenyl methylcarbamate, which was marketed in both
Japan and the United States (US). The electron-withdrawing properties of
these carbamates produce a substantial degree of hydrolytic instability.
According to Metcalf (1971), none of the thiocarbamates approach the
antiChE and insecticidal activity of the corresponding carbamate, indicating
that these compounds are very different from the phosphorothiolates [e.g.,
Gutoxon (CAS no. 961-22-8), Amiton (CAS no. 78-53-5), etc.]. Replacing
the carbamate linkage (OC (O) N) with thiol (SC=O), thiono (OC=S),
or dithio (SC=S) reduced the antiChE activity of 2-isopropylphenyl
N,N-dimethylcarbamate
N by 12 to 40 times (Petrovskii 1970).
Fifty-nine new N N-methyl and N,N-dimethylcarbamates,
N meta-
acylamidophenyl and meta-thioureidophenyl, were synthesized and tested
as inhibitors of fly and bovine ChEs (Sacher and Olin 1972). Structure–
activity correlation indicated that maximal antiChE activity was associated
with a definite
fi size of the alkyl substituent on the amide or thiourea group-
ing, with four to six carbons producing the most potent inhibitors. In spite
of their good antiChE properties, most of these compounds were not highly
effective as housefly
fl insecticides.
A substantial number of nitrophenyl methylcarbamates and dimethyl-
carbamates were examined for pesticidal activity; none were outstanding
either as an antiChE agent or as a toxicant (Fukuto et al. 1967). The p-
nitrophenyl carbamates were not as toxic to insects as the p-nitrophenyl
dialkylphosphates.

Multiple ring carbamates

The structure–activity relationships of multiring carbamates are of partic-
ular interest because of the signifificance of 1-naphthyl methylcarbamate
 Parameters for Carbamate Models 59

(carbaryl) as an insecticide. Double-ring carbamates are merely exten-

sions of doubly substituted phenyl methylcarbamates. Numerous other
2,3-ring additions exist, such as carbofuran, 2,3-dihydro-2,2-dimethyl-7-
benzofuranyl methylcarbamate. This compound represents a spatial ana-
logue of o-isopropoxyphenyl methylcarbamate (propoxur). Carbofuran is
one of the most effective of all the carbamate insecticides and has one of
the lowest LD50 values in the rat. The 3-OH and 3-C =O derivatives of
carbofuran show greatly reduced affinity
fi toward fly ChE, whereas the 3-
COCH3, 4-Cl, and 4-CH3 derivatives are essentially unaffected (Metcalf
et al. 1968). Substitution of 4- and 5-OH, on either the homonuclear ring
or the heteronuclear ring of carbaryl, has little effect on the affi
finity for
AChE, but it generally leads to a substantial reduction in toxicity for
insects (Metcalf 1971).

N-acylated carbamates
Fraser et al. (1968) substituted various acyl groups for the proton on the
N-methylcarbamates, producing N
N N-acyl carbamates that retained high
insecticidal activity and substantially reduced mammalian toxicity. Metabo-
lism is believed to involve deacylation to the parent carbamate with a cor-
responding increase in antiChE activity (Fahmy et al. 1966). In general, the
N-acyl derivatives are generally quite biologically unstable, although Sulli-
N
van et al. (1967) acetylated carbaryl as a means of increasing its stability
during gas chromatography. Acetylation is believed to aid penetration to
the site of action in the insect, where hydrolysis to the N-methylcarbamate
N
releases the parent carbamate. No information was available concerning
the metabolism of N-acyl-
N N-methylcarbamates in mammals. A reduction
N
in toxicity may well be related to aromatic ring hydroxylation or hydrolysis
of the carbamate over that of the acyl group.

Oxime carbamates
The structure–activity relationships of the N
N-methylcarbamoyl oximes were
reviewed by Felton (1968), Fukuto et al. (1969), and Payne et al. (1966).
The structural rigidity associated with the C=N bond is a unique feature
of the oxime carbamates. These carbamates form pairs of syn (Z isomer)
and anti (E isomer) isomers in which the oximino oxygen is either cis or
trans to the aldehyde H. The syn isomer of methomyl has 100 times the
antiChE activity and is 10 times more toxic to the housefl fly than the anti
isomer. Aldicarb is believed to exist as the syn isomer (Weiden 1968), and
it appears that this cis confi
figuration allows greater access of the carbonyl
carbon to the esteratic site and the methylthio group to the anionic site.
Methomyl is applied as a foliar spray, whereas the more toxic and environ-
mentally stable oxime carbamate, aldicarb, is applied to soil as a plant
systemic.
60 J.B. Knaak et al.

B. Carbamates and Their Toxicity

Carbamate pesticides belong to four chemical classes: oxime-N- N
methylcarbamates, aryl NN-methyl carbamates, N N-phenyl carbamates, and
methyl esters of substituted carbamic acids. Table 2 lists the common
name, chemical class, nominal mass, and CAS number of the 10 carbamates
of interest. Appendix A gives the chemical structures, physical parameters,
and tissue:partition coeffi
ficients of parent carbamates and metabolites.
Appendix B gives the metabolic pathways and preliminary metabolic rate
constants (Vmax, Km) for the metabolism of parent carbamates and
metabolites.
The toxicological data required by government agencies (e.g., USEPA
and California Department of Pesticide Regulation) to register carbamate
and other pesticides are extensive. A review of the registration require-
ments under the Federal Insecticide, Fungicide and Rodenticide Act
(FIFRA) and California Department of Food and Agriculture (CDFA)
regulations was made by Knaak et al. (1993). Acute toxicity studies provide
information needed to run PBPK/PD models and carry out the required
studies under FIFRA. Animal studies involving metabolism and ChE
inhibition are considered under Special Toxicology in the FIFRA data
requirements.

Acute oral toxicity

The acute oral toxicity of the 10 carbamates was taken from a number of
sources, as indicated in Table 3. The most toxic carbamate is aldicarb, fol-
lowed by carbofuran. Modelers should be aware of the LD50 of the carba-
mates and not exceed them in a modeling exercise. Enslein (1988) and
Enslein et al. (1998) developed TOPKAT, a QSAR program for predicting

Table 2. General Information on the Carbamate Pesticides.

N
N-Methyl Tables (in
carbamate Molecular Appendices
Common name class weight CAS no. A and B)

Aldicarb Oxime 190.26 116-06-3 14, 24

Carbaryl Aryl 201.20 63-25-2 15, 25
Carbofuran Aryl 221.25 1563-66-2 16, 26
Formetanate Aryl 221.26 22259-30-9 17, 27
Methiocarb Aryl 225.31 2032-65-7 18, 28
Methomyl Oxime 162.21 16752-77-5 19, 29
Oxamyl Oxime 219.26 23135-22-0 20, 30
Pirimicarb Aryl 238.29 23103-98-2 21, 31
Propoxur Aryl 209.24 114-26-1 22, 32
Thiodicarb Oxime 354.47 59669-26-0 23, 33
 Parameters for Carbamate Models 61

Table 3. Acute Oral Toxicity (LD50) of Carbamate Pesticides.

Rat, male Rat, female Mouse, male Mouse, female

Carbamate mg kg−1 bw mg kg−1 bw mg kg−1 bw mg kg−1 bw

Aldicarba 0.65j 0.3

Carbarylb 230j 128
Carbofurana 5.0 2.0k
6.5j
Formetanatec 21j 18
Methomyld 17j 24 10
Methiocarbe 14.0 16.0
10.0j
Oxamylf 3.1 2.5 3.3 2.3
2.5j
Pirimicarbg 33.1, 147 68 to 221 107
100j
Propoxurh 95 to 104 86
70k
Thiodicarbi 50 to100
160k
a
Lewis (1996).
b
Matsumura (1985).
c
Worthing (l979).
d
Tomlin (1994).
e
USEPA (1994) RED, Methiocarb.
f
Kennedy (1986).
g
Gagua (1976); Hayes and Laws (1991).
h
Verschueren (1983).
i
WHO/FAO (2000).
j
TOPKAT, Accelrys Software Inc., San Diego, CA.
k
Fahmy et al. (1970).

the toxicity of chemicals including those of the carbamates (e.g., in Table

3). The acute oral LD50 values listed in Table 3 are in the TOPKAT training
set. The LD50 model in TOPKAT consists of several class-specific fi models,
with the class determined by structures, not use. There are 12–15 separate
LD50 models. The training set has about 2,000 compounds distributed across
the models. The toxicity data were obtained from RTECS (Registry of
Toxic Effects of Chemical Substances, NIOSH, CDC, Atlanta, GA). The
limitations of the models are primarily based on the quality of the data in
RTECS. TOPKAT predicts the toxicity of compounds not in the training
set and indicates when predicted values lie outside the training set.

Acute dermal toxicity

Myers and DePass (1993) reviewed acute toxicity testing by the dermal
route. Regulatory guidelines recommend the rabbit for use in the acute
62 J.B. Knaak et al.

Table 4. Acute Dermal Toxicity (LD50) of Carbamate Pesticides.

Rat, male, Rat, female, Rabbit, male,

Carbamate mg kg−1 bw mg kg−1 bw mg kg−1 bw

Aldicarba 2.5 >5.0

Carbarylb 4,000 2,000
Carbofuranc 10,200
Formetanated
Methomyle >1,000 >5,000
Methiocarbf >5,000
Oxamylg >1,200 740
Pirimicarbh >500 >500
Propoxuri >1,000
Thiodicarbj >2,000
a
Lewis (1996); Hansch and Leo (1979).
b
Lewis (1996).
c
Crop Protection Handbook (2007).
d
No data.
e
WHO (1996).
f
Tomlin (1994).
g
Kennedy (1986).
h
Hayes and Laws (1991).
i
Verschueren (1983).
j
WHO/FAO (2000).

dermal toxicity test because of its sensitivity to dermally applied chemicals,

large treatment area, docility, availability, and large database. The rat or
guinea pig is often used for dermal studies when minimal chemical or
funding is available. Toxicity is often sex dependent, with the LD50 values
being greater for male than for female rats and rabbits (Myers and DePass
1993). Permeability constants are often two to three times greater for
females than they are for males, the difference being due in part to the
thickness of skin and the presence or absence of oils and waxes. The EPA
requires animal studies under FIFRA guidelines (Knaak et al. 1993). The
toxicity data in Table 4 were taken from a variety of sources. The Crop
Protection Handbook (2007) probably contains the most complete list of
carbamates used in agriculture along with their dermal toxicities. Aldicarb
is the most dermally toxic carbamate of the carbamates considered here.
Modelers should be aware of their dermal toxicity when building and
running carbamate PBPK/PD models.

III. QSAR Models for Predicting Biological Parameters

Used in PBPK/PD Models
Hansch et al. (2002) published an approach for organizing data on chemi-
cal-chemical and chemical-biological reactions. The importance of being
able to relate structure to biological activity is obvious when one considers
 Parameters for Carbamate Models 63

the large number of biological parameters (absorption/distribution, parti-

tion coeffificients, metabolic rate constants, and biological response) that are
needed in the construction of PBPK/PD models (see Table 1). The develop-
ment of these models depends largely on the availability of biological data
in the literature or on the development of good quality data in laboratory
studies. The cost of the latter appears to be prohibitive in current drug or
pesticide development programs. Sixty-one QSAR programs in the Hansch
et al. (2002) database involving oxidoreductases of all types were found to
focus on cytochrome P450 isozymes (CYPs) (e.g., algicides: Schmitt et al.
2000; antimalarials: Sinha et al. 1999).
Several structure–bioavailability relationship models have been pro-
posed in the last few years (Clark and Pickett 2000; Ekins and Rose 2002;
Klopman et al. 2000). The best known study is the “rule-of-five”
fi developed
by Lipinski et al. (2001) based on four molecular descriptors. The rule
generates an alert about absorption problems if two of the following condi-
tions are met: (1) molecular weight >500; (2) number of hydrogen-bonds
acceptors >10; (3) number of hydrogen-bond donors >5; (4) calculated log
P > 5.0; and (5) poor water solubility. Lipinski specifically
fi states that the
rule-of-fifive only holds for compounds that are not substrates for active
transporters (Lipinski 2000; Lipinski et al. 2001). When the rule-of-five
fi was
developed, information about drug transporters was very limited (Wu and
Benet 2005). Amidon et al. (1995) devised a Biopharmaceutical Classifica- fi
tion System (BCS) that categorized drugs into four classes according to
aqueous solubility and gastrointestinal permeability. A drug is considered
to be “highly soluble” when the highest dose is soluble in 250 mL water
over a pH range 1–7.5 at 37°C, whereas a drug is considered to be “highly
permeable” when absorption of parent and metabolites is >90% of an
administered dose. Wu and Benet (2005) suggested replacing permeability
in the classifification system with major routes of elimination. The change
will facilitate predictions, expand the number of Class 1 drugs eligible for
waiver of in vivo bioequivalence studies, and provide new insight into drug
elimination. Class 1 drugs have high solubility, high membrane permeabil-
ity, and are extensively metabolized (Wu and Benet 2005).

A. Human Oral Bioavailability

Yoshida and Topliss (2000) developed a QSAR model for predicting drug
oral bioavailability using the quantitative structure–bioavailability relation-
ship of 232 structurally diverse drugs. Oral bioavailability was assigned one
of four ratings and was analyzed in relation to physicochemical and struc-
tural factors by the ordered multicategorical classifi fication method using the
simplex technique (ORMUCS) method. The bioavailability classifications fi
are shown in Table 5.
Primary structural descriptors for drugs involved the distribution coeffi- fi
cient, log DpH 6.5, where 6.5 is the pH of the small intestine. The observation
that bioavailability generally followed acids > neutrals > bases led to the
64 J.B. Knaak et al.

a
Table 5. Bioavailability Classification.
fi

Class 1 Class 2 Class 3 Class 4

Rating 1 2 3 4
Bioavailability (%) ≤20 20–49 50–79 >80
a
Bioavailability Classification
fi System (Yoshida and Topliss 2000).

formulation of a new descriptor, Δ log D (i.e., log DpH 6.5 − log DpH 7.4). The
addition of 15 structural descriptors relating to well-known metabolic trans-
formations yielded a satisfactory QSAR equation, S(X) = Σwisi with Si, the
descriptor values, and Wi, the weighting factors. The optimum value for
log DpH 6.5 is −0.3, and a progressive negative impact on bioavailability
occurs when values move away from this level. A log DpH 6.5 value of 3.0
results in a reduction of the bioavailability score by 0.5 and a value of 5.0
by 1.3. This approach allowed the authors to obtain a correct classification
fi
rate of 71% for the training set and 60% for the 40 compounds included in
the validation set. Pintore et al. (2003a) obtained the data set from Yoshida
and Topliss (2000) and added 235 molecules. A large set of molecular
descriptors was examined, and the most relevant parameters were selected
using a procedure called genetic algorithm concepts and a stepwise tech-
nique (GA/SW). The aim of the project was to apply adaptive fuzzy parti-
tioning (AFP) to two bioavailability data sets. The AFP model improved
the power prediction about 10%–15% on the training and validation set
molecules. Pintore et al. (2003a) did not strongly support the use of the
physicochemical descriptors Δ log D and log P as they used topological and
electrotopological descriptors instead. Neither article discussed the nature
of the drug formulations used in the human bioavailability studies.
Carbamates are considered to be readily absorbed when dissolved in
corn oil, PEG 400, propylene glycol, or other relatively nontoxic solvents.
No information on the rate of absorption of these carbamates was found
in the literature. Model optimization procedures were used in a carbofuran
PBPK/PD models to simulate oral absorption (Zhang et al. 2006).

B. Toxicity Models
Venkatapathy et al. (2004) assessed the oral rat chronic lowest observed
adverse effect level (LOAEL) model in TOPKAT (Enslein et al. 1998).
The percentages of accurate predictions for nondatabase chemicals did not
compare well with predictions for chemicals in TOPKAT’s database. The
National Center for Environmental Assessment (NCEA), EPA, Cincinnati,
OH, used TOPKAT to screen/rank chemicals on the basis of their
hazard potential. The modules that are used with the greatest frequency by
NCEA are the rat chronic LOAEL, oral rat lethal dose (LD50), rodent
 Parameters for Carbamate Models 65

carcinogenicity, Ames mutagenicity, and developmental toxicity models.

TOPKAT has a built-in quality control module that determines if a chemi-
cal’s descriptors were within the model’s prediction domain. TOPKAT also
has a similarity search feature capable of determining whether a queried
chemical was represented in TOPKAT’s database.
In addition to TOPKAT, commercial QSAR software packages such as
DEREK (LHASA, Leeds, UK), CASE and MULTICASE (Multicase,
Cleveland, OH), COMPACT, HazardExpert (Compudrug, Budapest,
Hungary), and ONCOLOGIC (Logichem, Boyertown, PA) are able to
predict toxicity potentials for a wide variety of chemicals (Venkatapathy
et al. 2004).
Pintore et al. (2003b) used AFP to predict the toxicity of pesticides. Data
sets of 235 pesticide compounds, divided into three classes according to
their toxicity toward rats, were used. Carbaryl, carbofuran, methiocarb,
methomyl, oxamyl, pirimicarb, propoxur, and thiodicarb were included in
the training sets.
Goldblum et al. (1981) used the QSAR Eq. 1 to describe the relationship
between inhibition for 269 phenyl N N-methylcarbamates inhibiting fly head
AChE and the molecular descriptors MR (molar refractivity), Es (Taft
steric parameter), ᑣ (an inductive parameter), σ (Hammett parameter,
meta substituents), RGMR (MR of certain parts of ring substituents), CHG
(indicator variable for charged substituents), and HB (hydrogen bonding)
of the pesticides.
pI 0.557( 0.08)MR , , 1.558( 0.020)MR 0.611( 0.09)Es
− 0. ( . )(∑ ) . ( . )CHG
− 0.227( 0.04)MR 2 5. ( . )ᑣ , 2
2
. ( . )ᑣ2,6
+ 0.. ( . ) . (±0.22)HB
− 0.052(±0.02) 3 − 0.563( ±0.29)
2
2 × 6 + 3.458( ±0.21) (1)
where n = 269, r = 0.796, s = 0.485, Ideal ᑣ2,6 = 0.331(0.295 − 0.368), and
2

Ideal MR2 = 3.34(3.16 − 3.79).

Using the best correlation equation, the authors were able to predict the
concentration producing 50% inhibition for 269 different molecules within
a factor of ±3 (i.e., antilog of 0.485) for a concentration range of 1 million.
According to O’Brien (1960), it is more desirable to have ki values (bimo-
lecular rate of inhibition) for direct reference to the mechanism, since ki
values are directly related to the binding step:
k +1 k k
EH1 CB ← EHCB EC EH 2 C (2)
k −1

where EH1 = free enzyme, CB = carbamate inhibitor, EHCB = enzyme-

carbamate complex, EC = carbamylated enzyme, EH2 = regenerated
enzyme, and C = carbamic acid.
Kd = k−1/k+1 (equilibrium or affi
finity constant) (3)
66 J.B. Knaak et al.

ki = k2/Kd (bimolecular inhibition rate constant) (4)

Abd-Elroaf et al. (1977) and Nishioka et al. (1977) found k2 to be more
or less constant for a set of carbamates. The long reaction periods used in
the I50 studies (Kolbezen et al. 1954) complicated the otherwise relatively
straightforward relation of ki to I50 (O’Brien 1960) because the decarba-
mylation rate constant, k3, has a shorter half-life, t1/2, than the span of time
used for the I50 measurements. The parameter ki turns out to be insensitive
to the concentration of the inhibitor used (Nishioka et al. 1976) in the
kinetic runs, so it is of greater comparative value for results from different
sources. Nevertheless, the authors believe that data with a large set of
congeners can be of real value in enabling them to rationalize some of the
salient events of the binding step and in exploring the characteristics of the
binding site via physicochemical correlation studies.

C. Liver Microsomal P450 and CYP Hydroxylation Models

QSAR models were developed by Enslein et al. (2007) for predicting Vmax
and Km phase I reactions involving the hydroxylation of (1) drug aromatic
and (2) alicyclic rings and aliphatic groups by human liver microsomes
(HLM) and by CYP3A4 isozyme. Additional drug-based QSAR-Vmax, Km
models involving CYP1A2, 2D6, and 2C9, individually, are under develop-
ment. Approximately 50% of the biotransformations involving drugs are
hydroxylation reactions, the remaining transformations include N- and O-
demethylations. Hydroxylated drugs are conjugated with glucuronic or sul-
furic acid in phase II reactions. A number of carbamates are either
ring-hydroxylated or their N N-methyl groups are hydroxylated. Carbaryl
(Tang et al. 2002) is converted to 4-OH, 5-OH, and hydroxymethyl deriva-
tives, whereas carbofuran (Usmani et al. 2004a) is converted to 3-OH car-
bofuran. Vmax, Km data (e.g., Vmax, nmol min−1nmol−1 CYP3A4; Km, μmol L−1),
from these two carbamates were used in the training sets for the CYP3A4
hydroxylation model along with data from 66 drugs. HLM QSAR − Vmax, Km
models (e.g., Vmax, μmol hr−1 kg−1 bw; Km, μmol L−1) were developed without
carbamate data. The CYP3A4 models turned out to be stable and useful for
predicting Vmax, Km values, while the HLM models were not stable and could
not be used. The reason(s) for the instability appears to be associated with
variation in CYP3A4 content in the microsomes.

D. Tissue:Blood Partition Coefficients

fi
Zhang (2005) developed a nonlinear QSAR equation using (1) published
tissue:blood partition coeffi ficients (PCts) of 36 organic solvents in human
fat, liver, brain, kidney, muscle, lung, and heart, and (2) PCts of 10 drugs
for rabbit fat, brain, muscle, lung, and heart. No pesticidal carbamates were
 Parameters for Carbamate Models 67

included in the training sets. The model takes into consideration the volume
of the tissue composition (lipid, protein, and water), and the form(s) of the
compound (cationic, anionic and neutral) at pH 7.4. The present research
only included neutral and cationic forms. The QSAR equation took the
following form:

log t logg(∑ ) (5)

where subscript i = l, p, w (indicate the lipid, protein, and water in tissue,

respectively); and subscript j = ui, +, − (indicate the neutral form, cationic
form, and anionic form of a compound, respectively).
The author used the following molecular descriptors in the QSAR
model: molecular polarizability (α), maximum positive charge (q+max), sum
of all positive partial atomic charges for all atoms in the molecule (ΣQ+),
the sum of H-bond factor values for all acceptor substructures in the mol-
ecule (ΣCa), the sum of H-bond factor values for all donor atoms in a
molecule (ΣCd) and the maximum H-bond acceptor descriptor in a mole-
cule (Camax). All descriptors were calculated by the program package
HYBOT/HYBOT-PLUS-98 (Raevsky 1997). Nonlinear regression analy-
ses were performed using a standard regression program (GFA BASIC
4.38; GFA Software Technologies, Moenchengladbach, Germany). The
descriptors were introduced into Equation 5 in a stepwise manner until the
statistical results could not be further improved. Equation 5 states that the
PCt may be expressed as the weighted sum of three PCts (lipid:blood,
protein:blood, and water:blood) of neutral and ionic forms. The equilib-
rium distribution of a chemical is essentially the sum of the distributions of
the various chemical forms. Five parameters were used to fit fi neutral com-
pound partitioning into tissues, and two parameters were used for ionic
compounds. The partitioning of ionic compounds into lipids increases with
increasing Camax. The q+max value plays an important role in compounds
partitioning into protein. The model equation is the expressive form of the
Hansch equation (Hansch et al. 1962) in nonlinear spaces (multiphase
system).
Liu et al. (2005) combined information on the tissue:blood partition
coefficients
fi of 35 organic compounds in seven different tissues with the
lipid, protein, and water content of the seven tissues with their molecular
descriptors to select the best descriptors for a QSAR model. The model
represented one tissue composition descriptor, two electrostatic descrip-
tors, one quantum chemical descriptor, and one geometric descriptor. The
weight fraction of water (WF) in different tissues was the most important
descriptor affecting a tissue:blood partition coeffi ficient. A least squares-
support vector machine (LS-SVM) was used to develop the nonlinear
QSAR model. All calculations of the LS-SVM were performed using the
MATLAB/C toolbox. No carbamates were present in the data set.
68 J.B. Knaak et al.

IV. Experimentally Derived Biological Parameters Used in

PBPK/PD Models
A. Gastrointestinal Absorption
Bolus dose
Timchalk et al. (2002) used absorption rates of 0.01, 0.5, and 0.5 hr −1
for stomach to blood, intestine to blood, and stomach to intestine for chlor-
pyrifos (CAS no. 2921-88-2). A bolus dose (chemical in mg kg−1 bw or
μmol kg−1 bw) is placed in the stomach at zero time. A number of fifirst-order
rate constants describe the absorption of the chemical from the stomach
and intestine to blood. A first-order rate constant is also used to transfer
chemical from stomach to intestine. A bioavailability factor of 0.8 (class 4)
from Table 5 reduces the amount of the dose.

Rate of ingestion
In the case of a rate ingestion using the Exposure Related Dose Estimating
Model (ERDEM) (USEPA 2004), a dose (concentration in dosing liquid,
mg mL−1 or μmol mL−1) is administered at a uniform fl flow rate (mL min−1)
over a period of time (min) to give a dose in (mg or μmol) in the stomach.
The dose (mg), as it arrives in the stomach, is then absorbed as indicated
for a bolus dose. Total dose (mg) is calculated as follows: mg mL−1 × mL min−
1
× min. In practice, if the time of administration is rapid, the dose acts like
a single bolus dose.

B. Skin Absorption
History
Illnesses associated with the use of ChE-inhibiting pesticides (i.e., OP com-
pounds and N N-methylcarbamates) have generated considerable field
research into the relationship between ChE inhibition and residues in air,
on treated indoor surfaces, and on crop leaf surfaces (Honeycutt and Day
2001; Honeycutt et al. 1985; Siewierski 1984; Wang et al. 1989). This work
was accompanied by the development of information pertaining to the
science of percutaneous absorption (Bronaugh and Maibach 1985; Scott
et al. 1990) and the linking of field
fi exposures to percutaneous absorption
and ChE inhibition (Knaak et al. 1989, 2002; Nigg and Knaak 2000; USEPA
1992). The information in these studies was used to develop code within
ERDEM to predict the effects of dermal exposure to ChE-inhibiting pes-
ticides on ChE activity in exposed individuals. Knaak et al. (2002) described
the relationship between skin permeability, Kp (cm hr−1), and partition coef-
ficients of skin:air, skin:vehicle, and skin:blood. In the case of skin:air, a
fi
Kp-vapor value (cm hr−1) takes into consideration the partitioning of vapor
between air and skin, leading to a concentration in the skin and a percu-
taneous absorption rate (μg hr−1 cm−2). The permeability constants for
 Parameters for Carbamate Models 69

pesticides in delivery solvents, Kp-solvent, are not determined in most studies.

Skin absorption rates are reported in μg hr−1 cm−2. The solvent, xylene, has
been shown to enhance the penetration of methyl parathion over that of
acetone in studies involving cadaver skin. Acetone has been used exten-
sively for applying a pesticide on skin. The solvent rapidly evaporates,
leaving the pesticide to be absorbed by skin or to evaporate into the air
(Bronaugh and Maibach 1985).

Relationship between Kp (permeation constant), skin:air, and

skin:vehicle partition coefficients
fi
A direct relationship exists between the permeation constant, Kp (cm hr −1),
ficient D (cm2 hr−1), the thickness of skin l (cm), the con-
the diffusion coeffi
centration C of the pesticide in skin (g cm−3), and the skin:vehicle partition
coefficient
fi km (unitless) based on the following equation from Mattie et al.
(1994).
Flux = DkmC/l = KpC (6)
In the case of pesticides in vehicles such as water or air, skin:air and skin:
water partition coefficients
fi need to be determined and used in the equation
to account for higher concentrations in skin caused by partitioning.

Skin:blood partition coefficients

fi
Skin:blood partition coefficients
fi are used in PBPK/PD models to transfer
pesticides between skin and blood. Jepson et al. (1994) developed a labora-
tory procedure for measuring the partitioning of chemicals between skin
and blood using low-binding Millipore cellulose filters,
fi and Poulin and
Theil (2002a) developed a mechanistic method for measuring tissue:blood
partition coefficients.
fi The procedures are discussed in detail in Section
IV.C.

Dose–response, pharmacokinetic, and metabolism studies involving

carbamate pesticides
Knaak et al. (1984) and Knaak and Wilson (1985) developed 7-d pharmaco-
kinetic data on the percutaneous absorption of carbaryl (43.5 μg cm−2,
13.8 cm2 skin) through the intact back skin of the rat using 14C-naphthyl car-
baryl. The skin permeability constant ranged from 4.0 × 10−3 to 7.1 × 10−3
(cm hr−1) based, respectively, on the t1/2 values for elimination from
plasma or loss from skin. Skin absorption rates varied between 0.18 and
0.31 μg cm−2 hr −1 (i.e., Kp = 0.31/43.5 μg cm−2 hr−1 = 7.1 × 10−3 cm−2 hr−1). No
information was obtained by Knaak et al. (1984) concerning the nature of
metabolites in skin or eliminated in urine. The fate of 14C naphthyl carbaryl
in rat skin was studied by MacPherson et al. (1991) in the presence or
absence of NADPH (nicotinamide adenine dinucleotide phosphate,
70 J.B. Knaak et al.

reduced), UDPGA (uridine diphosphate glucuronic acid), or PAPS (3′-

phosphoadenosine 5′-phosphosulfate). 1-Naphthol was formed by postmi-
tochondrial fractions from the liver and skin with and without cofactors. In
the presence of NADPH, hydroxycarbaryl was formed by the liver but not
by skin postmitochondrial fractions. 1-Naphthyl glucuronide and sulfate
were formed by postmitochondrial fractions from the liver and skin. No evi-
dence of carbaryl metabolism was detected in studies involving intact skin.
In a pharmacokinetic percutaneous study with 14C-labeled thiodicarb,
thiodicarb was poorly absorbed from skin, with 4.4% found in urine, 2.5%
in feces, 8.8% in the carcass, and 6.6% in respiratory air CO2 and acetoni-
trile (Knaak and Wilson 1985). The remainder of the dose was found on
the surface of the skin. Thiodicarb was absorbed during the first fi 24 hr at a
rate of 0.27 μg cm−2 hr−1 (Kp = 0.27/48 = 5.6 × 10−3 cm hr−1). The rate was
similar to that of carbaryl. After 24 hr, the rate of loss from skin decreased
to 0.042 μg cm−2 hr−1 (Kp = 0.042/48 = 8.75 × 10−4 cm hr−1), and remained so
until the study was terminated at 168 hr. The reason for the rate change
appears to be associated with the physical properties of the applied thiodi-
carb, and its distribution across treated skin. Microscopic examination of
treated skin revealed crystals of thiodicarb deposited on hair and on the
surface of the skin suggesting that only a portion of the applied dose was
readily available for absorption when using acetone as a vehicle. Vehicles
such as alcohol have a tendency to increase percutaneous absorption.
Van de Sandt et al. (1993) topically applied 14C propoxur at various
concentrations to human, rabbit and porcine skin disks in vitro. Permeation
rates in human skin were 9.2, 40.7, and 56.6 ng cm−2 hr−1, respectively, for
applied dosages of 25, 100, and 200 μg cm−2. The experimental period
was 6 hr. The metabolites in skin (1.1%–4.9%) were identified fi as IPP (2-
isopropoxyphenol), IPP-glucuronide, IPP-sulfate, and an unknown metab-
olite. Human skin produced only sulfate conjugates, rabbit skin produced
glucuronides, and porcine skin produced sulfates and glucuronides. Com-
parative in vitro-in vivo percutaneous absorption studies involving pro-
poxur were carried out by Van de Sandt et al. (2000). For direct comparison,
experimental conditions involving dose (150 μg propoxur cm−2), vehicle
(60% ethanol), and exposure time (4 hr) were standardized. Percutaneous
penetration in human volunteers was measured by the analysis of 2-
isopropoxyphenol in blood and urine. Maximal fl flux (μg cm−2 hr−1) was cal-
culated from the linear portion of the cumulative penetration curve. The
permeability coefficient
fi (Kp value in cm hr−1) [H3] H2O was calculated
according to ECETOC (1993), by dividing the maximal flux (μg cm−2 hr−1)
by the applied dose. In the in vivo rat studies, propoxur was applied to an
area of 10 cm2 (total dose, 1,500 μg), while in the human studies, an area of
100 cm2 was utilized on the forearm (total dose, 15,000 μg). Maximal fl flux in
the rat and human were 8.7 and 1.0 μg cm−2 hr−1, respectively. Kp values were
estimated to be 8.7/150 = 5.8 × 10−2, and 1.0/150 = 6.6 × 10−3 cm hr−1, respec-
tively, for the rat and human.
 Parameters for Carbamate Models 71

Additional pharmacokinetic studies involving pesticidal carbamates

were not found in the published literature. The results of the studies involv-
ing carbaryl, thiodicarb, and propoxur suggest that carbamates are not as
readily absorbed through skin as OP pesticides.

C. Tissue Partition Coefficients/Distribution

fi
Partition coefficients
fi from in vitro data
Tissue:blood partition coeffi ficients are required for the development of
PBPK models. Jepson et al. (1992, 1994), Knaak et al. (1995), and Kousba
and Sultatos (2000) are examples of this work involving the use of nonvola-
tile chemicals. According to the procedure of Jepson et al. (1992), Knaak
et al. (1995) used heparinized rat blood, homogenized rat tissues consisting
of liver, kidney, muscle (thigh), fat (perirenal), and brain. The tissues
(0.32 g) suspended in 20 mL 0.9% saline in triplicate vials (test and refer-
ence) were placed in a shaking water bath at 37°C to allow the contents to
come to thermal equilibrium. Reference vials contained 20 mL 0.9% saline.
Isofenphos (150 μg in 50 μL ethanol) was added and contents equilibrated
for 24 hr in 20 mL 0.9% saline. A 2-mL aliquot was ultrafiltered,
fi and 100-μL
aliquots of the filtrate were analyzed for pesticide. Equation 7 was used to
determine the amount of pesticide recovered in the reference saline after
ultrafifiltration.

Rf = Csal /Cfil (7)

where Rf is the recovered fraction, Cfil is the concentration of the pesticide

in the ultrafi
filtrate from reference saline, and Csal is the concentration of the
pesticide in the reference saline before ultrafiltration.
fi
The concentration in tissue or blood was calculated from the concentra-
tion found in the sample ultrafi filtrate using Eqs. 8 and 9.

Assal = Css(Vss)/Rf (8)

Cj = (Arsal − Assal)/V
Vj (9)

where Assal = the amount of pesticide in the sample saline after filtration,
fi
Css = the concentration of pesticide in the sample saline after fifiltration,
Vss = the volume of saline in the test vial,
Arsal = the amount of pesticide in the reference saline,
Vj = the volume of tissue, and
Cj = the concentration of pesticide in tissue j.
The tissue : saline and tissue : blood PCts are calculated using Eq. 10.

Pj = Cj(V
Vj)/Assal (10)

where Pj is the partition coeffi

ficient for tissue j (blood, liver, etc.) and Assal,
Vj, and Cj are described above.
72 J.B. Knaak et al.

Several diffificulties are encountered in carrying out this procedure: (1)

nonspecific
fi binding sites on filters and filtration apparatus often retain
pesticides; (2) filtration rates are slow even under pressure; and (3) enzyme
inhibitors should be used to prevent metabolism. The PCts are then calcu-
lated by dividing the tissue : saline partition coefficients
fi by the blood : saline
partition coefficients.
fi Isofenphos rat tissue partition coeffi ficients were 48,
21, 17, 27, and 23, respectively, for fat, liver, brain, kidney and muscle
(Knaak et al. 1995). The values obtained with the Poulin and Theil (2002a)
mechanistic method described later in this review were 358, 11.6, 28.1, and
6.49, respectively, for fat, liver, brain, kidney, and muscle.
Jepson et al. (1994) modifiedfi their earlier 1992 method by changing tissue
weights (1.0 g fat, 1.5 g muscle, 0.25 g liver, and 0.5 g blood) and adding
centrifugation before ultrafiltration.
fi Ultrafifilration was carried out using a
Millipore Low Binding Celluose Membrane Filter Unit with a 10,000
nominal molecular weight (Millipore Ultrafree-PF; Millipore, Bedford,
MA). The membranes were attached to the bottom of a filter fi cup as part
of the manufacturing process. The system with filter cups is shown in Fig.
1. Each cup contained a 10-mm. four-pointed Teflon fl stir bar (Starburst;
Parkside Industries, Chicago, IL). The partition coefficients
fi for parathion
(CAS no. 56-38-2) were 103, 2.59, and 5.42, respectively, for fat, muscle,
and liver. The values using the Poulen and Theil (2002a) mechanistic method
were 295.9, 6.3, and 11.4, respectively, for fat, muscle, and liver.

Fig. 1. Schematic of the system used to filter samples containing chemical equili-
brated between tissue and saline. (Redrawn from Jepson et al. 1994 by permission
of Oxford University Press.)
 Parameters for Carbamate Models 73

In the two experimental partition coeffi ficient studies (Jepson et al. 1994;
Knaak et al. 1995) involving OPs, the fat: blood PCs were significantly fi
below those estimated by the Poulen and Theil (2000) method. An increase
in the amount of fat (0.32 to 1.0 g) used by Jepson et al. (1994) in their
parathion (log P, 3.84) studies had a positive effect on this PC over the value
obtained by Knaak et al. (1995) for isofenphos (log P, 4.12). The differences
between experimental PC values and those obtained using mechanistic
models needs to be carefully examined before a recommendation can be
made regarding the use of in vitro over mechanistic values.

Tissue : Blood Partition Coeffi

ficients from Mechanistic Models
QSAR models by Zhang (2005) and Liu et al. (2005) were described in
Section III. D: Tissue : Blood Partition Coefficients.
fi The models did not
contain carbamates or their metabolites in the training set and may not be
suitable for predicting tissue : blood partition coefficients
fi for carbamates.
The models look sound, and the procedures used in their development
should be pursued in the development of QSAR models for carbamates.
Mechanistic approaches for prediciting partition coefficientsfi from chem-
ical structure information (log P) and tissue : plasma composition were
developed and validated by Poulin and Krishnan (1996), Poulin and Theil
(2000), Poulin et al. (2001), Poulin and Theil (2002a), and Poulin and Theil
(2002b). Drug Po:w (n-octanol : buffer, nonionized species at pH 7.4), Dvo:w
and D*vo:w (vegetable oil : buffer or olive oil : buffer, ionized and nonionized
species at pH 7.4) were used in Eqs. 11 and 12 with tissue : plasma lipid and
water, and unbound content in tissue : plasma to calculate PCt:p_nonadipose and
PCt:p_adipose values (Poulin and Theil 2002a,b).

[Po w (Vnlt . Vpht

p )] [ ( Vwt . p
pht)] fu p
PC t p nonadipose = × (11)
[Po w ( p . p p )] [ ( p . p p )] fu t
[D*vo w or Dvo w (Vnlt . Vpht )] [ (Vwt . Vpht
h )] fu p
PC t p adipose = × (12)
[D*vo w or Dvo w (Vnlpp . Vpphpp )] [ (Vwpp . Vpphpp )] 1

where V is the fractional tissue volume content of neutral lipids (nl),

phospholipids (ph), water (w), tissue (t), and plasma (p), fut is the macro-
molecular unbound fraction in tissues, and fup is the macromolecular
unbound fraction in plasma. The ratio fup/fut is equal to Cm tissue/Cm plasma
[(C = concentration; m = main binding macromolecules present in these
two matrices, i.e., albumin, globulins and or lipoproteins (HDL, LDL)]
with the ratio varying from 0.3 to 1 in mammalian tissues. Consequently,
this contribution factor was not considered in determining PCt when pre-
dicted values differed from experimental values by a factor less than 2
(Poulin and Theil 2000). Plasma protein binding was considered as a sepa-
rate issue in a PBPK/PD model published by Timchalk et al. (2002) on
chlorpyrifos.
74 J.B. Knaak et al.

In other terms, in vivo PCt:p_adipose would only be dependent on the lipo-

philicity of nonionized species if the quantitative role of ionized species and
plasma protein binding is not taken into account. The neutral nonpolar
lipids of nonadipose tissues are mainly mixtures of triglycerides and cho-
lesterol, and n-octanol mimics the lipophilicity of this mixture. The adipose
tissue lipids are composed mostly of triglycerides, and olive oil is a good
surrogate solvent. For cases involving only nonionized species in the
aqueous phase, Dvo:w was obtained using the relationship between Po:w and
Dvo:w in Eq. 13, as developed by Leo et al. (1971).
log Dvo:w = 1.115 log Po:w − 1.35 (13)
with n = 104, r = 0.99.
Fouchecourt et al. (2001) used the KowWin program (Meylan and
Howard 1995) to obtain log Po:w for n-hexane, and the Leo et al. (1971)
equation for log Kvo:w.
log Kvo:w = 1.099 log Po:w − 1.31 (14)
Equation 14 is comparable to Eq. 13. For the prediction of Pt:p_adipose with
Eq. 12, data on D*o:w (nonionized and ionized species) are required instead
of Dvo:w (nonionized). Log D*o:w values were determined using Eq. 13 (i.e.,
log Dov:w) and the Henderson–Hasselbach equations for mono-, diprotic
acids, bases, and zwitterionic chemicals as follows:
Monoprotic acid:
log D*o:w = log Dvo:w − log(1 + 10pH−pKa 1) (15)
Monoprotic base:
log D*o:w = log Dvo:w − log(1 + 10pKa 2−pH) (16)
Diprotic acid:
log D*o:w = log Dvo:w − log(1 + 10pH−pKa 1+pH−pKa 2) (17)
Diprotic base:
log D*o:w = log Dvo:w − log(1 + 10pKa 1−pH+pKa 2−pH) (18)
Zwitterionic:
log D*o:w = log Dvo:w − log(1 + 10−pKa 2+pH+pKa 1−pH)
pKa1 = acid, pK Ka2 = base (19)
Note that Eqs. 15–19 require information on the pKa of the acid. Poulin
and Theil (2002a) assumed that D*o:w and Dvo:w differ by a factor corre-
sponding to the ionized species in the aqueous phase. According to Lewis
(2000), log D7.4 involving octanol : water is a combination of lipophicity
(the log P term) and ionization (the pK Ka term) with the relationship being
7.4
log Do:w = log Po:w − log(1 − 10pH−pKa ) for monoprotic acids. This equation is
the same as Eq. 15 except for the fact that distribution takes place between
vegetable oil : water in Eq. 15 and not octanol : water as described by Lewis
(2000).
 Parameters for Carbamate Models 75

The relationship between log Po:w and log Dvo:w for adipose tissue : plasma
and adipose tissue : blood is shown in Figs. 2 and 3. Data from Parham
et al. (1997) for polychlorinated biphenyls (PCBs) are included in the adi-
pose : plasma fifigure, and data from Jepson et al. (1994) are included in the
adipose : blood figure. Tissue : blood PCts are routinely used in PBPK
models. The use of log Dvo:w reduces the partition coeffi ficients by approxi-
mately a factor of 10; however, this factor is not greatly reduced for che-
micals with log Po:w greater than 3. All the PCBs had log Po:w partition
coefficients
fi greater than 4.0; this is also the case with most of the OPs, but
not with the carbamates.
The log Dvo:w model appears unbiased when log P = 0 to 2 and biased
when log P > 3. The experimental parathion log P values of Jepson et al.
(1994) for parathion are shown along with values predicted by the model.

Selection of log Po:w and log Do:w Values for Use in Mechanistic Models
The key to using the mechanistic approach is the selection of a log P/log D
model that adequately predicts values for the 10 carbamates and their
metabolites (ionized and nonionized species). Log P models include Clog P
(Leo 1993), KLog P (Klopman et al. 1994), HLog P (Viswanadhan et al.
2000), ALog P (Wildman and Crippen 1999), XLog P (Wang et al. 2000),
ACDLog P (Petrauskas and Kolovanov 2000), SMILELOG (Convard
et al. 1994), CHEMICALC (Suzuki and Kudo 1990), and LOGKOW

Fig. 2. Adipose : plasma partition coefficients

fi vs. log Po:ww (and log Dvo:w). 䉬 Log Po:w
(Knaak et al. 2005); ⵧ log Dvo:w (Leo et al. 1971); Δ PCB data (Parham et al.
1997).
76 J.B. Knaak et al.

Fig. 3. Adipose : blood partition coefficients

fi vs. log Po:w (and log Dvo:w). 䉬 Log Po:w
(Knaak et al. 2005); 䊏 log Dvo:w (Leo et al. 1971); 䊊 experimental log Po:w (Jepson
et al. 1994); Δ predicted log Po:w (Jepson et al. 1994). Points at the bottom are for
paraoxon; points at the top are for parathion.

(Meylan and Howard 2000). Hansch et al. (2002) currently have a database
containing almost 30,000 experimentally measured octanol/water log P and
log D values for constructing QSAR equations. The importance of log P
values cannot be overestimated, as Hansch et al. (2002) lists 4614 QSARs
with log P terms in a list of 8500 QSAR equations.
ACD Log D Sol Suite, Version 9.0 (Advanced Chemistry Development,
Toronto, Ontario, Canada) was selected for predicting pKa, log P, log D,
and water solubility (WS) values. The program is commercially available
and is used by the American Chemical Society to predict chemical and
physical properties of chemicals listed in SciFinder (American Chemical
Society, Washington, DC). Version 9.0 predicts log D at any pH and pro-
vides a printout of log D values in units of 0.1 pH.
As part of the selection process, the WS for each of the carbamates and
their metabolites were individually plotted against log DpH7.4 values to
determine whether a linear, but inverse, relationship existed based on the
knowledge that metabolism significantly
fi increases the water solubility of
administered pesticides, drugs, and other chemicals. Water solubility was
calculated using the equation of Meylan and Howard (1994a,b) where
Log S(mol/L) = 0.796 − 0.854 log Ko:w − 0.00728MW + corrections (20)
ACD log D values were used in the equation in place of log Ko:w to calculate
water solubilities. This procedure yielded linear plots with r 2 values greater
 Parameters for Carbamate Models 77

Fig. 4. Water solubility (g L−1, Eq. 20) of aldicarb and metabolites as a function of
ACD log DpH7.4. Aldicarb and metabolites are identifi fied in Table 14 (Appendix A).
Outer bounds represent the 95% confidence
fi interval.

than 0.9 for all 10 carbamates and their metabolites. Figures 4 and 5 show
the plots for aldicarb and carbaryl. Water solubilities >10,000 (g L−1) were
calculated by the Meylan and Howard (1994a,b) equation for conjugated
carbamate metabolites but were not provided by the ACD log D suite.
These results were not obtained using log Ko:w values (Discovery Studio
Accord for Excel; Accelrys, San Diego, CA) for carbamate metabolites
because log Ko:w alone did not adequately predict values for water-soluble
ionized metabolites.
78 J.B. Knaak et al.

Fig. 5. Water solubility (g L−1, Eq. 20) of carbaryl and metabolites as a function of
ACD Log DpH7.4. Carbaryl and metabolites are identifi fied in Table 15 (Appendix
A). Outer bounds represent the 95% confidencefi interval.

Partition Coeffi
ficients for Carbamates and Metabolites
On the basis of the foregoing findings
fi involving water solubility for aldicarb
acid, log DpH7.4 (−2.56), determined by the ACD Log D Suite was used
in conjunction with an Excel spreadsheet (Table 6) to calculate log Dvo:w
(−4.20) from log DpH7.4 (−2.56) (Eq. 13) with log D*o:w = log Dvo:w (−4.20)
(Eq. 15, no pK Ka correction factor required) to obtain the adipose tis-
sue : blood partition coeffi
ficients for aldicarb acid (see Table 14, Appendix
A) using Eq 22. In the case of nonadipose tissue, Eq. 21 was used. When
log P (0.87) was used in Eq. 13, a value of −0.38 was obtained for log Dvo:w
 Parameters for Carbamate Models 79

and a value of −4.10 with Eq. 15 for log D*o:w (pH, 7.4 and pKa, 3.68). The
results for log D*o:w compared favorably with the value obtained using
log DpH7.4.
In the case involving a glucuronide such as 4-OH carbaryl glucuronide,
log D*o:w (pH, 7.4 and pKa, 2.75) was −6.63 using log P (−0.57). When
log DpH7.4 (−4.28) was used the value for log D*o:w was −6.12. In the case of
a sulfate, such as 4-OH carbaryl sulfate, log D*o:w (pH, 7.4; pKa1, −4.58 and
pKa2, 11.64) was −6.17 using log P (1.57). When log DpH7.4 (−1.93) was used
the value was −3.50 for log D*o:w. The reason for the difference is related
to the manner in which ACD Log D Suite computes log DpH7.4 versus the
Henderson–Hasselbach equation (Eqs. 15 and 17) for a mono- or dipro-
tic acid. On the basis of the differences observed we decided to use
log DpH7.4 values for determining partition coefficients
fi for all carbamate
metabolites.
The tissue : plasma and tissue : blood partition coefficients
fi were deter-
mined using Eqs. 21 and 22 (see Table 6). Tissue : blood partition coeffi- fi
cients are depicted below.
[Do w or Po w (Vnlt . Vphlt
p )] [ (Vwt .7 Vpphlt )] fu b
PC t b nonadipose = ×
[Do w or Po w (Vnlb . Vphlp
p p )] [ (Vwb . Vpphlb )] fu
f t
(21)
[D*vo w or Dvo w (Vnlt . Vpht )] [ (Vwt .7Vpht
h )] fu b
PC t b adipose = × (22)
[D*vo w or Pvo w (Vnlb . Vphb
p )] [ (Vwb . Vpphb )] 1

where PCt:b_nonadipose and PCt:b_adipose = partition coefficient

fi between tissue and
blood, pH 7.4, distribution coeffi ficient (D), partition coeffi ficient (P), octa-
nol : water (o : w), vegetable oil : water (vo : w), V is the fractional tissue
volume content of neutral lipids (nl), phospholipids (ph), water (w), tissue
(t), and blood (b), fut is the macromolecular unbound fraction in tissues,
and fub is the macromolecular unbound fraction in blood.
The partition coefficients
fi for the 10 carbamates and their metabolites
in adipose tissue, brain, rapidly perfused tissue (e.g., heart), kidney,
liver, slowly perfused (e.g., muscle), and skin are shown in Tables 14–23
(Appendix A).

Graphs of log DpH 7.4 vs. Adipose Tissue : Blood and Liver : Blood
Partition Coefficients
fi
The plots of log Do:w (pH 7.4) vs. PCt:b_liver and log D*o:w (pH 7.4) vs.
PCt:b_adipose for aldicarb and carbaryl are shown in Figs. 6 and 7.
In Figs. 6 and 7, the water-soluble metabolites, with log DpH7.4 < 0, all
have PCt:b_liver and PCt:b_adipose values ≤1.0 at the bottom of each plot, while
the parent carbamates and their neutral metabolites, with log DpH7.4 > 0,
have PCt:b values ranging from 1.0 to 40.0 extending up and on the right
side of each plot.
Table 6. Example Spreadsheet Used in Calculating Partition Coefficients
fi (Eqs. 21, and 22) for aldicarb acid in Table 14 (Appendix A).
80

Input parameters

log DpH7.4 o:w −2.6

pHa 7.4
pK
Kaa 3.68
log D*vo:w −4.2
log Dvo:w −4.2
fup 1
fut 1
fup/fut 1

Lipids

Neutral Phospho

Rat tissue Eq. no.b Tissue (Vt)c Water (Vw)c (Vnl)c (Vpl)c Ko:wd Numeratore Pt:bf Pt:pg Vt*Pt:ph

Adipose 22 0.0761 0.12 0.853 0.002 6E-05 0.1215 0.14 0.13 0.01
Adipose 21 0.0761 0.12 0.853 0.002 0.0028
J.B. Knaak et al.

Bone 21 0.041476 0.446 0.0273 0.0027 0.0028 0.4480 0.53 0.47 0.02
Brain 21 0.0057 0.788 0.0392 0.0533 0.0028 0.8255 0.98 0.86 0.00
Gut 21 0.027 0.749 0.0292 0.0138 0.0028 0.7588 0.90 0.79 0.02
Heart 21 0.0033 0.779 0.014 0.0118 0.0028 0.7873 0.94 0.82 0.00
Kidney 21 0.0073 0.771 0.0123 0.0284 0.0028 0.7909 0.94 0.82 0.01
Liver 21 0.0366 0.705 0.0138 0.0303 0.0028 0.7263 0.86 0.76 0.03
Lung 21 0.005 0.79 0.0219 0.014 0.0028 0.7999 0.95 0.83 0.00
Muscle 21 0.404 0.756 0.01 0.009 0.0028 0.7623 0.91 0.79 0.32
Skin 21 0.19 0.651 0.0239 0.018 0.0028 0.6637 0.79 0.69 0.13
Spleen 21 0.002 0.771 0.0077 0.0136 0.0028 0.7806 0.93 0.81 0.00
Plasma 21 0.0449 0.96 0.00147 0.00083 0.0028 0.9606 0.631i
Plasma 22 0.0449 0.96 0.00147 0.00083 6E-05 0.9606h
Whole blood 21 0.0816 0.84 0.0013 0.002 0.0028 0.8414g
Whole blood 22 0.0816 0.84 0.0013 0.002 6E-05 0.8414f
Erythrocytes 21 0.0367 NUj NU NU
 Lipids

Neutral Phospho

Human tissue Eq. no.b Tissue (Vt)c Water (Vw)c (Vnl)c (Vpl)c Ko:wd Numeratore Pt:bk Pt:pl Vt*Pt:pm

Adipose 22 0.11957 0.18 0.79 0.002 6E-05 0.1814 0.22 0.19 0.02
Adipose 21 0.11957 0.18 0.79 0.002 0.0028
Bone 21 0.085629 0.439 0.074 0.0011 0.0028 0.4400 0.54 0.46 0.04
Brain 21 0.02 0.77 0.051 0.0565 0.0028 0.8097 0.99 0.86 0.02
Gut 21 0.0171 0.718 0.0487 0.0163 0.0028 0.7296 0.89 0.77 0.01
Heart 21 0.0047 0.758 0.0115 0.0166 0.0028 0.7697 0.94 0.81 0.00
Kidney 21 0.0044 0.783 0.0207 0.01622 0.0028 0.7944 0.97 0.84 0.00
Liver 21 0.026 0.751 0.0348 0.0252 0.0028 0.7688 0.94 0.81 0.02
Lung 21 0.0076 0.811 0.003 0.009 0.0028 0.8173 1.00 0.86 0.01
Muscle 21 0.4 0.76 0.0238 0.0072 0.0028 0.7651 0.93 0.81 0.32
Skin 21 0.0371 0.718 0.0284 0.0111 0.0028 0.7259 0.88 0.77 0.03
Spleen 21 0.0026 0.788 0.0201 0.0198 0.0028 0.8019 0.98 0.85 0.00
Plasma 21 0.0424 0.945 0.0035 0.00225 0.0028 0.9466 0.525i
Plasma 22 0.0424 0.945 0.0035 0.00225 6E-05 0.9466m
Whole blood 21 0.0771 0.82 0.0032 0.002 0.0028 0.8214l
Whole blood 22 0.0771 0.82 0.0032 0.002 6E-05 0.8214k
Erythrocytes 21 0.0347 NU NU NU
All tissue less plasma 0.956
Parameters for Carbamate Models

a
Values for monoprotic acids. Used in Eq. 15 to calculate D*vo:w.
b
Equation number used to calculate partition coeofficients:
fi Pt:b and Pt:pp.
c
Values used in Eqs. 21 and 22 as presented in Poulin and Thiel (2002a). Values for Eq 6 in Poulin and Thiel (2002a) are: E : P (erythrocyte:plasma in vivo) = 1, B : P
(blood : plasma in vitro) = 1, and Ht (hematocrit in blood) = 0.45.
d
Ko:w as calculated in Eq. 14 .
e
Numerator value from Eq. 21 or 22 depending on tissue.
f
Denominator value used in Eq. 21 or 22 for Pt:b in the rat.
g
Denominator value used in Eq. 21 or 22 for Pt:pp in the rat.
h
Denominator value used in Eq. 21 or 22 for Vt*Pt:p in the rat.
i
Volume distribution at steady state (Vdss; L kg−1).
j
NU, not used in Poulin and Thiel (2002a).
k
Denominator value used in Eq. 21 or 22 for Pt:b in the human.
l
Denominator value used in Eq. 21 or 22 for Pt:pp in the human.
m
81

Denominator value used in Eq. 21 or 22 for Vt*Pt:p in the human.

82 J.B. Knaak et al.

Fig. 6. Liver : blood and fat : blood partition coefficients

fi of aldicarb and metabolites
as a function of ACD log DpH7.4. Eq. 21 involving ACD log DpH7.4 was used to calcu-
late partition coefficients
fi for liver. Eq. 22 involving the conversion of ACD
log DpH7.4 to log D*o:w using Eq. 13 was used to calculate partition coefficients
fi for fat.
The Henderson–Hasselbach equations were not used. Aldicarb and its metabolites
are identifified in Table 14 (Appendix A).

D. Glucuronidation and Transcellular Transport

In recent years investigators have revealed that metabolism can be altered
by changes occuring in drug transport. According to Benet et al. (2003),
the incorporation of efflux
fl and uptake processes lead to better predictions
of drug clearances from in vitro systems. Of interest is the interaction
between P-glycoprotein and CYP3A4 (effl flux-metabolism alliance); OATPs
(organic anion-transporting peptides) and CYP3A4 (uptake transporter-
metabolism); and MRP2 (multidrug-resistant protein) and UGTs
(UDP-glucuronosyltransferases).
Hepatic transporters are categorized into two groups: efflux
fl transporters
that are localized on the canalicular (apical) membrane, and the uptake
transporters that are found on the sinusoidal (basolateral) membrane (Lam
and Benet 2004) of hepatocytes. According to Chang and Benet (2005), the
Vmax and Km constants for glucuronidation by human liver microsomes are
20.2 nmol min−1 mg−1 and 216 μM, respectively, with the exit of 1-naphthyl
glucuronic acid being a slower or faster (not determined) process. Endog-
enous transporters in conjunction with partition coefficients
fi are believed to
 Parameters for Carbamate Models 83

Fig. 7. Liver : blood and fat : blood partition coeffi

ficients of carbaryl and metabolites
as a function of ACD log DpH7.4. Equation 21 involving log DpH7.4 was used to calcu-
late partition coefficients
fi for liver. Eq. 22 involving the conversion of ACD log DpH7.4
to log D*vo:w using Eq. 13 was used to calculate partition coefficients
fi for fat. The
Henderson–Hasselbach equations were not used. Carbaryl and its metabolites are
identifi
fied in Table 15 (Appendix A).

be active in determining the distribution of formed metabolites. The impor-

tance of the activities of transporters and metabolizing enzymes in the
absorption, distribution, metabolism, and elimination (ADME) of carba-
mate pesticides has yet to be established. Inhibitors of these systems may
be used to study their effect on ADME (Lau et al. 2003).

E. Metabolic Enzymes, P450s, and CaEs

Cytochrome P450s
CYP450 was first shown to be a hemoprotein by Omura and Sato (1964).
It has a noncovalently bound iron protoporphyrin IX prosthetic group,
similar to the heme in b-type cytochromes, hemoglobin, and myoglobin.
The heme prosthetic group may be removed from the protein by acidic
acetone, alkali, or pyridine. The binding of carbon monoxide provides a
means for their analysis and quantifi fication. The enzymes of the CYP-
dependent monooxygenase system are embedded in the endoplasmic retic-
ulum of liver hepatocytes and the cells of other organs. The endoplasmic
84 J.B. Knaak et al.

reticulum is harvested from a cell homogenate by centrifugation at 16,000 g

to remove large particles and then at 105,000 g to pellet the microsomes
(endoplasmic reticulum) from the supernatant (Lu and West 1980). The
P450 enzymes are distributed between the two surfaces of the microsomal
bilayer with the NADPH-CYP reductase and P450 facing the cytoplasm of
the cell. Therefore, the monooxygenase system is composed of three com-
ponents: reductase protein, a lipid fraction, and CYP protein. The purifica-
fi
tion of rat and human liver P450s is described by Guengerich and Martin
(1998). An estimation of protein concentrations provides a means for evalu-
ating the progress of purification.
fi The content of CYP heme should be
about 18 nmol mg−1 protein.

Multiple Forms of CYPs

Multiple forms of CYPs exist in liver microsomes. These forms play a role
in the oxidation (i.e., hydroxylation of aromatic, aliphatic, and alkyl group-
ings) of carbamate pesticides. The major isoforms in human liver include
CYP 1A2, 2A6, 2B6, 2C8, 2C9, 2C19, 2D6, 2E1, 3A4, 3A5, and 3A7 (fetal
livers). There are large individual variations in the microsomal content of
these forms in human liver. A calculator for determining the content of
these P450 isozymes in human liver (ages 1–18 yr) was recently presented
by Foxenberg et al. (2006, 2007). The recombinant human CYPs are avail-
able from a Baculovirus-insect cell expression system along with the coex-
pression of NADPH-CYP reductase (Supersomes; Gentest, Woburn, MA).
Yields range from 50 to 250 pmol P450 mg−1 microsomal protein as deter-
mined by CO difference spectral analysis (Hood et al. 1998).

In Vitro Metabolism of Aldicarb, Carbaryl, and Carbofuran by Human

and Rat Liver Microsomes and Individual CYPs
Aldicarb was converted by rat liver microsomes to aldicarb sulfoxide
(Pelekis and Krishnan 1997) with Vmax and Km values of 5.41 nmol min−1 mg−1
microsomal protein and 184 μM, respectively. Negligible quantities of aldi-
carb sulfone were found after incubation. The results imply that under in
vivo conditions aldicarb sulphoxidation is not likely to be saturated at lethal
dose levels. CYP and the fl
flavin monooxygenase systems (FMO) are involved
in this process (Perkins et al. 1999). Selective in vitro inhibition of flavin-
containing and CYP monooxygenases confi firmed that the former enzymes
mainly catalyze sulfoxide production whereas the latter catalyze sulfone
formation (Montesissa et al. 1994).
Three major metabolites of carbaryl (i.e., 5-hydroxy-, 4-hydroxy-, and
methylolcarbaryl) were formed by human liver microsomes or individual
CYP isoforms (Tang et al. 2002). CYP1A1 and 1A2 formed 5-hydroxylcar-
baryl while CYP3A4 and 1A1 were the most active in forming 4-hydroxy-
carbaryl. Activity assays were performed by the incubation of carbaryl (up
to 500 μM) with microsomes (1 mg protein) or CYPs (18–50 pmol P450) in
 Parameters for Carbamate Models 85

the presence of an NADPH-generating system, 100 mM Tris (pH 7.4) buffer

containing 5 mM MgCl2 for 15 min. In the case of the CYPs the buffers used
were recommended by Gentest. Analysis was performed using HPLC and
analytical standards.
Kinetic assays used carbaryl concentrations ranging from 10 to 1,000 μM
under the conditions described above. Vmax and Km values for HLMs and
individual CYPs are given in Appendix B, Tables 34 and 35 for carbaryl.
1-Naphthol was not a major product of CYP-catalyzed reactions in the
presence or absence of NADPH. Conditions did not appear to be favorable
for the hydrolysis of carbaryl by CaEs. Carbaryl is believed to be both an
inhibitor and a substrate for these enzymes. In Table 34, the in vitro car-
baryl HLM Vmax values (μmol min−1 mg−1 protein) may be expressed in vivo
values (μmol hr−1 kg−1 bw). This calculation is accomplished by multiplying
the in vitro values by 60 min hr−1; 30 mg microsomal protein/g of liver; and
by 27 g liver/kg bw to give the in vivo value. In Table 35, the in vitro CYP
values for carbaryl are expressed in nmol min−1 nmol−1 P450. These values
may be converted to in vivo values (nmol hr −1 kg−1 bw) by multiplying the
values by CYP content in nmol mg−1 microsomal protein, 60 min hr −1; 30 mg
microsomal protein/g liver; and by 27 g liver/kg of bw to give the in vivo
value.
The effects of carbaryl were investigated in conjunction with microsomal
hepatic lipid peroxidation, and NADPH-dependent reductase activities.
Carbaryl did not affect lipid peroxidation under in vivo conditions. More-
over, following administration of the compound, the activities of NADPH-
cytochrome reductase as well as NADPH-neotetrazolium reductase was
not inflfluenced by carbaryl (Beraud et al. 1989).
In a paper by Cho et al. (2006), CYP3A4 converted 1-naphthol to 1,4-
dihydroxydihydronaphthalene (4-OH naphthol) with Vmax, Km values of
(1.58 × 10−3 μmol min−1 mg−1 protein), 77 μmol hr−1 kg−1 bw, and 80 μM, respec-
tively. A number of other metabolites were found but not identified. fi These
metabolites may be 2-OH naphthol, 5-OH napththol, 3,4-dihydroxydihy-
drol naphthol, and 5,6-dihydroxydihydrol naphthol in Appendix B under
carbaryl. On the basis of protein content in the liver CYP3A4, 1A2, and
2C19 were considered the most important isoforms for 1-naphthol
metabolism.
Usmani et al. (2004a) studied the in vitro metabolism of carbofuran
by human liver microsomes (HLMs), mouse liver microsomes (MLMs),
and rat liver microsomes (RLMs), and CYPs to 3-OH carbofuran (and
two minor metabolites). Km values were 1.974, 0.207, and 0.551 mM for
HLMs, RLMs, and MLMs, respectively. The Vmax values were 3.30 and
5.50 nmol min−1 mg−1 protein for HLMs and RLMs. The intrinsic clearance
rates for RLMs and MLMs were 14 fold greater than HLMs. CYP3A4 is
the major isoform responsible for the oxidation of carbofuran in humans,
with CYP1A2 and 2C19 possessing some but less activity. The activity of
HLMs from 17 single donors varied by more than 5 fold, indicating that the
86 J.B. Knaak et al.

specific
fi content of at least CYP3A4 was not constant in these microsomes.
The HLM and individual CYP Vmax and Km values for carbofuran are sum-
marized in Tables 34 and 35, respectively.
Pretreatment of rats with tetraisopropylpyrophosphoramide (iso-
OMPA) (CAS no. 513-00-8) 1 hr before carbofuran potentiated the toxicity
of this N
N-methylcarbamate insecticide threefold. CaE activity in a variety
of organs including brain, muscle, liver, and plasma was signifi ficantly
reduced, while acetylcholinesterase (AChE) activity was unchanged. Sig-
nificant
fi inhibition of AChE was observed after the combination of iso-
OMPA and carbofuran (Gupta and Dettbarn 1993).
Buronfosse et al. (1995) showed the sulfoxidation of methiocarb is cata-
lyzed by FMO and CYP with 50% being carried out by each enzyme system.
Stereoselective sulfoxidation occurs only for FMO with an enantiomeric
excess of 88% in favor of the (A)-enantiomer. Lung and kidney microsomes
have high flavin-containing monooxygenase levels. FMO is important rela-
tive to CYP in these tissues. Thioether-containing organophosphates are
effective substrates for the fl
flavin-containing monooxygenase in mouse liver
microsomes, with Km values between 3.5 and 36 nM. According to Tynes
and Hodgson (1985), thioether-containing carbamates are less effective
substrates in other animals having Km values near 280 nM (Tynes and
Hodgson 1985). The sulfoxidation of thioethers by HLMs was found to be
predominantly metabolized by P450 isozymes (85%–90%) compared with
FMO isoforms. The highest rates were obtained with CYP1A1, 1A2, 3A4,
2B6, 2C9*1, 2C19, 2D6*1, and FMO1 (Usmani et al. 2004b).
Very little if any work has been done on the in vitro metabolism of
oxamyl in humans. In vitro rat liver studies were carried out by Harvey and
Han (1978). NADPH fortified fi microsomal plus soluble rat liver fractions
failed to yield N-demethylated oxamyl oxime or oxamyl acid. The data
supports the N-demethylation of oxamyl as the pathway to des N-methyl
N
oxime or des NN-methyl oxamyl acid.

Hydrolysis of Carbamates by CaEs

Mammalian CaEs are located in the endoplasmic reticulum, cytosol, and
mitochondria of cells. Hosokawa et al. (1990) compared the liver micro-
somal CaE activity in nine animal species and humans. A review of mam-
malian carboxylesterases by Satoh and Hosokawa (1998) lists (in table 1)
the catalytic activity of highly purified
fi CaE isozymes from mammalian liver
microsomes. Microsomal CaEs hydrolyze p-nitrophenylacetate, malathion,
butanilicaine (CAS no. 3785-21-5), and isocarboxazid (CAS no. 59-64-0)
with RLMs having the highest specifi fic activity and humans the lowest.
Activity was reported in μmol min−1 mg−1 of microsomal protein.
CaEs are believed to be involved in the hydrolysis of the carbamic ester
bond of pesticidal carbamates (Gupta and Dettbarn 1993). 1-Naphthol ([S],
125 μM carbaryl) was generated at the rate of 0.034 nmol min−1 mg−1 protein
with pooled HLM in the presence and absence of an NADPH-regenerating
 Parameters for Carbamate Models 87

system (Tang et al. 2002). NADPH was used to determined whether an

oxidative enzyme was involved in the hydrolysis. This rate is equivalent
to 1.65 μmol hr−1 kg−1 body weight. 1-Naphthol was also formed by cyto-
solic enzymes from carbaryl (125 μM) at a rate of 0.008 nmol min−1 mg−1
protein. These rates are low when compared to rates (specific fi activity,
9.1 μmol min−1 mg−1 protein; Km, 12.1 μM) obtained using p-nitrophenyl
acetate as substrate (Stok et al. 2004).
Ross et al. (2006) determined the activity of recombinant CE (i.e., CaE)
and liver microsomes by measuring the production of p-nitrophenol liber-
ated from p-nitrophenyl valerate, acetate, or butyrate ([S], 500 μM) at
400 nm on a spectrophotometer. An extinction coefficient
fi of 13 cm−1 mM−1
was used to convert the slopes of each activity curve to specific fi activities.
Specifific activities for CaEs in rat and pooled HLMs were 9.33 and
2.21 μmol min−1 mg−1 protein, respectively, for p-nitrophenyl acetate. The
kinetic parameters (Vmax, nmol min−1 mg−1; Km, μM) for the hydrolysis of
trans-permethrin by RLMs and pooled HLMs were, respectively, 1.34,
19.16 and 1.12, 20.66. CaE activity was 39 times greater toward trans-
permethrin than toward carbaryl. The hydrolysis of carbaryl may have been
reduced by the inhibitory effects of carbaryl on CaE (Stok et al. 2004).
Rabbit serum albumin catalyzes the hydrolysis of carbaryl (kcat, 7.1 ×
10−5 sec−1 or 0.256 hr−1; Km, 240 μM) and p-nitrophenyl butyrate, with butyr-
ate ester being a competitive inhibitor of the hydrolysis of carbaryl (Sogorb
et al. 2002). The inhibition of carbaryl hydrolysis by sulfhydryl-blocking
agents suggests that a cysteine residue plays an important role in the active
center. Human, chicken, and bovine serum albumins are also able to hydro-
lyze carbaryl.

F. Models for In Vivo Metabolism of Carbamates

The metabolic pathways for the 10 carbamates of interest are given in
Tables 24–33 (see Appendix B). The tables were developed from pathway
data in published literature and from physiolog ical models developed from
the pathways. A physiolog ical model for aldicarb is presented in Appendix
C. Similar physiolog ical models were developed for each of the remaining
9 carbamates but are not included in Appendix C. ACSL programs were
written for each physiolog ical model. The models may be used to develop
graphic models using ACSL’s Graphic Modeler. ERDEM is an example of
a graphic model. The metabolic pathway data in Appendix B may be used
in command files for programming ERDEM. The following paragraphs
briefl
fly describe the metabolic information found in the literature for the 10
carbamates of interest.

Aldicarb
The metabolic pathway is described in Table 24 (Appendix B). This carba-
mate pesticide is readily biotransformed by animals and humans (Andrawes
et al. 1967; Baron 1994; Knaak et al. 1966; Marshall and Dorough 1977,
88 J.B. Knaak et al.

1979; Tobia et al. 2004). Urinary metabolic products of aldicarb include the
more toxic aldicarb sulfoxide (∼35%), the less toxic sulfone (∼1.0%), oxime
sulfoxide (∼35.5%), oxime sulfone (∼2.0%), and neutrals or polar metabo-
lites (∼30.5%) as the major products (Knaak et al. 1966). The neutrals are
most likely nitrile sulfoxide and sulfone and alcohol sulfoxide and sulfone.
The structures of the metabolites are listed in Table 14 (see Appendix A).
Andrawes et al. (1967) reported that 80% of an oral dose to rats was elimi-
nated in urine and 4% in feces. Approximately 50% of the urinary metabo-
lites were water soluble and could not be extracted into organic solvents.
The hydrolysis products, aldicarb oxime, oxime sulfoxide and oxime sulfone,
have no significant
fi toxicity to animals or insects. The nitriles come from the
degradation of aldicarb, aldicarb sulfoxide and aldicarb sulfone and are the
primary stable products of aldicarb degradation (Cobb et al. 2001). Eigh-
teen analytical standards were prepared by Durden et al. (1970) for studies
involving the metabolism of aldicarb in plants and animals.

Carbaryl
The metabolic pathway is described in Table 25 (Appendix B). The princi-
pal metabolic pathways of carbaryl are ring hydroxylation and hydrolysis
(Dorough and Casida 1964; Knaak et al. 1965; Knaak and Sullivan 1967;
Knaak et al. 1968). As a result, numerous metabolites (aglycones) are
formed and subjected to conjugation with the formation and urinary
excretion of water-soluble sulfates and glucuronides (Cardona and
Dorough 1973; Knaak et al. 1967). Hydrolysis results in the formation of
1-naphthol, carbon dioxide, and methyl-amine. Hydroxylation produces
4-hydroxycarbaryl, 5-hydroxycarbaryl, N N-hydroxy-methylcarbaryl, 5,6-
dihydro-5,6-dihydroxycarbaryl, and 1,4-naphthalendiol (Leeling and
Casida 1966; Sullivan et al. 1972). The principal metabolites in humans
are the glucuronides of 1-naphthol and 4-hydroxycarbaryl and the sulfate
of 1-naphthol (Knaak et al. 1968). Chen and Dorough (1979) found no
evidence for the conjugation of naphthyl-14C-carbaryl with glutathione or
mercapturic acid in their studies. The hydrolytic pathway appears to be a
major pathway in humans with ring hydroxylation of either 1-naphthol or
carbaryl as secondary pathways (Knaak et al. 1968). Under normal expo-
sure conditions, the accumulation of carbaryl in animals is unlikely.
Enterohepatic cycling of carbaryl metabolites in rats has been reported
by Marshall and Dorough (1979) and Struble et al. (1983). According to
Marshall and Dorough (1979), 37.5% of an oral dose (0.01 mg kg−1 bw)
was excreted in bile after 3 hr, principally as glucuronides. The glucuronides
are absorbed, metabolized, and recycled, resulting in 1.4% of the dose
ultimately being eliminated in feces. Metabolites identified fi in bile were
5,6-dihydro-5,6-dihydroxycarbaryl glucuronide (12%–18% of biliary 14C),
and the conjugated isomer of hydroxymethylcarbaryl (2% of biliary 14C).
The percentage of biliary 14C decreased with higher dosages, with doses
 Parameters for Carbamate Models 89

of 1.5–30.0 mg kg−1 bw resulting in three times more biliary 14C than a

300 mg kg−1 bw dose (Struble et al. 1983).
Carbaryl, 1-naphthol, and their hydroxylated metabolites were sepa-
rated on thin-layer glass plates or silica gel columns and identified fi by
cochromatography with known standards. The conjugates, glucuronides
and sulfates, were separated by weak ion-exchange chromatography (i.e.,
DEAE cellulose) and identified fi by cochromatography with known stan-
dards (Knaak et al. 1965). Separation of closely related conjugates (i.e.,
glucuronides of 4-OH and 5-OH carbaryl) was difficultfi to achieve. In many
cases it was necessary to hydrolyze the conjugates using glucuronidase or
sulfatases and to identify the aglycones using silica gel column or thin layer
chromatography. Knaak et al. (1967) used GC to separate several glucuro-
nides on a SE-30 column as the acetylated or silylated derivatives of their
methyl esters. In practice, methylation was carried out after acetylation,
whereas in the case of the silylated derivatives, methylation was performed
first followed by silylation.
fi

Carbofuran
The metabolic pathway for carbofuran is described in Table 26 (Appendix
B). The information on the pathway was extracted from Dorough (1968),
Ferguson et al. (1984), Fukuto (1972), Ivie and Dorough (1968), Knaak
et al. (1970), Knaak (1971), Marshall and Dorough (1979), Metcalf et al.
(1968), Roberts and Hutson (1999), and Usmani et al. (2004a).
The fate of ring-C14 and carbonyl-C14 carbofuran was studied by Knaak
et al. (1970) in the dairy cow with 83% and 12% of the dose, respectively,
being eliminated in urine. Less than 4% of the administered ring-labeled
carbofuran was eliminated in feces. The major metabolites were the sulfates
of carbofuran phenol (45.4%) and 3-keto-7-phenol (10.4%) and the gluc-
uronides of carbofuran phenol (14.4%), 3,7-diol (15.9%), and 3-hydroxy
carbofuran (4.2%). 3-Hydroxycarbofuran-carbonyl-C14 is hydrolyzed in the
cow to almost the same extent as carbonyl-C14-carbofuran.
Ivie and Dorough (1968) separated the urinary metabolites of ring-C14
carbofuran from the cow into organo extractables and water solubles, with
90%–97% of the label being present as water solubles. Carbofuran phenol
(34.36%) was the major hydrolysis product recovered in the water phase
of a 12-hr urine after acid hydrolysis along with 3-keto-7-phenol and the
3,7-diol. Small quantities of 3-OH carbofuran (9.14%) and 3-OH-N- N
hydroxymethyl carbofuran (2.16%) were also reported. Acid hydrolysis
failed to completely release the aglycones of carbofuran from their conju-
gates, leaving up to 57% in the water phase. The differences reported
between the two cow studies appear to result from (1) the administration
of carbofuran orally by bolling gun, (2) the administration via a rumen
fistula, and (3) the analytical procedures used to determine the urinary
fi
metabolites.
90 J.B. Knaak et al.

In rats, oxidative and hydrolytic metabolites accounted for about 70%

and 21%, respectively, of the administered radioactivity (Roberts and
Hutson 1999). Oxidative metabolism was the predominant pathway in mice
(Dorough 1968). Ferguson et al. (1984) presented tissue-time course data
for the disappearance of carbofuran and the appearance of 3-OH carbofu-
ran in the rat after the oral and intravenous administration of carbonyl-C14-
labeled carbofuran. Organoextractable metabolites were analyzed using
thin-layer chromatography (TLC), while water-soluble conjugates were
first hydrolyzed and then analyzed by TLC. No information was obtained
fi
by Ferguson et al. (1984) regarding the fate of the hydrolyzed products
(i.e., carbofuran phenol, 3,7-diol, and 3-keto-7-phenol).
According to Marshall and Dorough (1979), biliary elimination of gluc-
uronide metabolites amounted to 2.6% of dose 3 hr after dosing and 28.5%
after 48 hr. The glucuronides, principally 3-OH carbofuran glucuronide,
were absorbed from the gastrointestinal tract, metabolized, and eliminated
in urine or absorbed back into bile to be recycled. The overall result was
the elimination of 4% of the dose in feces and the remainder in urine.

Formetanate
The metabolic pathway for formetanate is described in Table 27 (Appendix
B). Information on the pathway was taken from Sen-Gupta and Knowles
(1970), Fukuto (1972), and Roberts and Hutson (1999).
In rats treated with formetanate, 60%–80% of the urinary excretory
products were water-soluble conjugates. The major pathway was the forma-
tion of the formamino derivative, 3-formaminophenyl N-methylcarbamate,
N
followed by the enzymatic removal of the NN-methylcarbamoyl group to give
formaminophenol. Deformylation of formaminophenol (Ahmad and
Knowles 1971) resulted in 3-aminophenol whereas subsequent N-acetyla-
tion produced 3-acetamidophenol.
Major aglycones of sulfuric and glucuronic acids were 3-acetamidophe-
nol, 3-formaminophenol, and 3-aminophenol. Water-soluble carbamates
include 3-formaminophenyl-N-methylcarbamate
N and demethylformetanate.
Sen-Gupta and Knowles (1970) found 3-aminophenyl N-methylcarbamate
N
and formetanate phenol in plants but did not fi find them in rat urine. An
oral dose of 14C-formetanate was rapidly absorbed and excreted by rat, 85%
in urine and 8% in feces. After 72 hr, 2% of the 14C was still retained.

Methiocarb
The metabolic pathway of methiocarb is described in Table 28 (Appendix
B). The information was taken from Kuhr and Dorough (1976), Menzie
(1974), Oonnithan and Casida (1968), Roberts and Hutson (1999), Strother
(1972), and Van Hoof and Heyndrickx (1975).
When rat and human liver enzymes were incubated with [14C-carbonyl]-,
[ C-4-methyl]-, or [14C-N-methyl]
14
N methiocarb, about a dozen metabolites
 Parameters for Carbamate Models 91

were observed. The major metabolite was methiocarb sulfoxide, with the
human liver producing slightly less than the rat (13% vs. 16%), according
to Strother (1972). Hydrolysis yielded the phenolic moiety of methiocarb,
oxidation of methiocarb sulfoxide yielded the sulfone, while hydroxylation
produced N N-hydroxymethyl methiocarb. Products were eliminated in free
and conjugated forms. Several metabolic products from human liver studies
were not observed in rat studies (Strother 1972). Rats given a single oral
dose of 5.0 mg methiocarb eliminated up to 2.3% of the dose in the urine
as unchanged methiocarb and 3.3% as its phenolic metabolites, mostly
within 48 hr of administration (Hayes and Laws 1991).

Methomyl
The metabolic pathway of methomyl is described in Table 29 (Appendix
B). Information on the metabolic pathway was taken from Harvey et al.
(1973), Huhtanen and Dorough (1976), and Reiser et al. (1997). In the
study by Harvey et al. (1973), radiolabeled methomyl (S-methyl [1-14C]
N-[(methylcarbamoyl)-oxy] thioacetimidate) administered orally to rats is
N
metabolized and rapidly eliminated within 24 hr in the ratio of one part
[14C]carbon dioxide, two parts [1-14C]acetonitrile, and one part urinary
metabolites. The identities of the urinary metabolites were not established
by Harvey et al. (1973); however the absence of methomyl, S-methyl
N-hydroxythioacetimidate, the S-oxide of methomyl, and the S,S-dioxide
N
of methomyl and conjugates was demonstrated. Methomyl can exist in two
geometric configurations
fi (Huhtanen and Dorough 1976), Z and E isomers.
In rats, carbonyl- or oximino-labeled syn-methomyl (Z isomer) was metab-
olized to carbon dioxide and acetonitrile at a 2 : 1 ratio. In contrast, the anti
isomer (E isomer) was metabolized predominantly to acetonitrile. It was
concluded on the basis of this and other evidence that a Beckman rear-
rangement of the syn- and anti oximes occurs before formation of carbon
dioxide and acetonitrile.
The methomyl PBPK/PD model developed in this review, but not pre-
sented, contains a lung compartment for removing 14C-carbon dioxide and
acetonitrile formed during the metabolism of methomyl in the liver. The Z
isomer of methomyl is partially converted to the E isomer during metabo-
lism. The Z isomer is metabolized to carbon dioxide, while the E isomer
goes to acetonitrile. AChE inhibition is believed to be largely caused by
the Z isomer, although the E isomer may also produce inhibition.
The acetonitrile formed by hydrolysis of methomyl is conjugated with
glutathione. The glutathione conjugate is hydrolyzed to form the glutam-
ylcysteine, cysteinylglycine, and the cysteine conjugates of acetonitrile. The
cysteine conjugate goes on to form an N N-sulfate–cysteine conjugate (Reiser
et al. 1997).
The main metabolites of methomyl are acetonitrile per se (5%–34% of
dose), respiratory carbon dioxide (19%–34% of dose), and unknown urinary
92 J.B. Knaak et al.

metabolites in a ratio of 2 : 1 : 1. One of the urinary metabolites (1%–4% of

dose) was identified
fi as the N N-sulfate–cysteine conjugate. A large number
of the urinary metabolites are unknown products.

Oxamyl
The metabolic pathway for oxamyl is described in Table 30 (Appendix B).
This information was extracted from Chang and Knowles (1979) and Harvey
and Han (1978). Oxamyl is degraded by three pathways, hydrolysis to the
oxamyl oxime (oximino metabolite), des N N-methyl oxamyl, or enzymatic
conversion via oxamyl nitrile (DMFC) to oxamyl acid (N,
N N-dimethyloxamic
N
acid). The oxime and acid are both conjugated with glucuronic acid along
with des NN-oxamyl oxime and des N-oxamyl acid (N-methyloxamic acid)
from des N-methyl oxamyl before elimination in urine and feces (>70%)
(Harvey and Han 1978). No oxamyl or other organo-soluble metabolite was
detected in urine, feces, or tissues. 14C from oxamyl oxime is extensively
converted to 14CO2 or incorporated into amino acids accounting for most
(>50%) of the radioactivity retained in the tissues. Similar products were
found in the urine and feces of the mouse from intraperitoneally (IP)-
administered oxamyl-14C.
In the intact rat administered 14C-oxamyl (2.5–4.6 mg kg−1 bw), most of
the dose (68%–72%) was recovered in urine with lesser amounts in feces
after 72 hr. Conjugates of the oximino compound, the acid, and their mono-
methyl derivatives constituted >70% of the metabolites excreted in the
urine and feces. Less than 0.3% of the oxamyl was exhaled as carbon
dioxide (Harvey and Han 1978). Mice treated IP with 14C-oxamyl elimi-
nated 96.4% of the dose by 96 hr, 88.7% in urine and 7.7% in feces (Chang
and Knowles 1979).
Extensive degradation/metabolism of (1-14C) oxamyl was observed in
the goat. Radioactive thiocyanate was the major metabolite identified fi
in milk as well as in the methanol/water extracts for all tissue samples.
Oxamyl-derived residues in the urine derived from oxamyl nitrile have
been identified
fi as thiocyanate, des N N-oxamyl acid, oxamide, and des N-N
methyloxamide (Li et al. 1997). Thiocyanate is generated from cyanide
through the reaction with thiosulfate catalyzed by rhodanese (Solomonson
1981). The metabolic pathway in the ruminant (goat) appears to be signifi-fi
cantly different from the pathways determined in monogastric animals such
as the rat and mouse.

Pirimicarb
The metabolic pathway of this dimethylcarbamate is described in Table 31
(Appendix B). Information on the pathway was taken from Baron (1991).
Pirimicarb’s major urinary metabolites in the rat, dog, and cow are similar,
resulting from oxidative and hydrolytic mechanisms, and consisting pri-
marily of hydroxypyrimidines with modifi fications of the alkyl constituents
 Parameters for Carbamate Models 93

of the heterocyclic moiety. Of the administered dose, 2-dimethylamino-5,

6-dimethyl-4-hydroxypyrimidine (DDHP) accounted for 10%–16.3%, 2-
methylamino-5,6-dimethyl-4-hydroxypyrimidine (MDHP) for 20.5%–41%,
2-amino-5,6-dimethyl-4-hydroxypyrimidine (ADHP) for 12.9%–21%, and
2-dimethylamino-6-hydroxymethyl-5-methyl-4-hydroxypyrimidine
(DHHP) for 1.8%–5.7%. The major metabolites were eliminated unconju-
gated. The glucuronide of pirimicarb phenol and hydroxymethyl pirimicarb
are included in Table 31 (Appendix B) as possible metabolites. The gluc-
uronide is more water soluble than the phenol at pH 7.4.
DDHP, MDHP, and ADHP were detected in urine samples of seven
workers who had applied pirimicarb (Hardt et al. 1999; Hardt and Angerer
1999). Concentrations of MDHP and ADHP were much higher than that
of DDHP, indicating a considerable demethylation capacity in humans. No
metabolites were found in urine specimens of controls. The pyrimidines
investigated represent sensitive and specifi
fic parameters for biological moni-
toring of exposure to pirimicarb (Hardt et al. 1999).
In an earlier metabolism study involving [1,3-15N-2-2- N 13
C]-labeled
14
pirimicarb mixed with 2-C -labeled pirimicarb, 10 metabolites were
found in the excreta of rats (Hendley and Lam 1981). The metabolites
were DHHP, MDHP, ADHP, 2-[(methoxymethyl) methylamino)-4-
hydroxy-5,6-dimethylpyrimidine (CAS no. 78195-32-1), 2-[(methoxymethyl)
amino]-4-hydroxy-5,6-dimethylpyrimidine (CAS no. 78195-33-2), and
2-[(hydroxymethyl)methylamino]-4-hydroxy-5,6-dimethylpyrimidine
(CAS no. 78195-31-0). The last three metabolites were formed by hydrox-
ylation of a methylamino group, methylation of the hydroxy group, and
demethylation of an N N-methyl group, but were not found in the human
studies (Hardt et al. 1999). Their log DpH 7.4 values, 0.15, −0.06, and −0.49,
respectively, suggest that these metabolites are more water soluble than
pirimicarb phenol, and if present would appear in urine unconjugated.

Propoxur
The metabolic pathway for propoxur is described in Table 32 (Appendix
B). Information on the pathway was obtained from the in vitro work of
Oonnithan and Casida (1968) and the review of Knaak (1971). RLMs, in
the presence of NADPH, converted carbonyl-14C, N-methyl-
N 14
C, and ring-
14
C-propoxur to seven metabolites possessing the intact carbamate and
isopropoxy group, 2-isopropoxy-4-hydroxyphenyl methylcarbamate,
2-isopropoxy-5-hydroxyphenyl methylcarbamate, 2-isopropoxyphenyl
N-hydroxymethylcarbamate, and unidentifi
N fied products. The one metabo-
lite with the phenyl O−C (O)−N−C linkage was 2-hydroxyphenyl meth-
ylcarbamate, whereas the three phenols with the isopropoxy group (C−(C)
C−O) were 2-isopropoxyphenol, 2-isopropoxy-4-OH phenol, and 2-isopro-
poxy-5-OH phenol. In the presence of UDP-glucuronic acid, these products
would be conjugated with glucuronic acid. Sulfates appeared in the urine
94 J.B. Knaak et al.

of rats after the 2-wk daily oral administration of 30–50 mg kg−1 bw of pro-
poxur (Foss and Krechniak 1980; Krechniak and Foss 1982, 1983). At least
one of the unidentifi fied metabolites and 2-isopropoxy-5-hydroxyphenyl
methylcarbamate are more potent ChE inhibitors than propoxur (Oonni-
than and Casida 1968).
Propoxur is biotransformed in vivo by depropylation to 2-hydroxyphe-
nol-N-methyl-carbamate,
N and by hydrolysis to the phenol. Ring hydroxyl-
ation and isopropoxy hydroxylation followed by conjugation account for
the glucuronides detected in urine (ACGIH 1991). The in vivo fate of
individual 7.5 μmol kg−1 bw dosages (IP) of carbonyl-14C and N-methyl-14C
propoxur in the rat were studied by Krishna and Casida (1966). 14C-CO2
was recovered in exhaled air from the carbonyl-(31.2%) and methyl-
(21.7%) labeled propoxur after 48 hr, with the remaining percentage being
recovered in urine (>60%), feces (∼2%), and body tissues (∼2%–5%).
According to Hayes and Laws (1991), the relationship of urinary excre-
tion of 2-isopropoxyphenol to propoxur concentration was nearly linear
following inhalation exposure. This metabolite was found in the urine 3 d
after termination of exposure to 78 mg m−3, 2 d after exposure to 9 mg m−3,
and 24 hr after exposure to 0.4 mg m−3.

Thiodicarb
The metabolic pathway is described in Table 33 (Appendix B). Information
on the pathway was obtained from WHO/FAO (2000), and the metabolic
work on methomyl in Table 29 (Appendix B) (Harvey et al. 1973; Huhtanen
and Dorough 1976; Reiser et al. 1997).
Thiodicarb (acetamide-14C) is rapidly absorbed from the gastrointestinal
tracts of rats, metabolized to unstable intermediates (i.e., methomyl, metho-
myl oxime, etc.), and finally
fi to acetonitrile, CO2, and low molecular weight
metabolites. The initial step in the process involves hydrolysis to methomyl.
The nature of the isomer, either (E) or (Z), was not determined. In the rat,
an oral dose of 2.0 mg kg−1 bw thiodicarb (acetamide-14C) reached a peak in
plasma after 10 hr and was eliminated in urine (30%) and exhaled air
(40%). The urine contained water-soluble materials while the exhaled air
contained radiolabeled carbon dioxide and acetonitrile. The overall
NOAEL for erythrocytic and splenic effects was 3 mg kg−1 bw/d in a 79-wk
study (WHO/FAO 2000). Brain ChE depression (>60%) occurred at
5 mg kg−1 bw, with pin-point pupils and reduced body temperature.

G. Response: In Vivo antiChE activity

The development of antiChE time course data for carbamates has relied
heavily upon the administration of the carbamate (e.g., oral, dermal, inhala-
tion) followed by the determination of ChE activity in tissues such as the
blood and brain at selected time intervals using spectrophotometric assays
such as the Ellman assay (Ellman et al. 1961). Under normal Ellman assay
 Parameters for Carbamate Models 95

conditions, significant
fi spontaneous reactivation occurs with carbaryl
(Nostrandt et al. 1993). Assay modififications that include (1) preincubation
of concentrated tissue with concentrated chromogen (i.e., DTNB), (2) dilu-
tion to fi
final reaction volume immediately before measurement, and (3)
measurement of ChE over a short period of time (5–10 min) gave results
comparable to a radiometric method. Signifi ficant spontaneous reactivation
may still occur with the modified
fi method if the assay is carried out for
>10 min (Nostrandt et al. 1993).

Aldicarb
The major metabolite of aldicarb, aldicarb sulfoxide, has been shown to
have 76 times greater antiChE activity than the parent compound (DHEW/
NCI 1979). Rats receiving an oral dose (0.33 mg kg−1 bw) of aldicarb
recovered from AChE inhibition 2 hr earlier than rats receiving aldicarb
sulfoxide (Knaak et al. 1966). AChE measurements were made using a
pH Stat. Cambon et al. (1979) studied the effect of aldicarb on the
activity of AChE in tissues from pregnant rats and fetuses. Dosages of
0.001, 0.01, and 0.10 mg kg−1 bw were administered to pregnant rats (oral
LD50 = 1 mg kg−1 bw; Kuhr and Dorough 1976) on day 18 of gestation. Tissue
samples taken from animals at the two highest dosages 1 and 5 hr post
administration showed a significant
fi decrease in AChE activity. The effect
persisted in maternal and fetal blood beyond 24 hr. The lowest dose did not
change AChE activity after 1 hr in maternal tissues except for the liver,
while the AChE activity in fetal blood, brain, and liver were signifificantly
inhibited. AChE determinations were made using the method of Ellman
et al. (1961).

Carbaryl
Short-term studies in animal species confirm
fi that carbaryl can cause toxicity
from ChE inhibition. Wide variations in the dosage required to induce
toxicity in either different species or in one species by different routes of
administration can in part be explained by differences in drug disposition.
Limited long-term exposure studies in rats and dogs have not demonstrated
unexpected adverse effects. However, long-term exposure in pigs results in
a progressive neuromyopathy that is associated with structural damage and
is not acutely reversible with atropine (Branch and Jacqz 1986a,b).
Motor activity and neuromotor function were examined in adult Sprague-
Dawley rats exposed to carbaryl, and behavioral effects were compared
with the time course of ChE inhibition. Rats received an IP injection of 0,
4, 8, 16, or 28 mg kg−1 bw carbaryl in corn oil 20 min before testing. Dosages
of 8, 16, and 28 mg kg−1 bw decreased rat maze activity whereas 16 and
28 mg kg−1 bw reduced open field activity. Maximum effects of carbaryl on
blood and brain ChE and motor activity were seen within 15 min. Maze
96 J.B. Knaak et al.

activity had returned to control levels within 30–60 min, whereas ChE levels
remained depressed for 240 min (Ruppert et al. 1983).

Carbofuran
Cambon et al. (1979) studied the effect of carbofuran on the activity of
AChE in tissues from pregnant rats and fetuses. Dosages of 0.05, 0.25,
and 2.50 mg kg−1 bw were administered to pregnant rats (oral LD50 =
11 mg kg−1 bw; Kuhr and Dorough 1976) on day 18 of gestation. The largest
dose, 2.5 mg kg−1 bw, resulted in signs of AChE inhibition within 5 min.
Eight of 32 rats treated at this level died within 30 min. AChE activity in
maternal brain was 27% of control activity whereas activity in fetal brain
was 80%. AChE activity was signifi ficantly reduced in maternal and fetal
tissues taken from surviving dams 1 hr after treatment at the high dose.
Activity was considered to be normal after 24 hr. AChE activity in tissues
taken from animals at the lower dosages (0.25 and 0.05 mg kg−1 bw) 1 hr after
administration was less than control values in certain tissues. Decreases
were observed in blood from dams and fetuses and maternal liver 1 hr after
the administration of 0.05 mg kg−1 bw. AChE determinations were made
using the method of Ellman et al. (1961).

Formetanate
Phenyl NN-methylcarbamates exert their insecticidal activity by inhibiting
enzyme ChE and do not require activation by metabolism before exhibiting
their enzyme-inhibiting properties (Parke 1968). Moser and MacPhail
(1987) studied the effects of ChE inhibition on operant behavior in rats. A
dose of 0.5 mg kg−1 bw formetanate produced a pronounced suppression of
response rates in trained rats.

Methiocarb
The antiChE insecticide, methiocarb, inhibits serine hydrolases by carba-
mylating a serine residue at the catalytic site. This insecticide is viewed as
a potential inhibitor of serine hydrolase-dependent immune functions,
including interleukin 2 (IL-2) signaling (Casale et al. 1993).

Methomyl
The inhibition of ChE by methomyl is quickly reversed in dogs given
10 mg kg−1 bw methomyl without atropine pretreatment. Signs of intoxica-
tion disappeared in 2 hr, and blood ChE levels returned to normal in less
than 4 hr (ACGIH 2001).

Oxamyl
For subchronic or prechronic exposure in female rats given oxamyl at
dietary levels of 100 ppm or higher for 29 d, AChE activity was decreased
 Parameters for Carbamate Models 97

in the plasma from day 7 on and in the brain at sacrifice. fi There were no
clear effects on erythrocyte AChE levels. In males, AChE activity was
marginally depressed in plasma and erythrocytes. AChE activity was not
affected by oxamyl at 50 ppm (2.5 mg kg−1 bw d−1) (Hayes and Laws 1991).
According to Kennedy (1986), a 4.86 mg kg−1 bw single bolus dose of oxamyl
dropped blood ChE activity in the rat from 4.3 ± 1.0 to 2.6 ± 0.8 μmol sub-
strate hydrolyzed min−1 at 5 min after treatment. Activity fell to 1.8 ± 0.7 μmol
substrate hydrolyzed min−1 at 4 hr after treatment and returned to normal
(5.7 ± 0.8) 24 hr post dosing.

Pirimicarb
Cambon et al. (1979) studied the effect of pirimicarb on the activity of
AChE in tissues from pregnant rats and fetuses. Dosages of 2.0, 20,
and 50–150 mg kg−1 bw were administered to pregnant rats (oral LD50 =
145 mg kg−1 bw; Kuhr and Dorough 1976) on day 18 of gestation. All the
rats dosed with 50 mg kg−1 bw died soon after administration. AChE activity
in the maternal brain was 20% of control activity while activity in the fetal
brain was 65%. There was significant fi inhibition of AChE activity in all
tissues taken from animals at the mid-dose (20 mg kg−1 bw) 1 hr after admin-
istration. The effects were still apparent in blood and liver from dams and
fetuses after 5 hr. The lowest dose (2 mg kg−1 bw) produced a significant
fi
decrease in AChE activity in several tissues after 1 and 5 hr with no appar-
ent effect after 24 hr. AChE determinations were made using the method
of Ellman et al. (1961).

Propoxur
A nontoxic dose (5 mg kg−1 bw) of propoxur is potentiated by IP pretreat-
ment of rats with 1.0 mg kg−1 bw iso-OMPA (Gupta and Kadel 1990). CaE
activity was markedly reduced, indicating that propoxur toxicity is enhanced
by the inhibition of this hydrolase. Carbamylation of AChE produces an
accumulation of ACh and the picture of muscarinic and nicotinic poisoning.
Spontaneous hydrolysis of the carbamate–ChE complex occurs in vivo,
leading to the disappearance of clinical effects within 24 hr. Penetration
of the blood–brain barrier by the carbamates is insignificant;
fi for this
reason, few central nervous system (CNS) symptoms occur (Ellenhorn and
Barceloux 1988).

Thiodicarb
Thiodicarb is hydrolyzed to methomyl in animals and humans. The antiChE
properties of thiodicarb are related to the rate that methomyl is formed in
metabolizing tissues and made available to tissue ChEs. In a rat dose–
response study, a dermal dose of 33.3 mg kg−1 bw (322 μg cm−2, 25 cm2 treated
skin) resulted in 50% red cell ChE inhibition in 24 hr (Knaak and Wilson
98 J.B. Knaak et al.

1985). Carbaryl at dermal dose levels of 417 mg kg−1 bw (4,000 μg cm−2, 25 cm2
treated) produced no red cell ChE inhibition after 24 hr. An automated
Ellman procedure using a Technicon AutoAnalyzer II Continuous-Flow
Analytical system was used to analyze blood for AChE inhibition (Knaak
et al. 1978, 1980). No information is available concerning AChE reactiva-
tion using the automated Ellman procedure.

V. Target Enzymes, the ChEs and CaEs

AChE terminates nerve impulses by catalyzing the hydrolysis of the neu-
rotransmitter ACh. Carbamate insecticides control insects by the direct
inhibition of AChE in their nervous systems, which eventually leads to respi-
ratory failure and death. A somewhat similar set of events occurs in humans
exposed to toxic amounts of these insecticides, except the environmental
dose received by exposed individuals is usually lower per kg bw, and recov-
ery generally takes place before respiratory failure or other problems occur.
To model (i.e., PBPK/PD model) these events in animals and humans,
the following information should be available: (1) inhibited enzymes (i.e.,
AChE and BChE), (2) preferred substrate, (3) location of enzymes in
fic content (μmol kg−1 tissue), and (4) their bimolecular
tissues, (4) specifi
inhibition rates.
Nigg and Knaak (2000) reviewed the status of the ChEs as part of a
review on blood ChEs as human biomarkers of OP pesticide exposure.
Carbamate insecticides have been implicated in cases of food (Hunter
et al. 1997; Wilson et al. 1989) and applicator poisoning (Knaak, personal
communication). The food poisoning resulted from the application of aldi-
carb (unregistered use) on watermelons in California, while the applicator
poisoning involved clothing contaminated with both methomyl and para-
thion in Imperial County, CA, during the 1970s. The rapid recovery of ChE
inhibition by carbamate insecticides has prevented efforts to clearly relate
reported illnesses to carbamate exposures. Blood samples must be collected
as soon as the illnesses are reported and appropriately analyzed for ChE
inhibition (Wilson et al. 1997, 2002). Knowledge of the location, function
and structure of the ChEs is needed for understanding their physiological
importance and for properly modeling inhibition and recovery.
The CaEs in liver and other tissues are also inhibited by carbamate
insecticides. The B-esterases (CaEs) hydrolyze carboxylesters of natural
products and drugs, and function as scavengers in the removal of carba-
mates and OPs. These enzymes are also believed to hydrolyze carbamates,
rendering them useless as insecticides.

A. Structure, Multiple Forms, and Distribution

AChE
The structure of AChE from the eel Torpedo californica was reported by
Sussman et al. (1991). The monomer is an α/β protein that contains 537
 Parameters for Carbamate Models 99

amino acids and structurally is a 12-stranded mixed β-sheet surrounded by

14 α-helices. Amino acid sequences are known only for Torpedo AChE,
Drosophila AChE, 85% of fetal bovine AChE, and human BChE. The
postulated “anionic site” of AChE that binds the quaternary ammonium
ion of ACh is located in a gorge represented by 14 aromatic residues.
Charges in the anionic site and active site are believed to stabilize the Ch
group. The esteratic or active site serine is at amino acid 200 for red blood
cell (RBC) AChE, and at amino acid 198 for plasma BChE (Lockridge
et al. 1987; Sutton et al. 1991). The binding sites are represented in Fig. 8
by Sussman et al. (1991). The esteratic site is embedded in a gorge about
20 Å long that reaches halfway into the three-dimensional structure of the
protein and is responsible for the hydrolysis of ACh.
AChE exists as mono-, di-, and tetramers of catalytic subunits (G1, G2,
G4) with each unit containing active sites. The most complex form, A12,
has 12 subunits. The forms are either hydrophilic (soluble) or tightly bound
to a phospholipid membrane. The molecular differences in AChE and
BChE were reviewed by Chatonnet and Lockridge (1989) and are given
in Table 7. Asymmetric or immobilized forms of AChE are found only in
peripheral nerves and muscles of vertebrates. Membrane-bound G4 AChE
is found in mammalian brain, while membrane-bound G2 AChE is found
in erythrocytes.

Fig. 8. Schematic representation of the binding sites of AChE based upon previous
kinetic, spectroscopic, and chemical modification
fi studies: ES, esteratic site; AS,
anionic substrate-binding site; ACS, active site-selective aromatic cation-binding
site; PAS, peripheral anionic binding site or sites. The hatched areas represent puta-
tive hydrophobic binding regions. The ACh molecule is shown spanning the este-
ratic and anionic sites of the catalytic center. Imidazole and hydroxyl side chains of
His and Ser are shown with the esteratic site. Within the anionic site, (COO-)n
represents six to nine putative negative charges. (Redrawn from Sussman JL et al.
1991 with permission from AAAS).
100 J.B. Knaak et al.

Table 7. Molecular Isoforms of AChE and BChE.

Hydrophilic, water-soluble forms secreted into body fluids

fl

AChE
G1a G2a G4a A12b
Degradation Degradation Secreted by adrenal
product of product of G4 gland; peripheral nerve
G4 cells, found in plasma,
spinal flfluid
BChE
G1 G2 G4 A12
Degradation Degradation Human plasma BChE,
product of product of G4 95% of activity in
G4 plasma
Immobilized forms, asymmetrical
AChE
G1 G2 G4 A12
Torpedo californica
muscle of primitive
vertebrate
BChE
G1 G2 G4 A12
Muscle of
mammals and birds
Amphiphilic globular forms (membrane bound anchored to
phospholipids bilayers)
AChE
G1 G2 G4 A12
Erythrocyte, Mammalian brain, 20-
glycolipid anchor- kDa anchor containing
PIc, cleaved by fatty acids
protease
BChE
G1 G2 G4 A12
Heart of Torpedo Detergent-soluble form
marmorata and reported in mammalian
superior cervical brain
ganglion of rat

Source: Chatonnet and Lockridge (1989).

a
Globular forms: G1, G2, and G4 contain one, two, or four subunits. G4 tetramer is an
association of two dimmers linked by disulfi
fide bonds; dimmers and monomers appear to be
degradation products of tetramers.
b
A12 contains three tetramers.
c
PI, phosphatidylinositol.
 Parameters for Carbamate Models 101

Genomic DNA analysis and genetic lineage studies from different species
suggest that all molecular forms of AChE in higher vertebrates are a
product of a single gene (Maulet et al. 1990; Randall et al. 1987; Rotundo
1988; Sikorav et al. 1987), while structural diversity (i.e., G1, G2, G4, A12) is
a product of posttranscriptional and translational events.

BChE
BChE is present in regions of the brain in positions not related to AChE,
such as capillary endothelial cells, in glial cells, and in neurons. Human
BChE is synthesized in the brain. Plasma BChE is a 24% carbohydrate
sialoglycoprotein synthesized in the liver (Khoury et al. 1987). In plasma,
the products of the BChE gene (BCHE) exist in multiple molecular forms.
Three main forms can be recognized by gel electrophoresis: they are des-
ignated in order of decreasing mobility as G1, G2, and G4. The G4 form
represents about 95% of the activity. The forms represent globular (G)
monomeric, dimeric, and tetrameric forms of the enzyme. G4 is a highly
stable enzyme (Masson and Goasdoue 1986). Earlier studies show that a
part of G4 can dissociate spontaneously into G1 and G2 (La Motta et al.
1965; Masson 1979).
Studies involving genomic blots and restriction maps indicate that there
is only one human BChE gene (Arpagaus et al. 1990; Lockridge and La Du
1991) located on the q arm of chromosome 3 (Sparkes et al. 1984; Zakut
et al. 1989). The enzyme shows a complex genetic and molecular polymor-
phism that appears to be mainly the result of posttranslational modifica- fi
tions of the gene products. Genomic blots provide strong evidence for one
gene in the monkey, dog, rat, mouse, guinea pig, cow, sheep, pig, rabbit,
and chicken. The complete amino acid sequence of the usual form of BChE
(Lockridge et al. 1987) was used to find the cDNA clone (McTiernan et al.
1987), while cDNA was used to find fi the gene (Arpagaus et al. 1990).
No genetic variants are known for human RBC AChE. About 10 genetic
variants are known for plasma BChE (Lockridge 1990). The human BChE
gene contains four exons. Exons 2, 3, and 4 contain sequences for the
globular, tetrameric BChE. Exon 2 encodes 83% of the mature protein,
from the N-terminal of the protein, Glu 1 to Gly 478. Exon 2 encodes the
active site serine, Ser 198, the putative anionic site component, Asp 70, the
potential active site histidine, His 438, seven of the total of nine carbo-
hydrate chains per subunit, and two disulfide fi loops, Cys 65–92 and Cys
252–263.
Five genetic variants of human BCHE have been mapped to exon 2.
Variants H, J, and K have 90%, 66%, and 33% lower activity, respectively.
Atypical is the dibucaine resistant variant, whereas Silent-1 has no activity.
The atypical variant has a reduced Km for all positively charged substrates
and inhibitors but a normal kcat. Atypical BChE has a glycine in position
70 in place of aspartic acid.
102 J.B. Knaak et al.

The J variant has been mapped to exon 3. Antibody assays indicate that
the amount of BChE protein is reduced. In the J variant, the charged amino
acid, Glu 497, is replaced by the hydrophobic residue valine.
The K variant has been mapped to exon 4. Antibody assays (Rubinstein
1978) indicate a reduced amount of serum protein. Glycine is substituted
for Ala at position 539. The K variant is the most common genetic variant,
occurring in homozygous form in 1.7% of the Caucasian population and in
heterozygous form in 22.6% of the population. The form is not associated
with any abnormal drug response.

CaE
CaEs are members of the α/β hydrolase family of enzymes, which contain
a catalytic triad consisting of a nucleophile (Ser-221), a base (His-467), and
an orientating acid (Glu-353) and are localized in the endoplasmic reticu-
lum of many tissues (Hosokawa et al. 1987, 1990; Satoh and Hosokawa
1998). Mammalian liver CaEs belong to a family of proteins encoded by
multiple genes. The isozymes were initially classified fi by their substrate
specificities
fi and pI (isoionic point). The isozymes are now classifi fied into
four main CES groups (carboxylesterase, EC 3.1.1.1) and several subgroups.
Two major human liver isozymes, hCE-1 and hCE-2, belong to classes
CES1 and CES2. By Northern blot, a single band of approximately 2.1
kilobases (kb) was seen for hCE-1 (Riddles et al. 1991), and three bands
of approximately 2, 3 and 4.2-kb were seen with hCE-2 (Schwer et al. 1997).
The intensities of the 2.1-kb band were liver >> heart > stomach > tes-
tis = kidney = spleen > colon > other tissues. In the case of hCE-2, the 2-kb
band was located in liver > colon > small intestine > heart, the 3-kb band in
liver > small intestine > colon > heart, and the 4.2-kb band in brain, testis,
and kidney only. Analysis of substrate structure versus effi ficiency for the
ester or carbamate substrates reveals that the two CaEs recognize different
structural features of the substrate (i.e., acid, alcohol, etc.). The catalytic
mechanism involves the formation of an acyl-enzyme on an active serine.
Knowledge of the substrate structure–activity relationships and the tissue
distribution of CE (CaE = CE) are critical for predicting the metabolism
and pharmacokinetics of pesticides in humans.

B. AChE, BChE, and CaE Substrate Selectivities

(Activity in Crude Tissue Preparations, Concentrations in Tissues,
and Effects of Tetramer on Active Sites and Purifi
fication)
Mutation studies indicate that BChE catalyses ACh hydrolysis as effificiently
as AChE, when six of the active site gorge aromatic residues, with human
AChE numbers (HuAChE) 72, 124, 286, 295, 297, and 337, are replaced by
aliphatic amino acids (Cygler et al. 1993; Harel et al. 1992). On the other
hand, AChE and BChE exhibit distinct substrate and inhibitor selectivities,
and some of the differences are associated with the nature of the amino
 Parameters for Carbamate Models 103

acids at each of these positions. Early work indicated that the main func-
tional difference between the AChE and BChE active sites is related to the
structure of the acyl pocket, where residues corresponding to Phe295 (288)
and Phe297 (290) are replaced by Leu and Val, respectively. BChE is more
reactive than AChE toward bulky substrates such as butyrylcholine (BCh)
(where kcat/Km = 22 × 108 vs. 0.3 × 108 M−1 min−1) or OP inhibitors such as
diisopropyl phosphofl fluoridate (DFP, CAS no. 55-91-4) (where ki = 1.0 × 105
vs. 1.66 × 10 M min−1) and paraoxon (CAS no. 311-45-5) (Kaplan et al.
7 −1

2001). The primary biological role of human CaE (hCE1) is xenobiotic

metabolism. The enzyme is capable of cleaving ester, amide, and thioester
linkages in a wide variety of compounds. Single site mutations in the active
site of hCE1 convert the enzyme into an effi ficient OP insecticide hydrolase
that is able to detoxify nerve agents (Redinbo et al. 2003).

Determination of AChE, BChE and CaE in crude tissue preparations

Maxwell et al. (1987) determined the esteratic binding sites of AChE,
BChE, and CaE in tissues of control rats for purposes of determing the
relationship between inhibition and detoxification.
fi The methods that were
used to obtained information on enzyme activity and turnover rates are
important because the results (esteratic sites) were used in PBPK/PD
models developed by Gearhart et al. (1990, 1994) and Timchalk et al. (2002)
to predict inhibition. The methods used by Maxwell et al. (1987) to obtain
enzyme activity are briefly
fl described below.
Rats were exsanguinated and perfused via cardiac puncture with 150 mL
saline before tissue harvesting and homogenization. Brain tissue was
homogenized in 9 vol ice-cold saline containing 1% (v/v) Triton X-100 using
a Potter-Elvehjem tissue homogenizer. Spleen, lung, kidney, and liver were
homogenized in the same manner except that 4 vol saline with 1% (v/v)
Triton X-100 was used. The diaphragm, heart, intestine, and skeletal muscle
were homogenized in 4 vol saline by three 10-sec pulses of a Polytron
homogenizer (Brinkman Instruments, Westbury, NY) at a speed setting of
eight. Triton X-100 was added equal to 1% of the final volume and gently
rotated for 10 min. All homogenates were centrifuged at 15,000 g for 10 min
in a refrigerated centrifuge (4°–6°C), and the supernatants were stored
frozen at −20°C before ChE analysis.
Assays of ChE were performed (Groff et al. 1976) using acetylthiocho-
line (ATCh) as substrate. Estimates of the total ChE and AChE was accom-
plished by ChE determinations in the absence and presence of iso-OMPA,
a specifi
fic inhibitor of BChE (Austin and Berry 1953). Inhibition of BChE
was performed by the method of Michalek et al. (1983) using 10 μM iso-
OMPA for all tissues except plasma, where 100 μM iso-OMPA was used
(Grubic et al. 1981).
CaE activity was measured using an automatic pH stat (Radiometer
America, Cleveland, OH) with 0.2% tributyrin in saline as substrate at pH
104 J.B. Knaak et al.

Table 8. Enzyme Activities in Control Rat Tissues.c

Enzyme activity (μmol min−1 g−1 tissue)

Cholinesterase Detoxifying enzymes

Tissue Total ChEa AChEa BChEb CaEa

Brain 8.12 ± 1.42 7.34 ± 1.25 0.78 1.0 ± 0.2

Lung 1.82 ± 0.25 0.38 ± 0.06 1.44 23.4 ± 1.9
Spleen 1.86 ± 0.18 1.00 ± 0.15 0.86 5.4 ± 0.3
Muscle 1.66 ± 0.09 1.29 ± 0.14 0.37 4.1 ± 0.5
Diaphragm 1.73 ± 0.16 1.29 ± 0.18 0.44 5.3 ± 0.4
Intestine 6.76 ± 0.80 0.71 ± 0.3 6.05 385.7 ± 59.6
Kidney 0.26 ± 0.04 0.09 ± 0.03 0.17 29.8 ± 6.3
Heart 5.65 ± 0.59 0.84 ± 0.07 4.81 3.6 ± 0.5
Liver 0.67 ± 0.06 0.17 ± 0.04 0.5 32.4 ± 4.2
Plasma 0.48 ± 0.07 0.22 ± 0.02 0.26 7.6 ± 1.7
a
Total ChE, AChE and CaE activities are expressed as x ± standard deviation, n = 9.
b
BChE = total ChE − AChE.
c
Adapted from Maxwell et al. (1987).

8.0. The reaction was started by the addition of 2–200 μL homogenate. CaE
activity was determined by titrating liberated acid with 0.01 N NaOH. The
enzyme activities (μmol min−1 g−1 tissue) in control rat tissues as determined
by Maxwell et al. (1987) are given in Table 8. BChE activity was determined
by subtracting AChE activity from total ChE activity.

Active sites, a measure of B-esterase concentrations in tissue

The number of active sites of AChE, BChE, and CaE (i.e., B-esterase
concentrations in tissues) were used in PBPK/PD models by Gearhart et
al. (1990, 1994), Knaak et al. (2002), and Timchalk et al. (2002) to describe
inhibition by DFP/paraoxon, paraoxon/isofenphos-oxon, and chlorpyrifos-
oxon, respectively. The numbers of active sites (enzyme concentration g−1
tissue) were estimated by Maxwell et al. (1987) in plasma, liver, brain, and
other regions by dividing esterase activity (μmol min−1 g−1 tissue) in control
rat tissues (see Table 8) by their turnover rates (substrate hydrolyzed/min/
active site). The turnover rates (expressed in hr−1) are shown as AChE,
1.81 × 105 from Wang and Murphy (1982); BChE, 6.10 × 104 from Main
et al. (1972); and CaE, 1.81 × 103 from Ikeda et al. (1977) in Tables 9, 10,
and 11, respectively.
In practice, Knaak et al. (2002) used the esterase binding sites from
Maxwell et al. (1987) in μmol B-esterase L−1 tissue, bimolecular inhibition
rate constant (μM−1 hr−1), concentration of carbamate in tissue (μmol L−1),
and volume of tissue (L) in their PBPK/PD model to determine inhibition
levels, as shown in Eq. 23 for the brain.
Table 9. Estimated AChE Available for Inhibition/Detoxification
fi of Carbamate Pesticides.a

Enzyme activities Esterase binding sites based on turnover rates

Tissue μmol min−1 g−1 μmol hr−1 kg−1 μmol kg−1c μmol kg−1d μmol kg−1e μmol kg−1f Mean μmol kg−1 SDb μmol kg−1

Brain 7.34 4.40 × 105 3.76 × 10−2 2.86 × 10−2 4.89 × 10−2 5.65 × 10−2 4.29 × 10−2 1.23 × 10−2
Lung 0.38 2.28 × 104 1.95 × 10−3 1.48 × 10−3 2.53 × 10−3 2.92 × 10−3 2.22 × 10−3 6.36 × 10−4
Spleen 1.0 6.00 × 104 5.13 × 10−3 3.89 × 10−3 6.67 × 10−3 7.69 × 10−3 5.85 × 10−3 1.67 × 10−3
Muscle 1.29 7.74 × 104 6.62 × 10−3 5.02 × 10−3 8.60 × 10−3 9.92 × 10−3 7.54 × 10−3 2.16 × 10−3
Diaphragm 1.29 7.74 × 104 6.62 × 10−3 5.02 × 10−3 8.60 × 10−3 9.92 × 10−3 7.54 × 10−3 2.16 × 10−3
Intestine 0.71 4.26 × 104 3.64 × 10−3 2.76 × 10−3 4.73 × 10−3 5.46 × 10−3 4.15 × 10−3 1.19 × 10−3
Kidney 0.09 5.40 × 103 4.62 × 10−4 3.50 × 10−4 6.00 × 10−4 6.92 × 10−4 5.26 × 10−4 1.51 × 10−4
Heart 0.84 5.04 × 104 4.31 × 10−3 3.27 × 10−3 5.60 × 10−3 6.46 × 10−3 4.91 × 10−3 1.41 × 10−3
Liver 0.17 1.02 × 104 8.72 × 10−4 6.62 × 10−4 1.13 × 10−3 1.31 × 10−4 9.94 × 10−4 2.85 × 10−4
Plasma 0.22 1.32 × 104 1.13 × 10−3 8.57 × 10−4 1.47 × 10−3 1.69 × 10−3 1.29 × 10−3 3.68 × 10−4
Blood 1.32 × 104 1.128 × 10−3
a
Turnover rates (substrate hydrolyzed hr−1 active site−1) used for calculations.
Parameters for Carbamate Models

Enzyme activities (μmol min−1 g−1 tissue) for calculations of binding sites were taken from Table 8.
Esterase binding sites = (enzyme activity)/turnover rate.
b
Coeffi
ficient of variation = 28.64%.
c
Turnover: 1.17 × 107 hr−1; Wang and Murphy (1982).
d
Turnover: 1.54 × 107 hr−1; Torpedo, Selwood et al. (1993).
e
Turnover: 9.00 × 106 hr−1; Torpedo/recombinant, Radic et al. (1992).
f
Turnover: 7.80 × 106 hr−1; Mouse/wild type, Radic et al. (1992).
105
106 J.B. Knaak et al.

Table 10. Estimated BChE Available for Inhibition/Detoxifi

fication of Carbamate
Pesticides.a

Esterase binding sites based on

Enzyme activity turnover rates

Tissue μmol min−1 g−1 μmol hr−1 kg−1 μmol kg−1 b μmol kg−1c μmol kg−1d

Brain 0.78 4.68 × 104 1.28 × 10−2 5.94 × 10−3 7.09 × 10−3
Lung 1.44 8.64 × 104 2.36 × 10−2 1.10 × 10−2 1.31 × 10−2
Spleen 0.86 5.16 × 104 1.41 × 10−2 6.54 × 10−3 7.82 × 10−3
Muscle 0.37 2.22 × 104 6.07 × 10−3 2.82 × 10−3 3.36 × 10−3
Diaphragm 0.44 2.64 × 104 7.21 × 10−3 3.35 × 10−3 4.00 × 10−3
Intestine 6.05 3.63 × 105 9.92 × 10−2 4.60 × 10−2 5.50 × 10−2
Kidney 0.17 1.02 × 104 2.79 × 10−3 1.29 × 10−3 1.55 × 10−3
Heart 4.81 2.89 × 105 7.89 × 10−2 3.66 × 10−2 4.37 × 10−2
Liver 0.5 3.00 × 104 8.20 × 10−3 3.81 × 10−3 4.55 × 10−3
Plasma 0.26 1.56 × 104 4.26 × 10−3 1.98 × 10−3 2.36 × 10−3
a
See Table 9 for calculation of enzyme activity and esterase binding sites based on turnover
rates.
b
Turnover: 3.66 × 106 hr−1; Main et al. (1972).
c
Turnover: 7.88 × 106 hr−1; Torpedo, Golicnik et al. (2002) .
d
Turnover: 6.60 × 106 hr−1; Kaplan et al. (2001).

Table 11. Estimated CaE Available for Inhibition/Detoxifi

fication of Carbamate
Pesticides.a

Esterase binding sites

Enzyme activity based on turnover rates

Tissue μmol min−1 g−1 μmol hr−1 kg−1 μmol kg−1b μmol kg−1c

Brain 1.0 6.00 × 104 5.52 × 10−1 1.68 × 100

Lung 23.4 1.40 × 106 1.29 × 10−1 3.93 × 10+1
Spleen 5.4 3.24 × 105 2.98 × 100 9.09 × 100
Muscle 4.1 2.46 × 105 2.27 × 100 6.90 × 100
Diaphragm 5.3 3.18 × 105 2.93 × 100 8.92 × 100
Intestine 385.7 2.31 × 107 2.13 × 102 6.48 × 10+2
Kidney 29.8 1.79 × 106 1.65 × 10+1 5.02 × 10+1
Heart 3.6 2.16 × 105 1.99 × 100 6.06 × 100
Liver 32.4 1.94 × 106 1.79 × 10+1 5.44 × 10+1
Plasma 7.6 4.56 × 105 4.20 × 100 1.29 × 10+1
a
See Table 9 for calculation of enzyme activity and esterase binding sites based on turnover
rates.
b
Turnover: 1.09 × 105 hr−1; Ikeda et al. (1977).
c
Turnover: 3.56 × 104 hr−1; Stok et al. (2004).
 Parameters for Carbamate Models 107

Equation for the inhibition of blood AChE by carbamates:

dAiAChEB/dt = (KiAChEBCAChEBCcarbamateVB) (μmol hr−1) (23)
where VB = volume of blood (L),
KiAChEB = AChE bimolecular inhibition rate constant (μM−1 h−1),
CAChEB = concentration of free AChE in blood (μmol L−1), and
Ccarbamate = concentration of carbamate in blood (μmol L−1).

In all three PBPK/PD models [Gearhart et al. (1990, 1994), Knaak et al.
(2002), and Timchalk et al. (2002)], the active site data from the rat were
used in human models. The PD portion of these models is highly dependent
on these values. New estimates are needed to validate OP and carbamate
pesticide PBPK/PD models. Additional turnover numbers, kcat, (Radic
et al. 1992; Pyror et al. 1992; Selwood et al. 1993; Radic et al. 1993; Kaplan
et al. 2001) for the hydrolysis of thiocholinesters by AChE and BChE were
found and used to obtain new estimates of μmol active sites per kg tissue.
One kcat value was found for CaE (Stok et al. 2004). The results are shown
in Tables 9 and 10.
A 28.6% coefficient
fi of variation for AChE was determined for the
various tissues based on the new turnover numbers in Table 9. This varia-
tion suggests that the output (inhibition) from PBPK/PD models may vary
by at least this percentage. The source of enzymes and methods for deter-
mining turnover numbers was found to vary between studies.
In a study by Pryor et al. (1992), the active sites of electric eel (EE)
AChE were measured by titration with umbelliferyl diethyl phosphate
(UDP) (CAS no. 299-45-6). The release of umbelliferone (CAS no. 93-35-
6) from the reaction with EE-AChE was monitored by fluorescence
fl spec-
troscopy. The time course of fluorescence emission increased following the
addition of AChE to the titration solution. The data (fluorescence
fl vs. time
in sec) fifit the following first-order equation:
F = (F0 − Finf)e−kt + Finff (24)
where F, F0, and Finff are the emission intensities of time t, 0, and infi
finity,
respectively; k is the first-order rate constant (3.1 × 107 hr−1). The enzyme
concentration was determined from the value of Finff − Fb (Fb, background
fluorescence) using the calibration plot. Division of k by the concent-
fl
ration of UDP gave a second-order bimolecular rate constant of
2.77 × 1011 M−1 hr −1. This rate constant is similar to the rate constant
(1.71 × 1011 M−1 hr −1) determined by Kaplan et al. (2001) for human AChE
and ATCh in which the hydrolytic parameters (Km, kcat, and kcat/Km) of
human recombinant AChE and BChE against ATCh and BTCh (acetyl
and butyrylthiocholine) were assayed according to Ellman et al. (1961)
and monitored using a Thermomax microplate reader (Molecular
Devices, Sunnyvale, CA). Enzyme concentration was determined by
ELISA (enzyme-linked immunosorbent assay) (Shafferman et al. 1992)
108 J.B. Knaak et al.

and by active site titration (Velan et al. 1991) using the PsCs-soman
stereomer (CAS no. 24753-16-0). Human AChE displayed a 50-fold
difference in reactivity for ATCh over BTCh with kcat being 2.4 × 107 hr −1
and 4.8 × 105 hr −1, respectively. Human BChE was more reactive against
BTCh than AChE (kcat = 6.6 × 106 hr −1 and 4.8 × 105 hr −1, respectively).

Effects of tetramer and molecular forms on active site measurements

The tetramer is the most important form for AChE in physiological condi-
tions (i.e., neuromuscular junction and the nervous system). A study by
Zhang et al. (2005) involving diffusion of ACh to the active sites of the
tetramer show that the reaction rates for three mouse AChE tetramers
differ for individual active sites in the tetramer. Depending upon salt con-
centration, the rates for active sites in the tetramer are equivalent to 67%–
75% of the rate for the monomer. Measurements made on AChE activity
in tissues may underestimate the amount of available enzyme for carrying
out ACh hydrolysis. In an earlier study by Gordon et al. (1978), the turn-
over numbers of different forms (18 S, 14 S, and 8 S asymmetrical forms
and the 11 S form) from electric eel were measured using an active phos-
phorylating inhibitor as a titrating agent. The 8 S and 11 S forms were
approximately 35% more active than the 18 S and 14 S forms.

Purification
fi of AChE, BChE, and CaE
Purified/recombinant
fi enzymes are needed for studying the kinetics of their
catalytic reactions. Rosenberry and Scoggin (1984) purifiedfi human eryth-
rocyte AChE on an acridinium resin affinityfi column. Five mg of enzyme
were obtained from 10 L of outdated erythrocytes. The purifi fied enzyme had
a specifific activity of 5000–5800 units mg−1 protein (unit = 1.0 μmol min−1;
5.0–5.8 mmol min−1 mg−1 protein) and was free of polypeptide contaminants
by gel electrophoresis. In detergents, the isolated enzyme corresponded to
a disulfide-linked
fi dimer (G2) that was converted to 75-kDa subunit mono-
mers (G1) by reduction with dithiothreitol. G1 contained 1.7 mol free sulf-
hydryl groups mol −1 subunit. This study provided strong evidence that
erythrocyte AChE is an amphipathic protein.
Affifinity chromatography, of mutant mouse AChE in serum free media,
with trimethyl (m-aminophenyl) ammonium linked through a long tether
arm to Sepharose CL-4B resin (Sigma, St. Louis, MO) permitted a one-step
purification
fi of AChE, both mutant and wild-type in amounts between 5
and 25 mg, as previously described. Purity was assessed by SDS-PAGE
(sodium dodecyl sulfate-polyarcylamide gel electrophoresis) and by com-
parisons of specific
fi activity with absorbance at 280 nm to measure protein
concentration (ε280 = 1.14 × 105 M−1 cm−1) (Boyd et al. 2004). The catalytic
activity of each unlabeled and labeled mutant was measured with the
Ellman assay. Km and the dissociation constant of a ternary complex result-
ing in substrate inhibition or activation, Kss, were kinetically evaluated.
 Parameters for Carbamate Models 109

The x-intercept of a plot of the residual catalytic activity versus the con-
centration of the irreversible inhibitor 7-(methylethoxyphosphinyloxy)-1-
methylquinolinium iodide (MEPQ) (CAS no. 95230-44-7) yielded the
enzyme concentration, and, in turn, kcat. No data were supplied for kcat, Km,
and Kss.
Two types of CaE (esterase I and II) were purified fi from rat liver micro-
somes after 90% of the triacylglycerol lipase activity was released by heparin
treatment. Purifification of the residual fraction involved diethylaminoethyl
(DEAE)-cellulose chromatography, hydroxyapatite chromatography,
isoelectric focusing, and Sephadex G-200 chromatography. The final fi
preparation of esterases I and II, which were purified fi 70- and 140 fold,
respectively, gave single protein bands on polyacrylamide gel and sodium
dodecyl sulfate-gel electrophoresis. The molecular weights of esterases I
and II were calculated to be about 70,000 and 160,000 by gel fi filtration on
Sephadex G-200. Esterase I differed immunologically from esterase II, and
it was found to constitute about 30% of the total esterase activity in the
microsomal fraction; esterase II constituted 50%–60% (Ikeda et al. 1977).
Sanghani et al. (2004) purified
fi human isozymes hCE1 and hCE2 from
class 1 and 2 CaEs, respectively, using a modification
fi of the method of
Humerickhouse et al. (2000). Briefly,fl 85 g human liver was first homoge-
nized and centrifuged at 35,000 g for 40 min. The supernatant was added
to DE52 resin, the resin washed, and the CaEs eluted from the resin and
further purified
fi on a 50-mL concanavalin A column. Human isozymes
hCE1 and hCE2 were separated on a preparative nondenaturing gel elec-
trophoresis column in a Bio-Rad Prep Cell model 491 (Bio-Rad, Hercu-
les, CA). Purified
fi hCE1 and hCE2 exhibited specifi fic activities of about 7
and 140 units mg−1, respectively, using methylumbelliferyl acetate as sub-
strate. A unit is defi fined as 1.0 μmol substrate hydrolyzed min−1 mg−1
enzyme.

VI. Acylation and Decarbamylation of ChEs and CaEs

A. Hydrolysis of Substrates
The hydrolysis of ACh and BCh by ChE is extremely rapid, making it dif-
ficult to measure the rate of hydrolysis. To handle this problem, activity
fi
measurements are made under steady-state conditions or after the partial
inhibition of ChE by an irreversible inhibitor. In most preparations the
actual molar concentrations of enzyme are unknown and the amount of
fic activity (i.e., units mg−1
enzyme can be expressed only in terms of its specifi
−1 −1
protein; units μmol enzyme; Vmax mg protein). Turnover number or rate,
kp, k2, or kcat, is defined
fi as follows:

k −1
Vmax pmoles(substrate product)min 1 kg
k cat (min 1 ) = = (25)
[E]t μmolles(enzyme)kg −1
110 J.B. Knaak et al.

where total enzyme is given as

[E]t = [E] + [ES] (26)
The reciprocal of kcat gives the time required for a single catalytic cycle
(Segel 1993). The following section is a review of the kinetics of the
reaction.

Acylation/hydrolysis, AChE, BChE, and CaE

AChE (EH) combines with acetylcholine (AB) resulting in a Michaelis–
Menten complex (EHAB) that is converted to an acylated enzyme (EA)
and a leaving group (BH) (Aldridge and Reiner 1972). The acylated enzyme
decomposes into an enzyme product as indicated below:
EH AB ← → EHAB
k +1 k k 3
EA BH EH AOH (27)
k −1
+ H 2O
where k+1, k−1, k+2 and k+3 are the velocity (sec−1) of the equilibrium reac-
tions. BH and AOH are the products of substrate hydrolysis, where BH is
Ch and AOH is acetic acid, respectively. The EA formed from ACh hydro-
lyzes very quickly (t1/2 ≤ 2.3 × 10−6 min) (Wilson 1951a,b; Cohen et al. 1955).
These rates are initially measured under steady state conditions when the
concentrations of all species (EH, EHAB, and EA) are constant. The
steady-state kinetic parameters (Pryor et al. 1992) are given by the follow-
ing equations:
k +1 k +2 [E]T
Vmax /K
Km (k cat [E]T ) K m = (28)
k −1 k + 2
k +2 k +3 [E]T
Vmax k cat [E]T = (29)
k +2 k + 3
where [E]T is the total enzyme.
The second-order rate constant, kcat/Km, for ACh and ATCh hydrolysis
is ∼108 M−1sec−1 (3.6 × 1011 M−1 hr−1) and is diffusion controlled. The turnover
number, kcat, is ∼3.6 × 107 hr−1. Pryor et al. (1992) determined the kinetic
parameters Vmax and Km by nonlinear least-squares fitting of Vi and [S] to
the Michaelis–Menten equation, where Vi and [S] are the initial velocity
and substrate concentration, respectively:
V[S]
Vi = (30)
K [S]
Values of kcat and kcat/Km were calculated by dividing Vmax and Vmax/Km,
respectively, by the enzyme concentration (mass of 70,000 per active site).
Initial velocities were measured by linear least-squares fitting of time
courses for ≤5% of total substrate turnover using the assay of Ellman et al.
(1961).
 Parameters for Carbamate Models 111

Kinetic activity of AChE, BChE, and CaE

Pryor et al. (1992) determined the kinetic constants (kcat, kcat/Km) for the
hydrolysis of TCh esters by Electrophorus electricus AChE (Sigma, St.
Louis, MO), purified
fi human red blood cell AChE (Rosenberry and Scoggin
1984), fetal bovine serum AChE (De La Hoz et al. 1986), and Torpedo
californica AChE (Sussman et al. 1988).
In a study by Radic et al. (1993), mouse wild-type and mutant AChE
and BChE activities were measured in 0.1 M Na3PO4 (pH 7.0) at 22°C by
the assay of Ellman et al. (1961) according to Eq. 31:

(31)

where S combines at two different sites on the enzyme, forming two binary
complexes, ES and SE, of which only ES results in substrate hydrolysis.
Values for Km, Kss, and b were calculated using nonlinear computer fi
fitting
according to the following equation:
1 + b[S] KSS ⎞ ⎛ Vmax ⎞
v=⎛ (32)
⎝ 1 + [S] KSS ⎠ ⎝ 1 + Km [S] ⎠

where Kss is a substrate inhibition constant, and b reflects

fl the effi
ficiency
of hydrolysis of the ternary complex, SES, relative to ES (Aldridge and
Reiner 1972; Webb and Johnson 1969). Active sites were quantitated by
titration with 7-(methylethoxyphosphinyloxy)-1-methylquinolinium iodide
(MEPQ), a high-affi finity phosphorylating agent (Levy and Ashani 1986;
Radic et al. 1992). Inhibition by reversible inhibitors was measured at four
to fifive substrate and three inhibitor concentrations. Reciprocal and regres-
sion plots were used to obtain kinetic parameters. Inhibitor and substrate
binding to AChE and BChE was modeled by an energy minimization
docking program (Discover-Insight 2.2.0; Biosym Technologies, San Diego,
CA) on a Silicon Graphics Indigo Elan (Radic et al. 1992).
Stok et al. (2004) determined the kinetic parameters for the hydrolysis
of p-nitrophenyl acetate by rat CaE 6.1. The values of kcat, the turnover
number, and kcat/Km, the second-order rate constant, were determinded
to be 3.5 × 104 hr−1 and 3.09 × 109 M−1 hr−1, respectively. These values are
1000 and 116 times slower than those reported by Pryor et al. (1992) for
AChE and ACh, respectively. The hydrolytic action of AChE on ACh is
considered to be the most rapid reaction recorded. Carbaryl is an inhibitor
of CaE 6.1, with Kd, k2 and k3 values of 35 μM, 108 hr−1, and 39.6 hr−1,
respectively.
112 J.B. Knaak et al.

Complexities of AChE and BChE kinetic activity

The kinetics of BChE catalyzed reactions display a number of complexities
that could reflect
fl microheterogeneity: (1) substrate hydrolysis deviates
from Michaelis–Menten kinetics (Christian and Beasley 1968; Eriksson and
Augustinsson 1979; Kalow 1964); (2) irreversible inhibition does not follow
first-order kinetics (Main 1969b); and (3) kinetics presents a nonlinear
fi
temperature dependence with a break at 18°–21°C in Arrhenius plots
(Ferro and Masson 1987). In addition, where there are four active sites per
tetramer (Eriksson and Augustinsson 1979; Lockridge and La Du 1978),
lower active site numbers have been reported, e.g., two (Muensch et al.
1976).

B. Carbamylation (Inhibition) and Decarbamylation of ChEs and CaEs

Rate of inhibition characterized by second-order rate constant. The
inhibition of AChE by carbamate insecticides occurs by the following
reactions:

(33)

where EH and AB are the enzyme and carbamate, respectively, EHAB

is the enzyme–inhibitor complex, and Ka = k−1/k1, which is now called the
dissociation constant, Kd, is the binding, affi finity constant. Main (1969a)
described a procedure for measuring the binding constant Ka (M), carba-
mylation constant k2 (hr−1), and the overall bimolecular rate constant ki
(M−1 hr−1). The bimolecular rate constant was determined by plotting the
reciprocal of the inhibition rate against the reciprocal of the inhibitor con-
centration. The slopes give ki, and the intercepts of the extrapolated lines
on the ordinate and abscissa give (−Ka−1) and (k2−1).
Saxena et al. (1997) and Ordentlich et al. (1993) measured the inhibition
bimolecular rate constants (ki) for OPs under pseudo-first-order
fi conditions
from the plots of slopes of ln E (enzyme) versus time at different inhibitor
concentrations. Rate constants under second-order conditions were deter-
mined from nonlinear regression of plots of ln [E/(I0 − (E0 − E))] vs. time
(I0, initial inhibitor concentration).

Inhibition of more than one form of enzyme

Main (1969b) observed that the plots of the log velocity against time for
serum ChE and erythrocyte AChE were not linear with the OP tetram
(CAS no. 78-53-5). Similar curving was observed with neostigmine under
conditions of negligible regeneration. Curving was caused by the inhibition
of two or more forms of ChE. The use of a purified fi preparation of eel
AChE and neostigmine resulted in little or no curving until 99% of the
enzyme had been inhibited.
 Parameters for Carbamate Models 113

Inhibitory activity as function of carbamate structure

The application of the Main (1964) equation to inhibition of AChE by
carbamates has been studied in detail by Hastings et al. (1970) and O’Brien
(1968). Fukuto et al. (1967) assumed that these compounds behave as com-
petitive substrates for ChE with high affinities
fi for the enzyme and low
turnover numbers. The Km for the hydrolysis of ACh chloride by bovine
erythrocytes was reported to be 0.29 × 10−3 M (Hetnarski and O’Brien
1975b). This constant is equivalent to the binding of a carbamate to the
enzyme surface as measured by the dissociation constant, Kd. According to
Hetnarski and O’Brien (1973), variations exist in the inhibitory activity of
aryl methylcarbamates and aryl dimethylcarbamates toward AChE with
the latter compounds inhibiting only by complex formation, and not by
complex formation followed by carbamylation, as is the case with aryl
methylcarbamates. Most carbamates, however, give rise to a carbamylated
enzyme. Rapid decarbamylation results in recovered enzyme when the
enzyme is removed from the surplus inhibitor. Hetnarski and O’Brien
(1973) considered recovery “pseudoreversibility” in the sense that the
enzyme recovers, and the carbamate is hydrolyzed in the course of the
reaction. Potency is normally a function of both affinity
fi and innate carba-
mylation activity. The validity of the Kd and k2 estimations depend upon
inhibition following first-order
fi kinetics when inhibitor concentrations are
constant (Iverson and Main 1969).

CaE inhibition by carbamates

The protective effect of CaE against AChE inhibition by carbamates has
not been well characterized as it has been for the OPs. In the case of OPs,
CaE performs the role of a high-affinity/low-capacity
fi detoxifi
fication system
with ki values > 106 M−1 min−1. CaEs appear to be less effective in the case
of carbamates as Stok et al. (2004) determined the ki for carbaryl to be
5.14 × 104 M−1min−1.
Maxwell (1992) measured the activity of lung, plasma, and liver CaE
against 1-naphthyl acetate using the spectrophotometric method of Ecobi-
chon (1970) in the presence of 10−5 M physostigmine to prevent hydrolysis
by BChE. Physostigmine was not required for measurement of purified fi
plasma CaE activity. The ratio of the inhibition rate constants (ki, ratio:
plasma CaE/brain AChE) for paraoxon (5/1) and a number of OPs of mili-
tary significance
fi were plotted against their protective ratios (LD50 of con-
trols/CaE inhibited animals; ratio of 2 for paraoxon). Small ki ratios of less
than 1.0 gave protective ratios of 4 or greater. The larger number of binding
sites in liver suggests that CaE detoxication may be more important in this
tissue than in plasma or the lung.

Decarbamylation of Inhibited Molecular Forms, First-Order Rate Constant

Spontaneous decarbamylation of both plasma and erythrocyte ChE inhib-
ited by carbamates is an established feature. It is commonly accepted that
114 J.B. Knaak et al.

the binding of carbamates to ChE is reversible and that some hydrolysis

of the carbamate occurs on the enzyme (Aldridge and Reiner 1972;
Rotenberg and Almog 1995). Reactivation of aliquots of carbamylated
AChE was followed after extensive dilution of the inhibited enzyme using
the method of Ellman et al. (1961) (Dawson 1994; Dawson and Poretski
1985).

C. Bimolecular Rate Inhibition Constants, ki, for the Ten Carbamates

Kinetic constants for the inhibition of AChE and BChE are shown in
Tables 12 and 13 for the 10 carbamates and their active metabolites (Dawson
1995; Herzsprung et al. 1992; Hetnarski and O’Brien 1975a; Main 1979;
Vilarem et al. 1989; Yu et al. 1972b). Noteworthy is the large amount of
work carried out by Herzsprung et al. (1992). k2 is assumed to be equivalent
for all the carbamates (Goldblum et al. 1981). Kd was calculated using k2
and ki.
Herzsprung et al. (1992) determined the ki for the inhibition of AChE
from electric eel, AChE from bovine erythrocytes, and BChE from human
and horse serum by aldicarb, aldicarb sulfoxide and sulfone, carbofuran,
3-hydroxycarbofuran, carbaryl, methomyl, oxamyl, pirimicarb, propoxur
and 2-hydroxypropoxur using the colorimetric method of Ellman et al.
(1961). According to Herzsprung et al. (1992), the reaction of the carba-
mates with ChE may be considered a pseudo-first-order
fi reaction when the
concentration of enzyme is small compared to the concentration of
inhibitor:

EH IB ki
→ EI BH
(34)
[EH t ] [EH ] exp( k*i [IB] t)

where [IB] = concentration of carbamate in the incubation mixture

[mol L−1], [EH] = concentration of the active enzyme, EI = enzyme inhibi-
tor complex, BH = leaving group, and t = inhibition time.
The bimolecular rate constant ki is given by
ln 2
ki = (35)
t1/ 2 ∗[IB]
The reaction half-time, t1/2, and ki were determined from a plot of log ACt
vs. t. Dawson (1994) calculated the second-order rate constant for carba-
mylation (ki), and the first-order rate constant for decarbamylation (k3) for
carbaryl from the data described by Aldridge and Reiner (1972). Bovine
erythrocyte AChE (soluble preparation), human erythrocyte AChE (mem-
brane-bound), and rat brain AChE (solubilized with Triton X-100) were
used as sources of AChE by Dawson (1994). The kinetic data presented in
Tables 12 and 13 along with other data are briefly fl discussed next for each
carbamate.
 Parameters for Carbamate Models 115

Table 12. Kinetic Constants for the Inhibition of AChE by Carbamate Pesticides
and for Their Reactivation.

Dissociation Carbamylation Decarbamylation Bimolecular

Carbamate/ constant, constant k2, constant k3, rate constant
Metabolite Kd, (M) (min−1) (min−1) ki, (M−1 min−1)

Aldicarba 1.0 × 10−2 146 0.0109g 1.46 × 104

Aldicarbb 1.23 × 10−4 1.72 1.39 × 104
Aldicarbc 5.0 × 104
Aldicarb — — 0.010 5.6 × 105
sulfoxideb
Aldicarb 1.3 × 106
sulfoxidec
Aldicarb — — 0.010 6.0 × 103
sulfoneb
Aldicarb 1.5 × 104
sulfonec
Carbaryla 5.0 × 10−2 20.0 0.0103g 4.0 × 102
Carbaryld 1.3 × 10−5 1.83 1.4 × 105
Carbaryle 0.71 2.5 × 104
Carbarylc 3.3 × 104
Carbofuranf 2.1 × 10−4 21.6 0.010 1.029 × 105
Carbofuranc 1.7 × 106
3-OH 3.5 × 105
Carbofuranc
Formetanateg 0.010
Methiocarba 6.7 × 10−6 1.2 0.010 1.79 × 105
Methiocarbc 1.0 × 105
Methiocarb 0.7 × 105
sulfoxidec
Methiocarb 0.7 × 104
sulfonec
Methomylb 5.40 × 10−6 0.62 1.14 × 105
(Z isomer)
Methomylh 3.305 × 10−5 2.3 0.0113g 6.96 × 104
1.5 × 105
Oxamylb 9.71 × 10−6 1.28 0.010 1.31 × 105
Oxamylc 3.5 × 105
Pirimicarbg 0.010
Propoxura 1.0 × 10−5 1.1 0.010 1.1 × 105
Propoxurd 2 × 10−5 1.47 7.35 × 105
Propoxurc 1.3 × 105
2-OH 0.010 0.8 × 102
Propoxurc
Thiodicarbg 0.010
a
Main (1979), Vilarem et al. (1989), Herzsprung et al. (1992), dHetnarski and O’Brien
b c

(1975a), eDawson (1994), fYu et al. (1972b), gRotenberg and Almog (1995), hDawson (1995).
116 J.B. Knaak et al.

Table 13. Kinetic Constants for the Inhibition of BChE by Carbamate Pesticides.

Dissociation Carbamylation Bimolecular rate

constant Kd, constant k2, constant ki,
Carbamate/metabolite (M) (min−1) (M−1 min−1)

Aldicarba 8.33 × 10−5 2.0 2.4 × 104

Aldicarb sulfoxidea 3.33 × 10−6 2.0 6.0 × 105
Aldicarb sulfonea 8.0 × 10−6 2.0 2.5 × 105
Carbaryla 1.05 × 10−5 2.0 1.9 × 103
Carbofurana 3.33 × 10−6 2.0 6.0 × 105
3-OH Carbofurana 1.82 × 10−4 2.0 1.1 × 104
Formetanateb — — —
Methiocarba 1.54 × 10−4 2.0 1.3 × 104
Methiocarb sulfoxidea 4.65 × 10−5 2.0 4.3 × 104
Methiocarb sulfonea 6.25 × 10−4 2.0 3.2 × 103
Methomyla 5.13 × 10−4 2.0 3.9 × 103
Oxamyla 6.67 × 10−5 2.0 3.0 × 104
Pirimicarbb — — —
Propoxura 2.5 × 10−3 2.0 0.8 × 103
2-OH Propoxura 0.022 2.0 0.9 × 102
Thiodicarbb — — —
a
Herzsprung et al. (1992).
b
Not available.

Aldicarb-AChE and BChE Inhibition

Herzsprung et al. (1992) and Vilarem et al. (1989) determined ki values for
the inhibition of AChE by aldicarb, aldicarb sulfoxide, and aldicarb sulfone
(see Table 12). Aldicarb sulfoxide is more inhibitory than aldicarb or its
sulfone. Main (1979) reported a ki value for aldicarb similar to the ones
reported by Vilarem et al. (1989) and Herzsprung et al. (1992). According
to Herzsprung et al. (1992), aldicarb sulfoxide is more inhibitory toward
BChE than either aldicarb or its sulfone (see Table 13).

Carbaryl-AChE and BChE Inhibition

Hetnarski and O’Brien (1975a) determined Kd, k2, and ki for carbaryl using
bovine erythrocyte AChE and the zero-time method (Hart and O’Brien
1973; O’Brien 1968). The values were 13 × 10−6 M, 1.83 min−1, and
1.408 × 105 M−1 min−1, respectively, as shown in Table 12. Herzsprung et al.
(1992) determined the ki for eel AChE to be 3.3 × 104 M−1 min−1. Similar
values were determined by Dawson (1994) for AChE. Herzsprung et al.
(1992) determined ki values for carbaryl of 1.9 × 103 and 0.7 × 104 M−1 min−1
against human and horse BChE, respectively (see Table 13).
 Parameters for Carbamate Models 117

Carbofuran-AChE and BChE Inhibition

According to Herzsprung et al. (1992), the inhibition ki values for carbofu-
ran and 3-OH carbofuran in Table 12 were 1.7 × 106 and 3.5 × 105 M−1 min−1,
respectively, for eel AChE, with carbofuran being more inhibitory than
3-OH carbofuran. The Kd and k2 values for the inhibition of bovine eryth-
rocyte AChE by carbofuran at concentrations of 2.5 × 10−6, 2.5 × 10−5, and
2.5 × 10−4 M were 3.5 × 10−6, 3.9 × 10−6, and 4.0 × 10−6 M, and 1.63, 1.55, and
1.32 min−1, respectively. The corresponding ki values were 4.66 × 105,
3.97 × 105, and 3.3 × 105 M−1 min−1.
Herzsprung et al. (1992) determined the ki values for carbofuran and
3-OH carbofuran using human (see Table 13) and horse BChE. The molar
Kd values were calculated from the ki values using a k2 value of 2.0 min−1.
The Kd and k2 values for the inhibition of equine plasma BChE by carbo-
furan at concentrations of 2.5 × 10−5 and 2.5 × 10−4 M were 3.3 × 10−6 and
4.7 × 10−6 M, and 1.07 and 0.96 min−1, respectively, with ki values of 3.2 × 105
and 2.06 × 105 M−1 min−1 (Barthova et al. 1989).

Formetanate-AChE and BChE Inhibition

No ki values for AChE or BChE were found in the literature for
formetanate.

Methiocarb-AChE and BChE Inhibition

Herzsprung et al. (1992) determined the ki values for methiocarb, methio-
carb sulfoxide, and methiocarb sulfone using eel and bovine AChE. The
values shown in Table 12 are for eel AChE with methiocarb being more
inhibitory than either the sulfoxide or sulfone. Main (1979) reported a
similar value for methiocarb (see Table 12). Buronfosse et al. (1995)
reported on the inhibitory properties of methiocarb and racemic methiocarb
sulfoxide. The racemic sulfoxide was slightly less inhibitory than methiocarb
(ki = 2.16 × 105 M−1 min−1). A 10-fold difference was observed between the
ki for the (A) and (B) sulfoxide enantiomers (5.4 × 104 M−1 min−1 and
5.02 × 105 M−1 min−1, respectively). Hetnarski and O’Brien (1975a) deter-
mined Kd, k2, and ki for methiocarb using bovine erythrocyte AChE and
the zero-time method (Hart and O’Brien 1973; O’Brien 1968). The values
were 1.5 × 10−5 M, 1.93 min−1, and 1.3 × 105 M−1 min−1, respectively.
Herzsprung et al. (1992) determined the bimolecular rate constants (ki)
for methiocarb, methiocarb sulfoxide, and methiocarb sulfone using human
(see Table 13) and horse BChE. The Kd values were calculated from
ki using a value of 2.0 min−1 for k2. The Kd and k2 values for the inhibition
of equine plasma BChE by methiocarb at concentrations of 2.5 × 10−4
and 2.5 × 10−3 M were 1.0 × 10−3 and 8.2 × 10−4 M and 1.21 and 1.13 min−1,
respectively (Barthova et al. 1989). The ki values were 1.21 × 105 and
1.38 × 105 M−1 min−1, respectively.
118 J.B. Knaak et al.

Methomyl-AChE and BChE Inhibition

Table 12 gives the ki values for methomyl using AChE (Dawson 1995;
Herzsprung et al. 1992; Vilarem et al. 1989). The Z isomer was more inhibi-
tory than racemic methomyl (Vilarem et al. 1989). Methomyl was less
inhibitory against human BChE than eel AChE as indicated in Table 13
(Herzsprung et al. 1992).

Oxamyl-AChE and BChE Inhibition

The ki for oxamyl is similar to that of the Z isomer of methomyl (i.e.,
1.31 × 105 M−1 min−1 vs. 1.14 × 105 M−1 min−1, respectively), as shown in Table
12 (Vilarem et al. 1989). Oxamyl is more inhibitory toward human BChE
than methomyl, as shown in Table 13 (Herzsprung et al. 1992).

Pirimicarb-AChE and BChE Inhibition

No ki values for AChE or BChE were found in the literature for
pirimicarb.

Propoxur-AChE and BChE Inhibition

Hetnarski and O’Brien (1975a) determined Kd, k2, and ki for propoxur using
bovine erythrocyte AChE and the zero-time method (Hart and O’Brien
1973; O’Brien 1968). The values were 2.0 × 10−5 M, 1.47 min−1, and
7.35 × 104 M−1min−1, respectively (see Table 12). According to Herzsprung
et al. (1992), propoxur is considerably more inhibitory than 3-OH propoxur
to eel AChE (Table 12). Propoxur and 2-OH propoxur are less inhibitory
to human BChE as shown by their ki values (see Table 13).

Thiodicarb-AChE and BChE Inhibition

No ki values for AChE or BChE were found in the literature for thiodicarb.
However, thiodicarb is metabolized to methomyl in the liver, and the ki
value for this carbamate may be used in PBPK/PD models.

VII. Discussion
Metcalf and March (1950), Gysin (1954), and Gubler et al. (1968) were
largely responsible for starting the development of carbamates as insect
control agents in the U.S. and Europe. Gysin (1954) concentrated on the
development of dimethylcarbamates (Isolan, Dimetan, and Dimetilan)
while Metcalf and March (1950) concentrated on the less stable but more
active methylcarbamates. The research activities in the laboratories of
Metcalf, March and Gysin, and others were ultimately responsible for the
development, registration, and marketing of the 10 carbamate insecticides
(i.e., aldicarb, carbaryl, carbofuran, formetanate, methiocarb, methomyl,
 Parameters for Carbamate Models 119

oxamyl, pirimicarb, propoxur and thiodicarb) by chemical corporations that

established agricultural chemical companies such as Union Carbide, Dow,
Dupont, Bayer, and others. The WHO’s alternative insecticide program
supported the work of Metcalf at the University of Illinois and that of
Metcalf and March at the University of California (UC) at Riverside along
with a large number of cooperating laboratories in the U.S. and throughout
the world (Wright 1971).
Oxime carbamates were developed by Union Carbide (aldicarb) and
Dupont (methomyl and oxamyl). The structural rigidity of the C=N bond
led to the formation of pairs of syn- and anti isomers in which the oximino
oxygen is either cis or trans to the aldehyde H. The syn or Z isomer is more
toxic to the housefly.
fl The toxicity of these two isomers has not been con-
sidered in environmental studies. QSAR equations were developed by
Metcalf and Fukuto at UC Riverside to assist them in understanding the
relationship between the structure of the various carbamates and their
insecticidal properties, ChE inhibition.
Goldblum et al. (1981) used the flyfl head I50 ChE data from UC Riverside
to construct new models. The synthesis and testing of new carbamates
encouraged others to examine the relationship between structure and ChE
inhibition. Hastings et al. (1970), Main and Dauterman (1963), O’Brien
(1960), and O’Brien (1976), and others showed that carbamates may be
considered strong inhibitors of AChE, BChE, and CaE or substrates with
very slow turnover rates. The dissociation rate of the inhibitor–enzyme
complex is increased when it is diluted with saline, making it difficult
fi to
measure the initial amount of inhibited enzyme. Methods involving long
incubation periods (i.e., Michel or Δ pH method; O’Brien 1960) also allow
the inhibited enzyme to recover resulting in poor estimates of inhibition.
On the basis of blood ChE measurements made according to Williams
et al. (1957) during a 2-yr carbaryl feeding study, carbaryl was considered
to be a poor ChE inhibitor (Carpenter et al. 1961). In another study in the
same laboratory, aldicarb antiChE activity was analyzed using an automatic
recording titrator (pH stat method; Knaak et al. 1966). ACh bromide was
used as a substrate immediately after a pH adjustment was made in the
reaction vessel by the titrator. Washing red cells with saline before analysis
resulted in less inhibition and was avoided (Knaak et al. 1966).
A poor understanding of the chronic toxicity of aldicarb resulted in
lowering the dose levels several times in a 2-yr feeding study, even with
AChE inhibition data being available from 30- and 90-d studies. A NOAEL
of 0.125 mg kg−1 bw was reported in a 6-mon feeding study (USEPA 1988).
A reference dose (RfD) of 0.0013 mg kg−1 bw was established by EPA using
an uncertainty factor of 100. A well-constructed PBPK/PD model would
have been useful in setting up dose levels for a 2-yr study and predicting a
NOAEL. A QSAR model, using TOPKAT, provides information on the
oral, but not dermal, toxicity of the carbamate pesticides. A considerable
amount of the in vitro work necessary for modeling carbamates [i.e., ki,
120 J.B. Knaak et al.

specific
fi content of AChE, BChE, and CaE in tissues, skin permeation con-
stants (Kp), PCt:b, Vmax, and Km] including model development, e.g., with
ACSL and ERDEM, was carried out after the carbamates were registered
for use.
The pesticide registration requirements (Knaak et al. 1993) enforced by
EPA via FIFRA require the development of considerable toxicological and
metabolic data for assessing risks to the environment and to human and
animal health. Human health risk assessments are the most diffi ficult, involv-
ing the integration of toxicological, pathological, and metabolic data into a
well-connected seamless story. Pathological data are easily integrated into
toxicological reports involving the effects of chronic exposure to pesticides,
but no one knows exactly what to do with ADME. Consequently, the
development of metabolic data is still considered to be supplementary and
not very useful until it is placed in the framework of a predictive PBPK/PD
model. FIFRA does not require the development of the parameters (i.e.,
metabolic Vmax, Km values, tissue:blood partition coefficients,
fi oral, skin, and
inhalation parameters, plasma binding measurements, etc.) necessary for
modeling insecticides. The reason for not requiring these values in the past
was based on the belief that the science of PBPK modeling was immature,
the work too costly, and not enough scientists were available to develop
data or models or both. This is certainly not the case today, based on a
monograph by Reddy et al. (2005) in which the authors presented the
results of PBPK modeling in more than 1,000 publications.
Major problems in model development involve obtaining useful Vmax and
Km values for metabolic transformations and tissue:blood partition coeffi- fi
cients for parent materials and their metabolites. Methods currently exist
for obtaining Vmax and Km from studies involving human liver microsomal
P450 and the individual CYPs (Foxenberg et al. 2006, 2007). QSAR models
for predicting metabolic rates are currently being developed by Enslein et
al. (2007). The Poulin and Theil (2002a,b) mechanistic model provides
reasonable partition coeffi ficients for nonvolatile metabolites when using log
DpH 7.4 values (ACD, Toronto, Ontario) in place of log P values.
In this review on insecticidal carbamates, a decision was made to develop
preliminary PBPK/PD models for the 10 carbamates by using published
metabolic pathway data and the multipathway PBPK/PD model of isofen-
phos (Knaak et al. 2002) as a partial template for the carbamate models.
The aldicarb PBPK/PD model is given in Fig. 9 (see Appendix C). The
conceptual models were translated into ACSL-based mathematical models
incorporating the preliminary Vmax and Km values given in Tables 24–33
(see Appendix B), and the tissue : blood partition coefficientsfi listed in
Tables 14–23 (see Appendix A) in the model. The ACSL model was com-
piled using Compaq Visual Fortran, Version 6.6A (Compaq Computer
Corporation, Houston, TX) before running the model.
The output (i.e., time tissue concentration curves, etc.) from each model
was examined to make sure the model(s) performed as desired. The meta-
 Parameters for Carbamate Models 121

bolic pathways in Tables 24–33 (Appendix B) were taken from the PBPK/
PD models and are suitable for direct entry into the front end of ERDEM
(exports command file fi to ERDEM202x) (Graphic Model). The molecular
weights for parent carbamates and their metabolites are required by
ERDEM along with PCt:b values, Vmax and Km values. The models used the
ki values for AChE and BChE in Tables 11 and 12. ki values for CaEs were
not found in the literature. ERDEM models were developed for carbofuran
(Zhang et al. 2006) and carbaryl (Okino et al. 2005a,b; Power et al. 2005)
using the metabolic pathways and preliminary Vmax and Km values pre-
sented in Tables 24–33 (Appendix B), along with the corresponding PCt:b
values and molecular weights given in Tables 14–23 (see Appendix A).
As of 2006, under the Food Quality Protection Act (FQPA) of 1996, the
EPA must consider available information concerning the cumulative effects
on human health resulting from exposure to multiple chemicals that have
a common mechanism of action, such as the N N-methyl carbamates. A cumu-
lative risk assessment also includes exposure data from multiple pathways.
EPA used the relative potency factor (RPF) method to determine the joint
risk associated with exposure to NN-methylcarbamates. Oxamyl was selected
as the index chemical for standardizing the toxic potencies and calculating
the relative potency factor for each carbamate. Brain ChE inhibition from
rats, measured at or near peak inhibition following a single oral exposure,
was used as the endpoint for measuring RPF.
In this review, preliminary PBPK/PD models were developed for the 10
N-methylcarbamates of interest. The hydrolysis of aryl N-methylcarba-
N N
mates (i.e., carbaryl, carbofuran, methiocarb, propoxur, etc.) by hydrolases
was considered to be the primary detoxification
fi pathway in humans as
opposed to the P450-catalyzed oxidative pathway leading to hydroxylated
derivatives in the rat. Hydrolytic enzymes catalyzing this pathway have not
been well characterized despite the fact that it has been more than 40 yr
since the carbamate insecticides were developed and registered. Rat and
human microsomal P450 and individual CYP studies designed to obtain
Vmax and Km data were carried out by Tang et al. (2002) and Usmani et al.
(2004a) involving carbaryl and carbofuran, respectively. Metabolic rates
were not determined for the hydrolysis of these two carbamates. The oxime
carbamates are predominantly hydrolysed to their respective oximes by
human or rat liver hydrolases. Metabolic rate constants for these enzymes
(e.g., first-order
fi or Michaelis–Menten) were not found in the literature.
Less metabolic data were found on oxamyl than on the other carbamoy-
loximes (i.e., aldicarb, methomyl, and thiodicarb). Oxamyl is metabolized
by hydrolysis to its oxime, des NN-methyl oxamyl, or by enzymatic conver-
sion via oxamyl nitrile to oxamyl acid. On the basis of the differences in
structure between the oxime and aryl carbamates, the metabolic enzymes
involved in their metabolism and their AChE ki values, it would seem
logical to use two index chemicals to represent the 10 carbamates. In
terms of oral toxicity, the oxime carbamates vary as follows: aldicarb >
122 J.B. Knaak et al.

oxamyl > methomyl > thiodicarb, with aldicarb being the most toxic and
thiodicarb the least toxic. Carbofuran is the most toxic of the aryl carba-
mates, followed by methiocarb > formetanate > propoxur > pirimicarb,
with carbaryl being the least toxic.

VIII. Recommendations
A research program is needed to develop liver microsomal metabolic rate
constants (Vmax, Km) for the hydroxylation of the aryl N-methylcarbamates
N
and the hydrolysis of the hydroxylated products and the oxime carbamates.
So far, data exist only on the formation of 3-hydroxycarbofuran from car-
bofuran and 4-hydroxycarbaryl from carbaryl. Studies involving the metab-
olism of the OP pesticides parathion and chlorpyrifos strongly suggest that
information on the activity of the individual CYPs against these insecti-
cides, and the content of these CYPs in HLMs by age, may be combined
to give the metabolic rate constants for HLMs.
A significant
fi amount of work has been carried out on the nature of the
active sites of AChE and BChE as well as the CaEs. Information regarding
the content of these enzymes in tissues needs to be improved. Current
estimates involve one study on tissue enzyme activity carried out by Maxwell
et al. (1987). A tabulation of ACh turnover numbers for AChE gives a
coefficient
fi of variation of 28.6%. The results [esterase binding site = (enzyme
activity)/turnover rate] suggest that there may be a 28% variation in the
inhibition estimates from PBPK/PD models.
Tissue:blood partition coeffi ficients estimated by mechanistic models need
to be validated based on experimental data using a high-throughput auto-
mated system based on the work of Jepson et al. (1994). To date, a system
of this nature does not exist.

Summary
A review of the scientifi fic literature was carried out for parameters [i.e.,
IC50, LD50, Kp (cm/hr) for percutaneous absorption, skin:water and tissue:
blood partition coefficients,
fi inhibition ki values, and metabolic parameters
such as Vmax and Km] suitable for building QSAR and PBPK/PD models
[using e.g., ACSL or ERDEM] for assessing human risk to 10 N-methyl-
N
carbamate insecticides: aldicarb, carbaryl, carbofuran, formetanate, methio-
carb, methomyl, oxamyl, pirimicarb, propoxur, and thiodicarb. While
searching for the parameters, PBPK/PD models were developed for these
carbamates based on published metabolic pathways. ACSL-based models
were written for each carbamate and were run using command fi files contain-
ing the parameters or parameter estimates. During this process, tissue:
blood partition coeffificients were developed for parent carbamates and
metabolites using published mechanistic models and logDpH 7.4 values. New
estimates of tissue AChE, BChE, and CaE were made using substrate
 Parameters for Carbamate Models 123

turnover numbers found in the literature. In the case of AChE, sufficient

fi
turnover data were available to calculate a coeffi
ficient of variation (28.6%).
Bimolecular inhibition rate constants (ki) involving AChE and BChE were
found for all the carbamates except pirimicarb and formetanate. Inhibition
rate constants were not found for CaEs. Several metabolic rate constants,
Vmax and Km, determined using liver microsomes and individual CYPs (P450
isozymes), were found for one or more biotransformations (i.e., hydroxyl-
ation, etc) involving carbaryl and carbofuran. Limited information is avail-
able on the enzymes carrying out hydrolysis or their rate constants. A
program is needed to develop enzyme (P450, carbamate hydrolases, etc.)
activity and enzyme content to support risk assessment methodologies
involving QSAR and PBPK/PD models.

Acknowledgments
Our thanks are extended to Prof. Herbert N. Nigg, University of Florida,
and Prof. James R. Olson, The State University of New York at Buffalo,
for reviewing the manuscript of this article and making helpful suggestions
that improve its quality. We also thank Peter Mueller and Dr. Robert W.
Gerlach, General Dynamics Information Technology, for their skilled work
in preparing this manuscript for publication. The work on this review was
funded by the U.S. EPA through General Services Administration contract
GS-09K-99-BHD-0001, tasks 9T3Z061TMA-C and EP06HO00393 with
General Dynamics Information Technology (GDIT), Henderson, NV.
Although this work was reviewed by EPA and approved for publication, it
may not necessarily reflect
fl offi
ficial Agency policy, nor does it represent the
official
fi views of GDIT. Mention of trade names or commercial products
does not constitute endorsement or recommendation for use.

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Manuscript received April 16; accepted April 21, 2007.

142 J.B. Knaak et al.

Appendix A: Chemical structures, physical parameters, and tissue:

partition coeffi
ficients of parent carbamates and metabolites.

Table 14. Chemical structure, physical and chemical properties, and tissue partition
coefficients
fi for aldicarb and the resulting metabolites.

Pesticide, pesticide ficiente

Partition coeffi
metabolite, and Physical and chemical
No.a chemical structureb propertiesc,d Tissue Rat Human

1 Aldicarb
Propanal, 2-methyl-2-(methylthio)-, O-[(methylamino) carbonyl] oxime
CAS No. 116-06-3 fat 0.97 1.00
O MW, g mol−1 190.26 brain 1.81 1.98
S
N Exp Kow @ pH 7.0 NA rapid 1.18 1.14
O N
H LogD @ pH 7.4 1.13 kidney 1.24 1.31
LogP 1.13 liver 1.19 1.54
pKa, MA 13.80 slow 1.08 1.28
WS, g L−1 (3.07) 5.31 skin 1.22 1.32
Kp, cm hr−1 NA
Log Kp NA

2 Aldicarb sulfoxide
Propanal, 2-methyl-2-(methylsulfinyl)-,
fi O-[(methylamino) carbonyl] oxime
O CAS No. 1646-87-3 fat 0.15 0.23
S
MW, g mol−1 206.26 brain 0.99 1.00
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −1.12 kidney 0.94 0.97
N
LogP −1.12 liver 0.87 0.94
O O
pKa, MA 13.57 slow 0.91 0.93
−1
HN
WS, g L (77.5) 367.54 skin 0.79 0.89

3 Aldicarb sulfone
Propanal, 2-methyl-2-(methylsulfonyl)-, O-[(methylamino) carbonyl] oxime
O CAS No. 1646-88-4 fat 0.15 0.23
S
O MW, g mol−1 222.26 brain 1.00 1.01
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −0.57 kidney 0.95 0.97
N
LogP −0.57 liver 0.87 0.95
O
pKa, MA 13.44 slow 0.91 0.94
O
WS, g L−1 (7.09) 102.70 skin 0.80 0.89
HN
 Parameters for Carbamate Models 143

Table 14. (cont.)

Pesticide, pesticide ficiente

Partition coeffi
metabolite, and Physical and chemical
No.a chemical structureb propertiesc,d Tissue Rat Human

4 Aldicarb oxime
Propanal, 2-methyl-2-(methylthio)-, oxime
OH CAS No. 1646-75-9 fat 1.15 1.18
N
MW, g mol−1 133.21 brain 1.97 2.16
Exp Kow @ pH 7.0 NA rapid 1.23 1.17
S LogD @ pH 7.4 1.21 kidney 1.30 1.37
LogP 1.21 liver 1.26 1.65
pKa, MA 11.52 slow 1.11 1.34
WS, g L−1 (12.6) 8.27 skin 1.31 1.40

5 Aldicarb oxime sulfoxide

Propanal, 2-methyl-2-(methylsulfifinyl)-, oxime
CAS No. 7635-32-7 fat 0.15 0.23
S −1
O N
MW, g mol 149.21 brain 0.99 1.00
OH Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −0.75 kidney 0.94 0.97
LogP −0.75 liver 0.86 0.94
pKa, MA 10.38 slow 0.91 0.94
WS, g L−1 (58.29) 334.23 skin 0.80 0.89

6 Aldicarb oxime sulfone

Propanal, 2-methyl-2-(methylsulfonyl)-, oxime
O CAS No. 14357-44-9 fat 0.16 0.24
S
O MW, g mol−1 165.21 brain 1.00 1.02
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
N
LogD @ pH 7.4 −0.37 kidney 0.95 0.98
OH
LogP −0.37 liver 0.87 0.96
pKa, MA 9.93 slow 0.91 0.94
WS, g L−1 L (31.86) 134.06 skin 0.80 0.90

7 Aldicarb nitrile
Propanenitrile, 2-methyl-2-(methylthio)-
S CAS No. 10074-86-9 fat 0.60 0.65
MW, g mol−1 115.19 brain 1.48 1.58
Exp Kow @ pH 7.0 NA rapid 1.08 1.08
LogD @ pH 7.4 0.90 kidney 1.12 1.17
N LogP 0.90 liver 1.06 1.30
pKa NA slow 1.00 1.14
WS, g L−1 (11.78) 17.79skin 1.05 1.15
144 J.B. Knaak et al.

Table 14. (cont.)

Pesticide, pesticide ficiente

Partition coeffi
metabolite, and Physical and chemical
No.a chemical structureb propertiesc,d Tissue Rat Human

8 Aldicarb nitrile sulfoxide

Propanenitrile, 2-methyl-2-(methylsulfifinyl)-
CAS No. 14668-28-1 fat 0.15 0.23
S −1
O
MW, g mol 131.20 brain 1.00 1.00
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −1.07 kidney 0.94 0.97
LogP −1.07 liver 0.87 0.94
pKa NA slow 0.91 0.93
WS, g L−1 (248.99) 745.74 skin 0.80 0.89

9 Aldicarb nitrile sulfone

Propanenitrile, 2-methyl-2-(methylsulfonyl)-
O CAS No. 14668-29-2 fat 0.15 0.23
N S MW, g mol−1 147.20 brain 1.00 1.00
O Exp Kow @ pH 7.0 NA rapid 0.97 0.94
LogD @ pH 7.4 −0.72 kidney 0.94 0.97
LogP −0.72 liver 0.87 0.94
pKa NA slow 0.91 0.94
WS, g L−1 (27.52) 321.51 skin 0.80 0.89

10 Aldicarb aldehyde
Propanal, 2-methyl-2-(methylthio)-
S CAS No. 16042-21-0 fat 1.10 1.13
MW, g mol−1 118.20 brain 1.93 2.12
Exp Kow @ pH 7.0 NA rapid 1.22 1.16
O LogD @ pH 7.4 1.19 kidney 1.28 1.35
LogP 1.19 liver 1.24 1.62
pKa NA slow 1.10 1.33
WS, g L−1 (12.88) 9.82 skin 1.28 1.38

11 Aldicarb aldehyde sulfoxide

2-methyl-2-(methylsulfi
finyl) propanal
CAS No. NA fat 0.15 0.23
O
S
O
MW, g mol−1 134.20 brain 1.00 1.00
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −0.78 kidney 0.94 0.97
LogP −0.78 liver 0.87 0.94
pKa NA slow 0.91 0.94
WS, g L−1 (271.30) 410.12 skin 0.80 0.89
 Parameters for Carbamate Models 145

Table 14. (cont.)

Pesticide, pesticide ficiente

Partition coeffi
metabolite, and Physical and chemical
No.a chemical structureb propertiesc,d Tissue Rat Human

12 Aldicarb aldehyde sulfone

Propanal, 2-methyl-2-(methylsulfonyl)
O CAS No. 105531-86-0 fat 0.16 0.24
S MW, g mol−1 150.20 brain 1.00 1.02
O O Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −0.40 kidney 0.95 0.98
LogP −0.40 liver 0.87 0.95
pKa NA slow 1.00 0.94
WS, g L−1 (29.16) 166.28 skin 0.80 0.90

13 Aldicarb alcohol
1-Propanol, 2-methyl-2-(methylthio)-
S CAS No. 27874-69-7 fat 0.48 0.54
MW, g mol−1 120.21 brain 1.36 1.44
Exp Kow @ pH 7.0 NA rapid 1.05 1.03
OH LogD @ pH 7.4 0.78 kidney 1.07 1.12
LogP 0.78 liver 1.01 1.21
pKa MA 14.92 slow 0.98 1.10
WS, g L−1 (61.52) 21.61 skin 0.99 1.10

14 Aldicarb alcohol sulfoxide

1-Propanol, 2-methyl-2-(methylsulfi finyl)-
CAS No. 25841-37-6 fat 0.15 0.22
S −1
O OH
MW, g mol 136.21 brain 0.98 0.99
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −1.20 kidney 0.94 0.97
LogP −1.20 liver 0.87 0.94
pKa, MA 14.02 slow 0.91 0.93
WS, g L−1 (1000.0) 919.15 skin 0.79 0.89

15 Aldicarb alcohol sulfone

1-Propanol, 2-methyl-2-(methylsulfonyl)-
O CAS No. 25841-38-7 fat 0.15 0.23
S MW, g mol−1 152.21 brain 0.99 0.99
HO O Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −0.89 kidney 0.94 0.97
LogP −0.89 liver 0.87 0.94
pKa, MA 13.67 slow 0.91 0.93
WS, g L−1 (149.79) 426.98 skin 0.79 0.89
146 J.B. Knaak et al.

Table 14. (cont.)

Pesticide, pesticide ficiente

Partition coeffi
metabolite, and Physical and chemical
No.a chemical structureb propertiesc,d Tissue Rat Human

16 Aldicarb acid
Propanoic acid, 2-methyl-2-(methylthio)-
O OH CAS No. 27868-56-0 fat 0.14 0.22
MW, g mol−1 134.20 brain 0.98 0.99
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
S LogD @ pH 7.4 −2.56 kidney 0.94 0.97
LogP 0.87 liver 0.86 0.97
pKa, MA 3.68 slow 0.91 0.93
WS, g L−1 (1000.0) 13584.03 skin 0.79 0.88

17 Aldicarb acid sulfoxide

Propanoic acid, 2-methyl-2-(methylsulfi
finyl)-
O CAS No. 3680-05-5 fat 0.14 0.22
MW, g mol−1 150.20 brain 0.98 0.99
OH NA rapid 0.94 0.94
S Exp Kow @ pH 7.0
LogD @ pH 7.4 −4.60 kidney 0.94 0.97
O
LogP −0.91 liver 0.86 0.94
pKa, MA 2.76 slow 0.91 0.93
WS, g L−1 (1000.0) 642142.60 skin 0.79 0.88

18 Aldicarb acid sulfone

Propanoic acid, 2-methyl-2-(methylsulfonyl)-
O O CAS No. 25841-43-4 fat 0.14 0.22
S MW, g mol−1 166.20 brain 0.98 0.99
O OH Exp Kow @ pH 7.0 NA rapid 0.94 0.941
LogD @ pH 7.4 −4.26 kidney 0.94 0.97
LogP −0.53 liver 0.86 0.94
pKa, MA 2.28 slow 0.91 0.93
WS, g L−1 (1000.0) 278457.10 skin 0.79 0.88
a
Parent chemical or metabolite number for cross-reference to Table 24 (Appendix B).
b
NA = Not Available. When CAS no. is available, CAS names are used. When CAS no. is
not available, IUPAC names from ACD/ChemSketch, version 8.0 (Advanced Chemistry
Development, Toronto, Ontario) are used.
c
WS (g/L) was obtained using the Syracuse Equation in Accord 6 for Excel (http://www.
accelrys.com/products/accord/accordproducts/acc_4excel.html) and Log DpH7.4 above. Values in
(. . .) after WS were calculated using ACD’s LogD Solubility Suite, version 9.0 (http://www.
acdlabs.com/products/phys_chem_lab/logd/suite.html).
d
For pKa, A = acidic, MA = more acidic, MB = more basic.
e
Tissue: blood partition coefficients.
fi
 Parameters for Carbamate Models 147

Table 15. Chemical Structure, Physical and Chemical Properties, and Tissue Parti-
tion Coeffi
ficients for Carbaryl and the Resulting Metabolites.

Pesticide, pesticide ficiente

Partition coeffi
metabolite, and Physical and
No.a chemical structureb chemical propertiesc,d Tissue Rat Human

1 Carbaryl
1-Napthalenyl methylcarbamate
CAS No. 63-25-2 fat 20.65 18.77
HN O MW, g mol−1 201.20 brain 11.14 10.07
Exp Kow @ pH 7.0 2.36 rapid 3.94 2.76
O LogD @ pH 7.4 2.40 kidney 4.56 4.06
LogP 2.40 liver 4.91 6.42
pKa, MA 12.02 slow 2.99 4.10
WS, g L−1 (0.10) 0.38 skin 6.09 4.90
Kp, cm hr−1 0.04
Log Kp −1.41

2 4-OH carbaryl
1, 4-Napthalenediol, mono (methylcarbamate)
CAS No. 5266-97-7 fat 2.62 2.55
HN O MW, g mol−1 217.22 brain 3.01 3.42
Exp Kow @ pH 7.0 1.90 rapid 1.56 1.43
O LogD @ pH 7.4 1.56 kidney 1.70 1.80
LogP 1.57 liver 1.71 2.40
pKa, MA 9.20 slow 1.24 1.78
OH
WS, g L−1 (0.32) 1.73 skin 1.90 1.96

3 5-OH carbaryl
1,5-Naphthalenediol, mono (methylcarbamate)
CAS No. 5721-72-2 fat 3.25 3.15
HN O MW, g mol−1 217.22 brain 3.55 3.88
Exp Kow @ pH 7.0 1.94 rapid 1.70 1.52
O LogD @ pH 7.4 1.66 kidney 1.86 1.95
OH LogP 1.65 liver 1.89 2.68
pKa, MA 9.02 slow 1.44 1.94
WS, g L−1 L (0.30) 1.48 skin 2.13 2.16

4 3,4-Dihydrodihydroxy carbaryl
1,2,4-Naphthalenetriol, 1,2-dihydro-, 4-(methylcarbamate)
OH CAS No. 39647-39-7 fat 1.70 1.70
HO MW, g mol−1 235.24 brain 2.42 2.67
Exp Kow @ pH 7.0 NA rapid 1.36 1.28
LogD @ pH 7.4 1.38 kidney 1.46 1.54
LogP 1.38 liver 1.44 1.96
O O pKa, MA 12.62 slow 1.20 1.52
WS, g L−1 (0.66) 1.89 skin 1.54 1.63
NH
148 J.B. Knaak et al.

Table 15. Chemical Structure, Physical and Chemical Properties, and Tissue Parti-
tion Coeffi
ficients for Carbaryl and the Resulting Metabolites.

Pesticide, pesticide ficiente

Partition coeffi
metabolite, and Physical and
No.a chemical structureb chemical propertiesc,d Tissue Rat Human

5 5,6-Dihydrodihydroxy carbaryl
1,2,5-Naphthalenetriol, 1,2-dihydro-, 5-(methylcarbamate)
CAS No. 5375-49-5 fat 0.18 0.25
HN O OH MW, g mol−1 235.24 brain 1.03 1.04
Exp Kow @ pH 7.0 NA rapid 0.95 0.95
O LogD @ pH 7.4 −0.14 kidney 0.96 0.99
OH
LogP −0.14 liver 0.89 0.97
pKa, MA 12.15 slow 0.92 0.95
WS, g L−1 (7.29) 37.54 skin 0.82 0.91

6 Hydroxymethyl carbaryl
Carbamic acid, (hydroxymethyl)-, 1-naphthalenyl ester
HO CAS No. 5266-96-6 fat 2.88 2.80
MW, g mol−1 217.22 brain 3.30 3.61
HN O Exp Kow @ pH 7.0 NA rapid 1.62 1.47
LogD @ pH 7.4 1.60 kidney 1.77 1.86
O LogP 1.60 liver 1.79 2.52
pKa, MA 10.27 slow 1.38 1.85
WS, g L−1 (0.57) 1.53 skin 2.00 2.04

7 Naphthol
1-Naphthalenol
CAS No. 90-15-3 fat 43.20 37.31
MW, g mol−1 144.17 brain 16.04 12.87
HO
Exp Kow @ pH 7.0 NA rapid 5.39 3.33
LogD @ pH 7.4 2.71 kidney 6.32 5.02
LogP 2.71 liver 6.87 8.12
pKa, MA 9.40 slow 4.01 5.08
WS, g L−1 (0.51) 0.39 skin 8.64 6.14

8 1-Hydroxy-2-naphthol
1,2-Naphthalenediol
CAS No. 547-00-5 fat 8.51 8.00
MW, g mol−1 160.17 brain 6.55 6.67
Exp Kow @ pH 7.0 NA rapid 2.58 2.08
LogD @ pH 7.4 2.04 kidney 2.93 2.91
HO LogP 2.11 liver 3.08 4.37
pKa, MA 8.42 slow 2.05 2.92
OH
WS, g L−1 L (0.95) 1.24 skin 3.69 3.40
 Parameters for Carbamate Models 149

Table 15. (cont.)

Pesticide, pesticide ficiente

Partition coeffi
metabolite, and Physical and
No.a chemical structureb chemical propertiesc,d Tissue Rat Human

9 4-OH Naphthol
1,4-Naphthalenediol
CAS No. 571-60-8 fat 5.59 5.32
MW, g mol−1 160.17 brain 5.01 5.30
HO
Exp Kow @ pH 7.0 NA rapid 2.13 1.81
LogD @ pH 7.4 1.87 kidney 2.38 2.44
OH LogP 1.88 liver 2.47 3.54
pKa, MA 10.56 slow 1.74 2.44
WS, g L−1 L (1.05) 1.73 skin 2.89 2.79

10 5-OH naphthol
1,5-Naphthalenediol
CAS No. 83-56-7 fat 6.81 6.44
MW, g mol−1 160.17 brain 5.68 5.92
HO
Exp Kow @ pH 7.0 NA rapid 2.33 1.93
LogD @ pH 7.4 1.95 kidney 2.62 2.65
OH LogP 1.96 liver 2.74 3.92
pKa, MA 9.58 slow 1.87 2.65
WS, g L−1 (0.97) 1.48 skin 3.24 3.06

11 3, 4-Dihydrodihydroxy naphthol
1, 2, 4-Naphthalenetriol, 1, 2-dihydro-
OH CAS No. 73092-92-9 fat 1.32 1.33
−1
HO MW, g mol 178.18 brain 2.11 2.33
Exp Kow @ pH 7.0 NA rapid 1.27 1.21
LogD @ pH 7.4 1.27 kidney 1.34 1.42
LogP 1.27 liver 1.31 1.75
OH pKa, MA 9.92 slow 1.14 1.40
WS, g L−1 (22.41) 4.63 skin 1.38 1.48

12 5, 6-Dihydrodihydroxy naphthol
1, 2, 5-Naphthalenetriol, 1, 2-dihydro-
OH CAS No. 5536-39-0 fat 0.64 0.69
MW, g mol−1 178.18 brain 1.51 1.63
HO Exp Kow @ pH 7.0 NA rapid 1.09 1.07
OH
LogD @ pH 7.4 0.93 kidney 1.13 1.19
LogP 0.93 liver 1.07 1.32
pKa, MA 9.72 slow 1.02 1.15
WS, g L−1 (18.72) 9.03 skin 1.07 1.17

13 Naphthyl sulfuric acid

1-Naphthalenol, hydrogen sulfate
O CAS No. 3197-94-2 fat 0.15 0.22
O S OH MW, g mol−1 224.23 brain 0.99 0.99
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O LogD @ pH 7.4 −0.97 kidney 0.94 0.97
LogP 2.53 liver 0.87 0.94
pKa, MA −4.02 slow 0.91 0.93
WS, g L−1 (331.48) 220.13 skin 0.79 0.89
150 J.B. Knaak et al.

Table 15. (cont.)

Pesticide, pesticide ficiente

Partition coeffi
metabolite, and Physical and
No.a chemical structureb chemical propertiesc,d Tissue Rat Human

14 Naphthyl glucuronic acid

β-d-Glucopyranosiduronic acid, 1-naphthalenyl
CAS No. 17238-47-0 fat 0.14 0.22
MW, g mol−1 320.29 brain 0.98 0.99
OH
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −3.31 kidney 0.94 0.97
O O
O LogP 0.40 liver 0.86 0.94
pKa, MA 2.78 slow 0.91 0.93
HO OH WS, g L−1 (1000.0) 6259.11 skin 0.79 0.88
OH

15 2-OH Naphthyl sulfuric acid

1,2-Naphthalenediol, 1-(hydrogen sulfate)
OH CAS No. 7234-05-1 fat 0.14 0.22
O
S MW, g mol−1 240.23 brain 0.98 0.99
O Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O
LogD @ pH 7.4 −2.23 kidney 0.94 0.97
LogP 1.59 liver 0.86 0.94
HO
pKa, MA −4.82 slow 0.91 0.93
WS, g L−1 (1000.0) 2148.68 skin 0.79 0.88

16 2-OH Naphthyl glucuronic acid

β-d-Glucopyranosiduronic acid, 2-hydroxy-1-naphthyl
HO O CAS No. 133102-07-5 fat 0.14 0.22
MW, g mol−1 336.29 brain 0.98 0.99
OH Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O
LogD @ pH 7.4 −4.39 kidney 0.94 0.97
LogP −0.55 liver 0.86 0.94
O OH
pKa, MA 2.74 slow 0.91 0.93
OH OH WS, g L−1 (1000.0) 42026.47 skin 0.79 0.88

17 4-OH Naphthyl sulfuric acid

1,4-Naphthalenediol, mono (hydrogen sulfate)
OH CAS No. 109841-40-9 fat 0.14 0.22
O
S MW, g mol−1 240.23 brain 0.98 0.99
O Exp Kow @ pH 7.0 NA rapid 0.94 0.94
HO O
LogD @ pH 7.4 −1.80 kidney 0.94 0.97
LogP 1.71 liver 0.86 0.94
pKa, MA −4.46 slow 0.91 0.93
WS, g L−1 (1000.0) 922.47 skin 0.79 0.88
 Parameters for Carbamate Models 151

Table 15. (cont.)

Pesticide, pesticide ficiente

Partition coeffi
metabolite, and Physical and
No.a chemical structureb chemical propertiesc,d Tissue Rat Human

18 4-OH Naphthyl glucuronic acid

3,4,5-Trihydroxy-6-[(4-hydroxy-1-naphthyl)oxy]tetrahydro-2H-pyran-2-carboxylic
H
acid
O OH CAS No. NA fat 0.14 0.22
MW, g mol−1 336.30 brain 0.98 0.99
HO Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O LogD @ pH 7.4 −4.14 kidney 0.94 0.97
LogP −0.42 liver 0.86 0.94
HO O
pKa, MA 2.76 slow 0.91 0.93
OH WS, g L−1 (1000.0) 25705.31 skin 0.79 0.88

OH

19 5-OH Naphthyl sulfuric acid

1,5-Naphthalenediol, mono (hydrogen sulfate)
CAS No. NA fat 0.14 0.22
HO
O
OH MW, g mol−1 240.23 brain 0.98 0.99
S Exp Kow @ pH 7.0 Na rapid 0.94 0.94
O
O LogD @ pH 7.4 −1.71 kidney 0.94 0.97
LogP 1.80 liver 0.86 0.94
pKa, MA −4.31 slow 0.91 0.93
WS, g L−1 (936.49) 772.84 skin 0.79 0.88

20 5-OH Naphthyl glucuronic acid

β-d-Glucopyranosiduronic acid, 5-hydroxy-1-naphthalenyl
OH CAS No. 215609-66-8 fat 0.14 0.22
HO OH MW, g mol−1 336.29 brain 0.98 0.99
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O LogD @ pH 7.4 −4.05 kidney 0.94 0.97
O O LogP −0.34 liver 0.86 0.94
OH pKa, A 2.77 slow 0.91 0.93
WS, g L−1 (1000.0) 21535.89 skin 0.79 0.88

OH

21 3,4-Dihydrodihydroxy naphthyl sulfuric acid

3,4-Dihydroxy-3,4-dihydronaphthalen-1-yl hydrogen sulfate
OH CAS No. NA fat 0.14 0.22
HO MW, g mol−1 258.25 brain 0.98 0.99
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −2.30 kidney 0.94 0.97
LogP 1.20 liver 0.86 0.94
O
O pKa, MA −3.61 slow 0.91 0.93
S WS, g L−1 (1000.0) 1959.66 skin 0.79 0.88
HO
O
152 J.B. Knaak et al.

Table 15. (cont.)

Pesticide, pesticide ficiente

Partition coeffi
metabolite, and Physical and
No.a chemical structureb chemical propertiesc,d Tissue Rat Human

22 3, 4-Dihydrodihydroxy naphthyl glucuronic acid

6-[(3,4-Dihydroxy-3,4-dihydronaphthalen-1-yl)oxy]-3,4,5-trihydroxytetrahydro-2H-
H
pyran-2-carboxyl acid
OH CAS No. NA fat 0.14 0.22
−1
HO O OH MW, g mol 354.30 brain 0.98 0.99
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O LogD @ pH 7.4 −2.94 kidney 0.94 0.97
HO OH LogP 0.78 liver 0.86 0.94
pKa, MA 2.81 slow 0.91 0.93
O OH WS, g L−1 (1000.0) 1891.33 skin 0.79 0.88

23 5, 6-Dihydrodihydroxy naphthyl sulfuric acid

5,6-Dihydroxy-5,6-dihydronaphthalen-1-yl hydrogen sulfate
CAS No. NA fat 0.14 0.22
MW, g mol−1 258.25 brain 0.98 0.99
OH
O Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −3.50 kidney 0.94 0.97
O S O
OH LogP 2.83E-4 liver 0.86 0.94
OH pKa, MA −4.10 slow 0.91 0.93
WS, g L−1 (1000.0) 20748.20 skin 0.79 0.88

24 5,6-Dihydro, 5,6-dihydroxy naphthyl glucuronic acid

6-[(5,6-Dihydroxy-5,6-dihydronaphthalen-1-yl)oxy]-3,4,5-trihydroxytetrahydro-2H-
H
pyran-2-carboxylic acid
OH CAS No. NA fat 0.14 0.22
−1
OH MW, g mol 354.31 brain 0.98 0.99
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −5.85 kidney 0.94 0.97
OH LogP −2.13 liver 0.86 0.94
HO O pKa, MA 2.78 slow 0.91 0.93
WS, g L−1 (1000.0) 577971.70 skin 0.79 0.88
O
HO

O OH

25 4-OH Carbaryl sulfuric acid

1,4-Naphthalenediol, hydrogen sulfate methylcarbamate
NH CAS No. 3197-93-1 fat 0.14 0.22
O MW, g mol−1 297.28 brain 0.98 0.99
O Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −1.93 kidney 0.94 0.97
O LogP 1.57 liver 0.86 0.94
pKa, MA −4.58 slow 0.91 0.93
O S OH
WS, g L−1 (465.72) 566.48 skin 0.79 0.88
O
 Parameters for Carbamate Models 153

Table 15. (cont.)

Pesticide, pesticide ficiente

Partition coeffi
metabolite, and Physical and
No.a chemical structureb chemical propertiesc,d Tissue Rat Human

26 4-OH Carbaryl glucuronic acid

β-d-Glucopyranosiduronic acid, 4-[[(methylamino) carbonyl] oxy]-1-naphthalenyl
OH O CAS No. 17236-46-9 fat 0.14 0.22
HO MW, g mol−1 393.34 brain 0.98 0.99
OH Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O LogD @ pH 7.4 −4.28 kidney 0.94 0.97
HO LogP −0.57 liver 0.86 0.94
O pKa, A 2.75 slow 0.91 0.93
WS, g L−1 (1000.0) 15214.21 skin 0.79 0.88

HN O

27 5-OH Carbaryl sulfuric acid

1, 5-Naphthalenediol, hydrogen sulfate methylcarbamate
NH CAS No. 24281-28-5 fat 0.14 0.22
O MW, g mol−1 297.28 brain 0.98 0.99
O Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −2.02 kidney 0.94 0.97
LogP 1.48 liver 0.86 0.94
O
pKa, MA −4.40 slow 0.91 0.93
WS, g L−1 (518.41) 676.16 skin 0.79 0.88
O S OH
O

28 5-OH Carbaryl glucuronic acid

3,4,5-Trihydroxy-6-[(5-{[(methylamino)carbonyl]oxy}-1-naphthyl)oxy]tetrahydro-2H-
H
pyran-2-carboxylic acid
O OH CAS No. NA fat 0.14 0.22
−1
OH MW, g mol 393.34 brain 0.98 0.99
HO Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O LogD @ pH 7.4 −4.37 kidney 0.94 0.97
OH LogP −0.65 liver 0.86 0.94
O pKa, A 2.77 slow 0.91 0.93
O WS, g L−1 (1000.0) 18159.73 skin 0.79 0.88

HN O
154 J.B. Knaak et al.

Table 15. (cont.)

Pesticide, pesticide ficiente

Partition coeffi
metabolite, and Physical and
No.a chemical structureb chemical propertiesc,d Tissue Rat Human

29 3, 4-Dihydrodihydroxy carbaryl sulfuric acid

3-Hydroxy-4-(sulfooxy)-3,4-dihyronaphthalen-1-yl methylcarbamate
O OH OH CAS No. NA fat 0.14 0.22
O
HN S MW, g mol−1 315.30 brain 0.98 0.99
O Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O O
LogD @ pH 7.4 −2.20 kidney 0.94 0.97
LogP 1.30 liver 0.86 0.94
pKa, MA −3.75 slow 0.91 0.93
WS, g L−1 (570.37) 755.34 skin 0.79 0.88

30 3, 4-Dihydrodihydroxy carbaryl glucuronic acid

3,4,5-Trihydroxy-6-[(2-hydroxy-4-{[(methylamino)carbonyl]oxy}-1,2-
dihydronaphthalen-1-yl)oxy]tetrahydro-2H-pyran-2-carboxylic
H acid
OH CAS No. NA fat 0.14 0.22
HO OH MW, g mol−1 411.36 brain 0.98 0.99
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O LogD @ pH 7.4 −4.70 kidney 0.94 0.97
O O LogP −0.99 liver 0.86 0.94
OH HO pKa, MA 2.81 slow 0.91 0.93
WS, g L−1 (1000.0) 26873.32 skin 0.79 0.88

O O

NH

31 5,6-Dihydrodihydroxy carbaryl sulfuric acid

6-Hydroxy-5-(sulfooxy)-5,6-dihydronaphthalen-1-yl methyl carbamate
NH CAS No. NA fat 0.14 0.22
O MW, g mol−1 315.30 brain 0.98 0.99
O Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −3.71 kidney 0.94 0.97
LogP −0.21 liver 0.86 0.94
O
pKa, MA −3.82 slow 0.91 0.93
WS, g L−1 (1000.0) 14712.28 skin 0.79 0.88
HO O S OH
O
 Parameters for Carbamate Models 155

Table 15. (cont.)

Pesticide, pesticide ficiente

Partition coeffi
metabolite, and Physical and
No.a chemical structureb chemical propertiesc,d Tissue Rat Human

32 5,6-Dihydrodihydroxy carbaryl glucuronic acid

3,4,5-Trihydroxy-6-[(2-hydroxy-5-{[(methylamino)carbonyl]oxy]oxy}-1,2-
dihydronaphthalen-1-yl)oxy]tetrahydro-2H-pyran-2-carboxylic
H acid
O OH CAS No. NA fat 0.14 0.22
−1
OH MW, g mol 411.36 brain 0.98 0.99
HO Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O LogD @ pH 7.4 −6.21 kidney 0.94 0.97
OH LogP −2.50 liver 0.86 0.94
O pKa, MA 2.80 slow 0.91 0.93
O WS, g L−1 (1000.0) 523433.20 skin 0.79 0.88

HN O OH

33 Hydroxymethyl carbaryl sulfuric acid

1-Naphthyl[(sulfooxy)methyl]carbamate
OH CAS No. NA fat 0.14 0.22
O S O MW, g mol−1 297.28 brain 0.98 0.99
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O
LogD @ pH 7.4 −1.64 kidney 0.94 0.97
LogP 1.87 liver 0.86 0.94
HN O
pKa, MA −3.90 slow 0.91 0.93
WS, g L−1 (312.32) 320.28 skin 0.79 0.88
O

34 Hydroxymethyl carbaryl glucuronic acid

3,4,5-Trihydroxy-6-({[(1-naphthyloxy)carbonyl]amino}methoxy)tetrahydro-2H-
H
pyran-2-carboxylic acid
OH O CAS No. NA fat 0.14 0.22
−1
HO MW, g mol 393.34 brain 0.98 0.99
OH Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O LogD @ pH 7.4 −4.14 kidney 0.94 0.97
HO LogP −0.42 liver 0.86 0.94
O pKa, MA 2.80 slow 0.91 0.93
WS, g L−1 (1000.0) 11554.50 skin 0.79 0.88
HN O

a
Parent chemical or metabolite number for cross-reference to Table 25 (Appendix B).
b–e
See footnotes in Table 14 (Appendix A).
156 J.B. Knaak et al.

Table 16. Chemical Structure, Physical and Chemical Properties, and Tissue Parti-
tion Coeffi
ficients for Carbofuran and the Resulting Metabolites.
e
Pesticide, pesticide Partition coefficient
fi
metabolite, and chemical Physical and chemical
No.a structureb propertiesc,d Tissue Rat Human

1 Carbofuran
7-Benzofuranol, 2,3-dihydro-2,2-dimethyl-, methylcarbamate
CAS No. 1563-66-2 fat 4.25 4.08
MW, g mol−1 221.25 brain 4.20 4.53
O Exp Kow @ pH 7.0 NA rapid 1.89 1.65
LogD @ pH 7.4 1.76 kidney 2.09 2.18
O O LogP 1.76 liver 2.15 3.08
pKa, MA 12.28 slow 1.57 2.17
NH WS, g L−1 (0.31) 1.06 skin 2.47 2.45
Kp, cm hr −1 NA
Log Kp NA

2 Carbofuran-7-phenol
7-Benzofuranol, 2,3-dihydro-2,2-dimethyl-
CAS No. 1563-38-8 fat 13.57 12.56
MW, g mol−1 164.20 brain 8.74 8.41
O Exp Kow @ pH 7.0 NA rapid 3.23 2.43
LogD @ pH 7.4 2.23 kidney 3.71 3.50
OH
LogP 2.23 liver 3.96 5.42
pKa, MA 10.09 slow 2.50 3.52
WS, g L−1 L (1.23) 0.82 skin 4.84 4.16

3 3-OH carbofuran
3,7-Benzofurandiol, 2,3-dihydro-2,2-dimethyl-, 7-(methylcarbamate)
OH CAS No. 16655-82-6 fat 0.22 0.29
MW, g mol−1 237.25 brain 1.08 1.11
Exp Kow @ pH 7.0 NA rapid 0.97 0.96
O
LogD @ pH 7.4 0.21 kidney 0.98 1.01
LogP 0.21 liver 0.90 1.01
O O pKa, MA 12.28 slow 0.93 0.98
WS, g L−1 (3.87) 18.38 skin 0.84 0.94
NH

4 Hydroxymethyl carbofuran
Carbamic acid, (hydroxymethyl)-, 2,3-dihydro-2,2-dimethyl-7-benzofuranyl ester
CAS No. 18999-70-7 fat 0.68 0.73
MW, g mol−1 237.25 brain 1.55 1.68
O Exp Kow @ pH 7.0 NA rapid 1.11 1.08
LogD @ pH 7.4 0.96 kidney 1.14 1.20
O O LogP 0.96 liver 1.09 1.35
pKa, MA 10.53 slow 1.02 1.17
NH WS, g L−1 (1.83) 4.21 skin 1.09 1.19

OH
 Parameters for Carbamate Models 157

Table 16. (cont.)

e
Pesticide, pesticide Partition coefficient
fi
metabolite, and chemical Physical and chemical
No.a structureb propertiesc,d Tissue Rat Human

5 Carbofuran-3,7-diol
3,7-Dihydroxy-2,2-dimethyl-2,3-dihydrobenzofuran
OH CAS No. 17781-15-6 fat 0.40 0.47
MW, g mol−1 180.20 brain 1.28 1.35
Exp Kow @ pH 7.0 NA rapid 1.02 1.01
O
LogD @ pH 7.4 0.68 kidney 1.05 1.09
LogP 0.68 liver 0.98 1.16
OH pKa, MA 9.90 slow 0.97 1.06
WS, g L−1 (15.74) 14.43 skin 0.94 1.04

6 3-Keto carbofuran
3(2H)-Benzofuranone,
H 2,2-dimethyl-7-[[(methylamino) carbonyl] oxy]-
O CAS No. 16709-30-1 fat 1.96 1.94
MW, g mol−1 235.24 brain 2.62 2.90
Exp Kow @ pH 7.0 NA rapid 1.42 1.32
O
LogD @ pH 7.4 1.44 kidney 1.53 1.62
LogP 1.44 liver 1.52 2.09
O O pKa, MA 11.92 slow 1.24 1.60
WS, g L−1 (0.19) 1.68 skin 1.65 1.73
NH

7 3-Keto-7-phenol
7-Hydroxy-2,2-dimethylbenzofuran-3-one
O CAS No. 17781-16-7 fat 7.90 7.44
MW, g mol−1 178.18 brain 6.25 6.41
Exp Kow @ pH 7.0 NA rapid 2.49 2.03
O
LogD @ pH 7.4 2.01 kidney 2.82 2.82
LogP 2.05 liver 2.96 4.22
OH pKa, MA 8.44 slow 1.99 2.83
WS, g L−1 (0.73) 1.08 skin 3.54 3.28

8 7-Phenyl sulfuric acid

7-Benzofuranol, 2,3-dihydro-2,2-dimethyl-, hydrogen sulfate
CAS No. 70988-92-0 fat 0.14 0.22
MW, g mol−1 244.27 brain 0.98 0.99
O Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O LogD @ pH 7.4 −1.59 kidney 0.94 0.97
O
S LogP 1.91 liver 0.86 0.94
HO pKa, MA −3.96 slow 0.91 0.93
O
WS, g L−1 (1000.0) 579.80 skin 0.79 0.88
158 J.B. Knaak et al.

Table 16. (cont.)

e
Pesticide, pesticide Partition coefficient
fi
metabolite, and chemical Physical and chemical
No.a structureb propertiesc,d Tissue Rat Human

9 7-Phenyl glucuronic acid

β-d-Glucopyranosiduronic acid, 2,3-dihydro-2,2-dimethyl -7-benzofuranyl
CAS No. 70988-91-9 fat 0.14 0.22
MW, g mol−1 340.33 brain 0.98 0.99
OH O Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −3.94 kidney 0.94 0.97
O O LogP −0.23 liver 0.86 0.94
O
pKa, MA 2.79 slow 0.91 0.93
HO OH WS, g L−1 (1000.0) 16414.89 skin 0.79 0.88

OH

10 3-OH carbofuran sulfuric acid

3,7-Benzofurandiol, 2,3-dihydro-2,2-dimethyl-, 3-(hydrogen sulfate)
7-(methylcarbamate)
O CAS No. 70988-90-8 fat 0.14 0.22
HN MW, g mol−1 317.32 brain 0.98 0.99
O Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O OH LogD @ pH 7.4 −3.00 kidney 0.94 0.97
S LogP 0.50 liver 0.86 0.94
O pKa, MA −4.03 slow 0.91 0.93
O O
WS, g L−1 (1000.0) 3544.26 skin 0.80 0.88

11 3-OH carbofuran glucuronic acid

β-d-Glucopyranosiduronic acid, 2,3-dihydro-2,2-dimethyl-7-[[(methylamino) carbonyl
] oxy]-3-benzofuranyl
O CAS No. 53305-32-1 fat 0.15 0.22
OH MW, g mol−1 413.38 brain 0.98 0.99
HO
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O LogD @ pH 7.4 −5.50 kidney 0.94 0.97
HO LogP −1.79 liver 0.86 0.94
O
pKa, A 2.79 slow 0.91 0.93
HO WS, g L−1 (1000.0) 125839.90 skin 0.79 0.88

O O

NH
 Parameters for Carbamate Models 159

Table 16. (cont.)

e
Pesticide, pesticide Partition coefficient
fi
metabolite, and chemical Physical and chemical
No.a structureb propertiesc,d Tissue Rat Human

12 Hydroxymethyl carbofuran sulfate

2,2-Dimethyl-2,3-dihydro-1-benzofuran-7-yl [(sulfooxy) methyl] carbamate
HO CAS No. NA fat 0.14 0.22
O
S MW, g mol−1 317.32 brain 0.98 0.99
O Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O O
HN
LogD @ pH 7.4 −2.27 kidney 0.94 0.97
LogP 1.23 liver 0.86 0.94
O
pKa, A −3.87 slow 0.91 0.93
WS, g L−1 (1000.0) 843.54 skin 0.79 0.88
O

13 Hydroxymethyl carbofuran glucuronic acid

({[(2,2-dimethylhexahydro-2H-cyclopenta[b]furan-6-yl)oxy]carbonyl}amino)
H
methyl hexopyranosiduronic acid
CAS No. NA fat 0.14 0.22
MW, g mol−1 413.38 brain 0.98 0.99
O Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −4.77 kidney 0.94 0.97
O O
LogP −1.06 liver 0.86 0.94
pKa, MA 2.80 slow 0.91 0.93
HN
OH WS, g L−1 (1000.0) 29959.0 skin 0.79 0.88
O OH

O
OH

HO O

14 3,7-Diol sulfuric acid

3,7-Benzofurandiol, 2,3-dihydro-2,2-dimethyl-, 7-(hydrogen sulfate)
OH CAS No. 90433-38-8 fat 0.14 0.22
MW, g mol−1 260.26 brain 0.98 0.99
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O
LogD @ pH 7.4 −3.14 kidney 0.94 0.97
LogP 0.36 liver 0.86 0.94
O
O pKa, MA −3.96 slow 0.91 0.93
S WS, g L−1 (1000.0) 9955.20 skin 0.80 0.88
HO
O
160 J.B. Knaak et al.

Table 16. (cont.)

e
Pesticide, pesticide Partition coefficient
fi
metabolite, and chemical Physical and chemical
No.a structureb propertiesc,d Tissue Rat Human

15 3,7-Diol glucuronic acid

β-d-Glucopyranosiduronic acid, 2,3-dihydro-3-hydroxy-2,2-dimethyl-7-benzofuranyl
OH CAS No. 90433-37-7 fat 0.14 0.22
MW, g mol−1 356.32 brain 0.98 0.99
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O
LogD @ pH 7.4 −5.49 kidney 0.94 0.97
OH
LogP −1.78 liver 0.86 0.94
O O pKa, MA 2.79 slow 0.91 0.93
O
WS, g L−1 (1000.0) 276956.70 skin 0.79 0.88

HO OH
OH

16 3-Keto-7-phenyl sulfuric acid

3(2H)-Benzofuranone,
H 2,2-dimethyl-7-(sulfooxy)-
O CAS No. 70988-94-2 fat 0.14 0.22
MW, g mol−1 258.25 brain 0.98 0.99
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O
LogD @ pH 7.4 −1.98 kidney 0.94 0.97
LogP 1.52 liver 0.86 0.94
O
O pKa, MA −4.34 slow 0.91 0.93
S WS, g L−1 (1000.0) 1045.15 skin 0.80 0.88
HO
O

17 3-Keto-7-phenol glucuronic acid

β-d-Glucopyranosiduronic acid, 2,3-dihydro-2,2-dimethyl-3-oxo-7-benzofuranyl
O CAS No. 70988-93-1 fat 0.14 0.22
MW, g mol−1 354.31 brain 0.98 0.99
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O
LogD @ pH 7.4 −4.33 kidney 0.94 0.97
OH LogP −0.61 liver 0.86 0.94
O O pKa, MA 2.77 slow 0.91 0.93
O
WS, g L−1 (1000.0) 29099.61 skin 0.79 0.88

HO OH
OH

a
Parent chemical or metabolite number for cross-reference to Table 26 (Appendix B).
b–e
See footnotes in Table 14 (Appendix A).
Table 17. Chemical Structure, Physical and Chemical Properties, and Tissue Parti-
tion Coeffi
ficients for Formetanate and the Resulting Metabolites.
e
Pesticide, pesticide Partition coefficient
fi
metabolite, and chemical Physical and chemical
No.a structureb propertiesc,d Tissue Rat Human

1 Formetanate
Methanimidamide, N,N-dimethyl-
N N -[3-[[(methylamino) carbonyl] oxy]phenyl]-
N′
CAS No. 22259-30-9 fat 0.48 0.54
N MW, g mol−1 221.26 brain 1.36 1.44
Exp Kow @ pH 7.0 NA rapid 1.05 1.03
N LogD @ pH 7.4 0.78 kidney 1.07 1.12
LogP 0.86 liver 1.01 1.21
pKa, MA 12.36 slow 0.98 1.09
WS, g L−1 (1.55) 7.31 skin 0.99 1.09
O O Kp, cm hr−1 NA
Log Kp NA
HN

2 Demethylformetanate
Methanimidamide, N-methyl-
N N -[3-[[(methylamino) carbonyl] oxy] phenyl]-
N′
CAS No. 25636-15-1 fat 0.17 0.24
HN MW, g mol−1 207.23 brain 1.01 1.03
Exp Kow @ pH 7.0 NA rapid 0.95 0.95
N LogD @ pH 7.4 −0.29 kidney 0.95 0.98
LogP 0.58 liver 0.88 0.96
pKa, MA 12.38 slow 0.91 0.95
WS, g L−1 (35.16) 71.03 skin 0.81 0.90
O O

HN

3 3-Formaminophenyl N-methylcarbamate
Formamide, N
N-[3-[[(methylamino) carbonyl] oxy] phenyl]-
O CAS No. 24891-34-7 fat 0.28 0.35
MW, g mol−1 194.19 brain 1.15 1.20
HN Exp Kow @ pH 7.0 NA rapid 0.99 0.98
LogD @ pH 7.4 0.44 kidney 1.00 1.04
LogP 0.44 liver 0.93 1.06
pKa, MA 12.11 slow 0.94 1.01
WS, g L−1 (3.65) 19.71 skin 0.88 0.98
O O

HN

4 3-Formaminophenol
Formamide, N-(3-hydroxyphenyl)-
N
O CAS No. 24891-35-8 fat 0.49 0.55
MW, g mol−1 137.14 brain 1.37 1.45
HN Exp Kow @ pH 7.0 NA rapid 1.05 1.03
LogD @ pH 7.4 0.79 kidney 1.08 1.13
LogP 0.80 liver 1.02 1.22
pKa, MA 9.41 slow 0.99 1.10
OH WS, g L−1 (5.68) 18.20 skin 0.99 1.09

161
162 J.B. Knaak et al.

Table 17. (cont.)

e
Pesticide, pesticide Partition coefficient
fi
metabolite, and chemical Physical and chemical
No.a structureb propertiesc,d Tissue Rat Human

5 3-Aminophenol
Phenol, 3-amino-
H2N CAS No. 591-27-5 fat 0.25 0.32
MW, g mol−1 109.13 brain 1.12 1.15
Exp Kow @ pH 7.0 NA rapid 0.98 0.97
LogD @ pH 7.4 0.34 kidney 0.99 1.02
OH LogP 0.34 liver 0.92 1.04
pKa, MA 10.01 slow 0.93 0.99
WS, g L−1 (21.55) 56.12 skin 0.86 0.96

6 3-Acetamidophenol
Acetamide, N-(3-hydroxyphenyl)-
N
O CAS No. 621-42-1 fat 0.44 0.50
MW, g mol−1 151.16 brain 1.32 1.40
HN Exp Kow @ pH 7.0 NA rapid 1.04 1.02
LogD @ pH 7.4 0.73 kidney 1.06 1.11
LogP 0.73 liver 1.00 1.18
pKa, MA 9.50 slow 0.98 1.07
OH WS, g L−1 (12.92) 17.84 skin 0.97 1.06

7 3-Formaminophenyl glucuronic acid

6-[3-(Formylamino)phenoxy]-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic
H acid
H CAS No. NA fat 0.14 0.22
O N MW, g mol−1 313.26 brain 0.98 0.99
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −5.27 kidney 0.94 0.97
OH
LogP −1.56 liver 0.86 0.94
O OH pKa, MA 2.78 slow 0.91 0.93
WS, g L−1 (1000.0) 325042.00 skin 0.79 0.88
O
OH

HO O

8 3-Formaminophenyl sulfuric acid

3-(Formylamino)phenyl hydrogen sulfate
O CAS No. NA fat 0.14 0.22
MW, g mol−1 217.20 brain 0.98 0.99
HO S O
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O LogD @ pH 7.4
O −2.92 kidney 0.94 0.97
NH LogP 0.58 liver 0.86 0.94
pKa, MA −4.14 slow 0.91 0.93
WS, g L−1 (1000.0) 11100.29 skin 0.79 0.88
 Parameters for Carbamate Models 163

Table 17. (cont.)

e
Pesticide, pesticide Partition coefficient
fi
metabolite, and chemical Physical and chemical
No.a structureb propertiesc,d Tissue Rat Human

9 3-Aminophenyl glucuronic acid

β-d-Glucopyranosiduronic acid, m-aminophenyl
H2N CAS No. 108595-12-6 fat 0.14 0.22
MW, g mol−1 285.25 brain 0.98 0.99
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
OH LogD @ pH 7.4 −5.84 kidney 0.94 0.97
O OH LogP −2.13 liver 0.86 0.94
pKa, MA 2.80 slow 0.91 0.93
O
WS, g L−1 (1000.0) 1451978.00 skin 0.79 0.88
OH

HO O

10 3-Aminophenyl sulfuric aicd

Phenol, 3-amino-, hydrogen sulfate
O CAS No. 27991-69-1 fat 0.14 0.22
MW, g mol−1 189.19 brain 0.98 0.99
HO S O
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O LogD @ pH 7.4 −3.5 kidney 0.94 0.97
NH2 LogP −0.003 liver 0.86 0.94
pKa, MA −3.78 slow 0.91 0.93
WS, g L−1 (1000.0) 48373.21 skin 0.79 0.88

11 3-Acetamidophenyl glucuronic acid

β-d-Glucopyranosiduronic acid, 3-(acetylamino) phenyl
H CAS No. 102989-23-1 fat 0.14 0.22
N MW, g mol−1 327.79 brain 0.98 0.99
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O LogD @ pH 7.4 −5.48 kidney 0.94 0.97
OH
LogP −1.62 liver 0.86 0.94
O OH pKa, MA 2.78 slow 0.91 0.93
WS, g L−1 (1000.0) 308058.70 skin 0.79 0.88
O
OH

HO O

12 3-Acetamidophenyl sulfuric acid

Acetamide, N
N-[3-(sulfooxy) phenyl]-
O CAS No. 52175-03-8 fat 0.14 0.22
MW, g mol−1 231.23 brain 0.98 0.99
HO S O O
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O LogD @ pH 7.4 −2.99 kidney 0.94 0.97
NH LogP 0.51 liver 0.86 0.94
pKa, MA −4.11 slow 0.91 0.93
WS, g L−1 (1000.0) 10719.42 skin 0.79 0.88

a
Parent chemical or metabolite number for cross-reference to Table 27 (Appendix B).
b–e
See footnotes in Table 14 (Appendix A).
164 J.B. Knaak et al.

Table 18. Chemical Structure, Physical and Chemical Properties, and Tissue Parti-
tion Coeffi
ficients for Methiocarb and the Resulting Metabolites.
e
Pesticide, pesticide Partition coefficient
fi
metabolite, and chemical Physical and chemical
No.a structureb propertiesc,d Tissue Rat Human

1 Methiocarb
Phenol, 3,5-dimethyl-4-(methylthio)-, methylcarbamate
CAS No. 2032-65-7 fat 64.84 53.51
HN O MW, g mol−1 225.31 brain 18.85 14.20
Exp Kow @ pH 7.0 NA rapid 6.22 3.60
O LogD @ pH 7.4 2.89 kidney 7.32 5.47
LogP 2.89 liver 7.99 8.92
pKa, MA 12.16 slow 4.58 5.55
S WS, g L−1 (0.12) 0.11 skin 10.11 6.72
Kp, cm hr−1 NA
Log Kp NA

2 Methiocarb sulfoxide
Phenol, 3,5-dimethyl-4-(methylsulfinyl)-,
fi methylcarbamate
CAS No. 2635-10-1 fat 0.37 0.43
HN O MW, g mol−1 241.31 brain 1.24 1.31
Exp Kow @ pH 7.0 NA rapid 1.01 1.00
O LogD @ pH 7.4 0.62 kidney 1.03 1.08
LogP 0.62 liver 0.97 1.13
O pKa, MA 12.03 slow 0.96 1.04
S WS, g L−1 (1.79) 7.81 skin 0.93 1.02

3 Methiocarb sulfone
Phenol, 3,5-dimethyl-4-(methylsulfonyl)-, methylcarbamate
CAS No. 2179-25-1 fat 1.29 1.31
HN O MW, g mol−1 257.31 brain 2.09 2.30
Exp Kow @ pH 7.0 NA rapid 1.26 1.20
O LogD @ pH 7.4 1.26 kidney 1.34 1.41
LogP 1.26 liver 1.31 1.73
O pKa, MA 11.96 slow 1.13 1.39
S WS, g L−1 (0.46) 1.81 skin 1.37 1.46
O

4 Hydroxymethyl methiocarb
3,5-Dimethyl-4-(methylthio) phenyl hydroxymethyl) carbamate
HO CAS No. NA fat 9.63 9.02
MW, g mol−1 241.31 brain 7.08 7.11
HN O Exp Kow @ pH 7.0 NA rapid 2.74 2.17
LogD @ pH 7.4 2.09 kidney 3.12 3.06
O LogP 2.09 liver 3.30 4.64
pKa, MA 10.42 slow 2.16 3.07
WS, g L−1 (0.74) 0.43 skin 3.97 3.59
S
 Parameters for Carbamate Models 165

Table 18. (cont.)

e
Pesticide, pesticide Partition coefficient
fi
metabolite, and chemical Physical and chemical
No.a structureb propertiesc,d Tissue Rat Human

5 Methiocarb phenol
Phenol, 3,5-dimethyl-4-(methylthio)-
HO CAS No. 7379-51-3 fat 45.24 38.89
MW, g mol−1 168.26 brain 16.36 13.03
Exp Kow @ pH 7.0 NA rapid 5.48 3.36
S LogD @ pH 7.4 2.73 kidney 6.43 5.07
LogP 2.73 liver 6.99 8.22
pKa, MA 10.03 slow 4.07 5.14
WS, g L−1 (0.84) 0.29 skin 8.81 6.21

6 Methiocarb phenol sulfoxide

Phenol, 3,5-dimethyl-4-(methylsulfinyl)-
fi
HO CAS No. 22454-92-8 fat 1.15 1.18
MW, g mol−1 184.26 brain 1.97 2.16
O Exp Kow @ pH 7.0 NA rapid 1.23 1.17
S LogD @ pH 7.4 1.21 kidney 1.29 1.37
LogP 1.23 liver 1.26 1.65
pKa, MA 8.64 slow 1.11 1.34
WS, g L−1 (6.87) 4.86 skin 1.31 1.40

7 Methiocarb phenol sulfone

Phenol, 3,5-dimethyl-4-(methylsulfonyl)-;
3,5-Xylenol, 4-(methylsulfonyl)-;
OH CAS No. 14763-62-3 fat 0.75 0.79
MW, g mol−1 200.26 brain 1.62 1.75
Exp Kow @ pH 7.0 NA rapid 1.12 1.09
LogD @ pH 7.4 1.01 kidney 1.17 1.23
O LogP 1.15 liver 1.12 1.40
S pKa, MA 7.81 slow 1.04 1.20
O
WS, g L−1 (4.08) 5.98 skin 1.12 1.22

8 Hydroxymethyl methiocarb glucuronic acid

({[3,5-Dimethyl-4-(methylthio)phenoxy]carbonyl}amino)methyl
hexopyranosiduronic acid
OH H CAS No. NA fat 0.14 0.22
O O N O MW, g mol−1 417.43 brain 0.98 0.99
O
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O
HO OH S LogD @ pH 7.4 −3.65 kidney 0.94 0.97
OH LogP 0.07 liver 0.86 0.94
pKa, MA 2.8 slow 0.91 0.93
WS, g L−1 (1000.0) 3124.6 skin 0.79 0.85
166 J.B. Knaak et al.

Table 18. (cont.)

e
Pesticide, pesticide Partition coefficient
fi
metabolite, and chemical Physical and chemical
No.a structureb propertiesc,d Tissue Rat Human

9 Hydroxymethyl methiocarb sulfuric acid

3,5-Dimethyl-4-(methylthio) phenyl [(sulfooxy) methyl] carbamate
O CAS No. NA fat 0.14 0.22
−1
O S OH MW, g mol 321.37 brain 0.99 0.99
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
HN O LogD @ pH 7.4 −1.15 kidney 0.94 0.97
O
LogP 2.36 liver 0.86 0.94
O pKa, MA −3.88 slow 0.91 0.93
WS, g L−1 (177.18) 88.21 skin 0.79 0.89

10 Methiocarb phenyl glucuronic acid

6-[3,5-Dimethyl-4-(methylthio) phenoxy]-3,4,5-trihydroxytetrahydro-2H-pyran-2-
H
carboxylic acid
OH O CAS No. NA fat 0.14 0.22
MW, g mol−1 344.39 brain 0.98 0.99
HO
OH Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −3.11 kidney 0.94 0.97
O
HO LogP 0.60 liver 0.86 0.94
pKa, MA 2.78 slow 0.91 0.93
O WS, g L−1 (1000.0) 3033.05 skin 0.79 0.88

11 Methiocarb phenyl sulfuric acid

3,5-Dimethyl-4-(methylthio) phenyl hydrogen sulfate
OH CAS No. NA fat 0.14 0.22
MW, g mol−1 248.32 brain 0.99 1.00
O S O
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O LogD @ pH 7.4 −0.76 kidney 0.94 0.97
LogP 2.74 liver 0.87 0.94
pKa, MA −4.18 slow 0.91 0.94
S WS, g L−1 (309.93) 107.72 skin 0.79 0.88
 Parameters for Carbamate Models 167
Table 18. (cont.)
e
Pesticide, pesticide Partition coefficient
fi
metabolite, and chemical Physical and chemical
No.a structureb propertiesc,d Tissue Rat Human

12 Methiocarb sulfoxide phenyl glucuronic acid

6-[3,5-Dimethyl-4-(methylsulfi
finyl)phenoxy]-3,4,5-trihyroxytetrahydro-2H-pyran-2-
H
carboxylic acid
OH O CAS No. NA fat 0.14 0.22
MW, g mol−1 360.38 brain 0.98 1.00
HO
OH Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −5.09 kidney 0.94 0.97
O
HO LogP −1.38 liver 0.86 0.94
pKa, MA 2.78 slow 0.91 0.93
O WS, g L−1 (1000.0) 119139.10 skin 0.79 0.88

O
S

13 Methiocarb sulfoxide phenyl sulfuric acid

3,5-Dimethyl-4-(methylsulfinyl)phenyl
fi hydrogen sulfate
OH CAS No. NA fat 0.14 0.22
MW, g mol−1 264.32 brain 0.98 1.00
O S O
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O LogD @ pH 7.4 −2.75 kidney 0.94 0.97
LogP 0.75 liver 0.86 0.94
O pKa, MA −4.39 slow 0.91 0.93
S WS, g L−1 (1000.0) 4389.26 skin 0.79 0.88

14 Methiocarb sulfone phenyl glucuronic acid

6-[3,5-Dimethyl-4-(methylsulfonyl) phenoxy]-3,4,5-trihydroxytetrahydro-2H-pyran-2-
H
carboxylic acid
HO OH CAS No. NA fat 0.14 0.22
MW, g mol−1 376.38 brain 0.98 1.00
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O OH
LogD @ pH 7.4 −4.74 kidney 0.94 0.97
O
LogP −1.03 liver 0.86 0.94
O pKa, MA 2.77 slow 0.91 0.93
HO WS, g L−1 (1000.0) 47813.03 skin 0.79 0.88
O
S
O

15 Methiocarb sulfone phenyl sulfuric acid

3,5-Dimethyl-4-(methylsulfonyl)phenyl hydrogen sulfate
OH CAS No. NA fat 0.14 0.22
MW, g mol−1 280.32 brain 0.98 0.99
O S O
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O LogD @ pH 7.4 −2.39 kidney 0.94 0.97
LogP 1.11 liver 0.86 0.94
O
pKa, MA −4.51 slow 0.91 0.93
S WS, g L−1 (1000.0) 1753.91 skin 0.79 0.88
O

a
Parent chemical or metabolite number for cross-reference to Table 28 (Appendix B).
b–e
See footnotes in Table 14 (Appendix A).
168 J.B. Knaak et al.

Table 19. Chemical Structure, Physical and Chemical Properties, and Tissue Parti-
tion Coeffi
ficients for Methomyl and the Resulting Metabolites.

Pesticide, pesticide ficiente

Partition coeffi
metabolite, and Physical and chemical
No.a chemical structureb propertiesc,d Tissue Rat Human

1 syn-(Z)-methomyl
Ethanimidothioic acid, N
N-[[(methylamino)carbonyl]oxy]-methyl ester, (Z)-
O CAS No. 19928-37-1 fat 0.36 0.42
MW, g mol−1 162.21 brain 1.23 1.29
O N Exp Kow @ pH 7.0 NA rapid 1.01 1.00
H
S N LogD @ pH 7.4 0.60 kidney 1.03 1.07
LogP 0.60 liver 0.96 1.12
pKa, MA 13.27 slow 0.96 1.04
WS, g L−1 (11.38) 20.55 skin 0.92 1.02
Kp, cm hr−1 NA
Log Kp NA

2 anti-(E)-methomyl
Ethanimidothioic acid, N
N-[[(methylamino) carbonyl] oxy]-methyl ester, (E)-
HN CAS No. 19928-35-9 fat 0.36 0.42
MW, g mol−1 162.21 brain 1.23 1.29
S N
O O Exp Kow @ pH 7.0 NA rapid 1.01 1.00
LogD @ pH 7.4 0.60 kidney 1.03 1.07
LogP 0.60 liver 0.96 1.12
pKa, MA 13.27 slow 0.96 1.04
WS, g L−1 (11.38) 20.55 skin 0.92 1.02

3 syn-(Z)-methomyl oxime
Ethanimidothioic acid, N
N-hydroxy-, methyl ester, (Z)-
S CAS No. 19125-12-3 fat 0.95 0.98
MW, g mol−1 105.16 brain 1.79 1.96
OH
N Exp Kow @ pH 7.0 NA rapid 1.18 1.13
LogD @ pH 7.4 1.12 kidney 1.23 1.30
LogP 1.12 liver 1.19 1.52
pKa, MA 11.26 slow 1.07 1.27
WS, g L−1 (16.69) 12.47 skin 1.21 1.31

4 anti-(E)-methomyl oxime
Ethanimidothioic acid, N
N-hydroxy-, methyl ester, (E)-
S CAS No. 19145-16-5 fat 0.95 0.98
MW, g mol−1 105.16 brain 1.79 1.96
N Exp Kow @ pH 7.0 NA rapid 1.18 1.13
LogD @ pH 7.4 1.12 kidney 1.23 1.30
OH
LogP 1.12 liver 1.19 1.52
pKa, MA 11.26 slow 1.07 1.27
WS, g L−1 (16.69) 12.47 skin 1.21 1.31
 Parameters for Carbamate Models 169

Table 19. (cont.)

Pesticide, pesticide ficiente

Partition coeffi
metabolite, and Physical and chemical
No.a chemical structureb propertiesc,d Tissue Rat Human

5 S-Methyl, N
N-methyl carbamic acid
Carbamothioic acid, methyl-, S-methyl ester
O CAS No. 22013-97-4 fat 0.24 0.31
MW, g mol−1 105.16 brain 1.11 1.14
S N Exp Kow @ pH 7.0 NA rapid 0.97 0.97
H LogD @ pH 7.4 0.30 kidney 0.99 1.02
LogP 0.30 liver 0.91 1.03
pKa, MA 12.77 slow 0.93 0.99
WS, g L−1 (41.17) 62.53 skin 0.85 0.95

6 Ethylium
Ethylium, 1-[(methylthio) imino]-
S CAS No. 61599-22-2 fat 1.13 1.15
N
MW, g mol−1 89.16 brain 1.95 2.14
Exp Kow @ pH 7.0 NA rapid 1.22 1.17
LogD @ pH 7.4 1.20 kidney 1.29 1.36
LogP 1.20 liver 1.25 1.63
pKa, MB 4.01 slow 1.11 1.33
WS, g L−1 (9.58) 11.81 skin 1.29 1.39

7 Carbon dioxide
O CAS No. 124-38-9 fat 0.53 0.58
MW, g mol−1 44.01 brain 1.40 1.50
O
Exp Kow @ pH 7.0 NA rapid 1.06 1.04
LogD @ pH 7.4 0.83 kidney 1.09 1.14
LogP 0.83 liver 1.03 1.25
pKa NA slow 0.99 1.11
WS, g L−1 (358.82) 25.72 skin 1.01 1.11

8 Acetonitrile
N CAS No. 75-05-8 fat 0.16 0.23
MW, g mol−1 41.05 brain 1.00 1.01
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −0.45 kidney 0.95 0.98
LogP −0.45 liver 0.87 0.95
pKa NA slow 0.91 0.94
WS, g L−1 (122.06) 312.45 skin 0.80 0.90
170 J.B. Knaak et al.

Table 19. (cont.)

Pesticide, pesticide ficiente

Partition coeffi
metabolite, and Physical and chemical
No.a chemical structureb propertiesc,d Tissue Rat Human

9 Hydrogen cyanide
N CAS No. 74-90-8 fat 0.15 0.23
MW, g mol−1 27.03 brain 1.02 1.03
Exp Kow @ pH 7.0 NA rapid 0.95 0.95
LogD @ pH 7.4 −0.25 kidney 0.95 0.98
LogP −0.25 liver 0.88 0.96
pKa NA slow 0.91 0.95
WS, g L−1 (89.68) 175.60 skin 0.81 0.90

10 Glutathione conjugate of acetonitrile

2-Amino-5-({2-[(carboxylmethyl)amino]-1-[(ethanimidoylthio)methyl]-2-
oxoethyl}amino)-5-oxopentanoic acid
O CAS No. NA fat 0.14 0.22
MW, g mol−1 348.38 brain 0.98 0.99
H2N
OH Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.0 −3.98 kidney 0.94 0.97
OH LogP −1.11 liver 0.86 0.94
pKa, MA, A 3.57 slow 0.91 0.93
O NH O 3.94
WS, g L−1 (3.56) 15877.38 skin 0.80 0.88
HN S NH

11 Glutamylcysteine conjugate of acetonitrile

2-Amino-5-{[1-carboxy-2-(ethanimidoylthio)ethyl}-5-oxopentanoic acid
O CAS No. NA fat 0.14 0.22
MW, g mol−1 291.33 brain 0.98 0.99
H2N
OH Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −3.67 kidney 0.94 0.97
LogP −0.92 liver 0.86 0.94
pKa, MA, A 3.15 slow 0.91 0.93
O NH 3.94
WS, g L−1 (4.3) 18780.45 skin 0.80 0.88
HN S OH

O
 Parameters for Carbamate Models 171

Table 19. (cont.)

Pesticide, pesticide ficiente

Partition coeffi
metabolite, and Physical and chemical
No.a chemical structureb propertiesc,d Tissue Rat Human

12 Cysteinylglycine conjugate of acetonitrile

{[2-Amino-3-(ethanimidoylthio)propanoyl]amino}acetic acid
NH CAS No. NA fat 0.14 0.22
MW, g mol−1 219.26 brain 0.98 0.99
S Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −3.62 kidney 0.94 0.97
NH2
LogP −0.85 liver 0.86 0.94
pKa, MA 3.61 slow 0.91 0.93
OH WS, g L−1 (22.28) 42875.65 skin 0.80 0.88
O N
H
O

13 Cysteine conjugate of acetonitrile

2-Amino-3-(ethanimidoylthio) propanoic acid
NH CAS No. NA fat 0.14 0.22
MW, g mol−1 162.21 brain 0.98 0.99
S Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −2.67 kidney 0.94 0.97
NH2
LogP −0.025 liver 0.86 0.94
pKa, MA 3.54 slow 0.91 0.93
O OH WS, g L−1 (25.95) 12745.72 skin 0.80 0.88

14 N-Sulfate-cysteine conjugate of acetonitrile

N
3-Mercapto-2-{[(1Z)-N-(sulfi
N finooxy)ethanimidoyl]amino}propanoic acid
O OH CAS No. NA fat 0.14 0.22
S MW, g mol−1 242.27 brain 0.98 0.99
O Exp Kow @ pH 7.0 NA rapid 0.94 0.94
N LogD @ pH 7.4 −4.73 kidney 0.94 0.97
LogP −0.083 liver 0.86 0.94
HN PKa, MA, A 2.15 slow 0.91 0.93
OH 3.24
WS, g L−1 (1000.0) 285646.80 skin 0.80 0.88
SH O

a
Parent chemical or metabolite number for cross-reference to Table 29 (Appendix B).
b–e
See footnotes in Table 14 (Appendix A).
172 J.B. Knaak et al.

Table 20. Chemical Structure, Physical and Chemical Properties, and Tissue Parti-
tion Coeffi
ficients for Oxamyl and the Resulting Metabolites.

Pesticide, pesticide ficiente

Partition coeffi
metabolite, and chemical Physical and chemical
No.a structureb propertiesc,d Tissue Rat Human

1 Oxamyl
Ethanimidothioic acid, 2-(dimethylamino)-N-[[(methylamino)]
N oxy]-oxo-, methyl
ester; CAS No. for Z isomer, 32817-80-4, is shown; CAS No. for E isomer,
32817-79-1.
O CAS No. 23135-22-0 fat 0.16 0.23
MW, g mol−1 219.26 brain 1.00 1.01
O N Exp Kow @ pH 7.0 NA rapid 0.94 0.94
H
S N LogD @ pH 7.4 −0.47 kidney 0.95 0.98
LogP −0.47 liver 0.87 0.95
pKa, MA 10.48 slow 0.91 0.94
O N WS, g L−1 (14.15) 87.52 skin 0.80 0.90
Kp, cm hr−1 NA
Log Kp NA

2 des-N-Oxamyl
N
Ethanimidothioic acid, 2-(methylamino)-N[[(methylamino) carbonyl] oxy]-2-oxo-,
methyl ester, (Z
Z isomer shown, no CAS No. for isomer)
S CAS No. 50917-40-3 fat 0.27 0.34
MW, g mol−1 205.24 brain 1.14 1.18
O O O
N Exp Kow @ pH 7.0 NA rapid 0.98 0.98
LogD @ pH 7.4 0.40 kidney 0.99 1.03
HN HN
LogP 0.40 liver 0.93 1.05
pKa, MA 9.28 slow 0.94 1.00
WS, g L−1 (5.18) 19.10 skin 0.87 0.97

3 Oxamyl oxime
Ethanimidothioic acid, 2-(dimethylamino)-N-hydroxy-2-oxo-,
N methyl ester; CAS No.
for Z isomer, 66344-33-0 is shown; CAS No. for E isomer, 66344-32-9.
S CAS No. 66344-33-0 fat 0.22 0.30
MW, g mol−1 162.21 brain 1.09 1.11
O OH
N Exp Kow @ pH 7.0 NA rapid 0.97 0.96
LogD @ pH 7.4 0.22 kidney 0.98 1.01
N
LogP 0.34 liver 0.91 1.01
pKa, MA 8.47 slow 0.93 0.98
WS, g/L (23.97) 43.38 skin 0.84 0.94

4 Oxamyl nitrile, DMCF

Carbonocyanidic amide, dimethyl-
CAS No. 16703-51-8 fat 0.15 0.23
N O MW, g mol−1 98.10 brain 0.98 0.99
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −0.93 kidney 0.94 0.97
LogP −0.93 liver 0.87 0.94
N
pKa, MB −2.15 slow 0.91 0.93
WS, g L−1 (147.30) 737.41 skin 0.79 0.89
 Parameters for Carbamate Models 173

Table 20. (cont.)

Pesticide, pesticide ficiente

Partition coeffi
metabolite, and chemical Physical and chemical
No.a structureb propertiesc,d Tissue Rat Human

5 des-N-Oxamyl
N oxime
Ethanimidothioic acid, N
N-hydroxy-2-(methylamino)-2-oxo-, methyl ester
O CAS No. 66157-67-3 fat 0.26 0.33
S MW, g mol−1 148.18 brain 1.12 1.16
HN Exp Kow @ pH 7.0 NA rapid 0.98 0.97
N LogD @ pH 7.4 0.35 kidney 0.99 1.03
OH LogP 0.38 liver 0.92 1.04
pKa, MA 9.63 slow 0.94 0.99
WS, g L−1 (15.42) 38.83 skin 0.86 0.96

6 Oxamyl oxime glucuronic acid

6-({[(1Z)-2-(Dimethylamino)-1-(methylthio)-2-oxoethylidene]amino}oxy)-3,4,5-
trihydroxytetrahydro-2H-pyran_2-carboxylic
H acid
O CAS No. NA fat 0.14 0.22
S MW, g mol−1 338.33 brain 0.98 0.99
N Exp Kow @ pH 7.0 NA rapid 0.94 0.94
N LogD @ pH 7.4 −3.03 kidney 0.94 0.97
O LogP 0.70 liver 0.86 0.94
OH pKa, MA 2.68 slow 0.91 0.93
O WS, g L−1 (1000.00) 2817.66 skin 0.79 0.88
HO
OH
O OH

7 Oxamyl acid (N,N-dimethyloxamic

N acid)
Acetic acid, (dimethylamino) oxo-
O OH CAS No. 32833-96-8 fat 0.14 0.22
MW, g mol−1 117.10 brain 0.98 0.99
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O N LogD @ pH 7.4 −4.98 kidney 0.94 0.97
LogP −1.28 liver 0.86 0.94
pKa, MA 3.13 slow 0.91 0.93
WS, g L−1 (1000.0) 1840664.00 skin 0.79 0.88

8 des-N-Oxamyl
N oxime glucuronic acid
3,4,5-Trihydroxy-6-({[(1Z)-2-(methylamino)-1-(methylthio)-2-oxoethylidene]
amino}oxy)tetrahydro-2H-pyran-2-carboxylic
H acid
O OH CAS No. NA fat 0.14 0.22
MW, g mol−1 324.31 brain 0.98 0.99
HO Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O HN
LogD @ pH 7.4 −2.99 kidney 0.94 0.97
N LogP 0.73 liver 0.86 0.94
HO O O
pKa, MA 2.68 slow 0.91 0.93
OH S WS, g L−1 (1000.0) 3096.82 skin 0.79 0.88
174 J.B. Knaak et al.

Table 20. (cont.)

Pesticide, pesticide ficiente

Partition coeffi
metabolite, and chemical Physical and chemical
No.a structureb propertiesc,d Tissue Rat Human

9 Glucuronide of N,N-dimethyloxamic
N acid
6-[2-(Dimethylamino)(oxo)acetoxy]-3,4,5-trihydroxytetrahydro-2H-pyran-2-
H
carboxylic acid
O CAS No. NA fat 0.14 0.22
MW, g mol−1 293.23 brain 0.98 0.99
O
OH N Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −4.65 kidney 0.94 0.97
O O
O LogP −0.93 liver 0.86 0.94
pKa, MA 2.64 slow 0.91 0.93
HO OH WS, g L−1 (1000.0) 125778.30 skin 0.79 0.88

OH

10 des-N-Oxamyl
N acid (N
N-methyloxamic acid)
Acetic acid, (methylamino) oxo-
O CAS No. 29262-58-6 fat 0.14 0.22
MW, g mol−1 103.08 brain 0.98 0.99
O
OH Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −4.99 kidney 0.94 0.97
HN
LogP −1.24 liver 0.86 0.94
pKa, MA 2.37 slow 0.91 0.93
WS, g L−1 (1000.0) 2090361.00 skin 0.79 0.88

11 Glucuronide of NN-methyloxamic acid

3,4,5-Trihydroxy-6-[2-(methylamino)(oxo)acetoxy]tetrahydro-2H-pyran-2-carboxylic
H
acid
OH O CAS No. NA fat 0.14 0.22
−1
MW, g mol 279.20 brain 0.98 0.99
HO
OH Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −4.61 kidney 0.94 0.97
O
HO LogP −0.89 liver 0.86 0.94
pKa, MA 2.63 slow 0.91 0.93
O O
WS, g L−1 (1000.0) 140047.00 skin 0.79 0.88

HN O

a
Parent chemical or metabolite number for cross-reference to Table 30 (Appendix B).
b–e
See footnotes in Table 14 (Appendix A).
 Parameters for Carbamate Models 175

Table 21. Chemical Structure, Physical and Chemical Properties, and Tissue Parti-
tion Coeffi
ficients for Pirimicarb and the Resulting Metabolites.

Pesticide, pesticide ficiente

Partition coeffi
metabolite, and chemical Physical and chemical
No.a structureb propertiesc,d Tissue Rat Human

1 Pirimicarb
Carbamic acid, dimethyl-, 2-(dimethylamino)-5,6-dimethyl-4-pyrimidinyl ester
N CAS No. 23103-98-2 fat 3.68 3.54
MW, g mol−1 238.29 brain 3.83 4.17
Exp Kow @ pH 7.0 NA rapid 1.78 1.58
O N N
LogD @ pH 7.4 1.70 kidney 1.96 2.05
LogP 1.70 liver 2.00 2.86
N O
pKa, MB 4.45 slow 1.49 2.04
WS, g L−1 (11.24) 0.96 skin 2.28 2.29
Kp, cm hr−1 NA
Log Kp NA

2 Hydroxymethyl pirimicarb
2-(Dimethylamino)-5,6-dimethylpyrimidin-4-yl (hydroxymethyl)methylcarbamate
N
CAS No. NA fat 0.61 0.66
MW, g mol−1 254.29 brain 1.48 1.60
Exp Kow @ pH 7.0 NA rapid 1.09 1.06
O N N
LogD @ pH 7.4 0.91 kidney 1.12 1.18
LogP 0.91 liver 1.07 1.31
N O 13.17 slow 1.01 1.15
pKa, MA
WS, g L−1 (65.65) 3.74 skin 1.05 1.15
HO

3 Pirimicarb phenol
4(1H)-Pyrimidinone,
H 2-(dimethylamino), 5,6-dimethyl-
N CAS No. 40778-16-3 fat 0.15 0.22
MW, g mol−1 167.21 brain 0.98 0.99
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
N N
LogD @ pH 7.4 −0.99 kidney 0.94 0.97
LogP 1.20 liver 0.87 0.94
HO
pKa, MA 7.58 slow 0.91 0.93
WS, g L−1 (2.8) 444.05 skin 0.79 0.89

4 Demethyl pirimicarb phenol

4(1H)-Pyrimidinone,
H 5,6-dimethyl-2-(methylamino)-
NH CAS No. 78195-30-9 fat 0.15 0.22
MW, g mol−1 153.18 brain 0.98 0.99
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
N N
LogD @ pH 7.4 −1.42 kidney 0.94 0.97
LogP 0.40 liver 0.86 0.94
HO
pKa, MA 7.66 slow 0.91 0.93
WS, g L−1 (8.74) 1198.70 skin 0.79 0.88
176 J.B. Knaak et al.

Table 21. (cont.)

e
Pesticide, pesticide Partition coefficient
fi
metabolite, and chemical Physical and chemical
No.a structureb propertiesc,d Tissue Rat Human

5 Amino pirimicarb phenol

4(1H)-Pyrimidinone,
H 2-amino-5,6-dimethyl-
NH2 CAS No. 3977-23-9 fat 0.14 0.22
MW, g mol−1 139.16 brain 0.98 0.99
N N Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −2.02 kidney 0.94 0.97
HO LogP −0.09 liver 0.86 0.94
pKa, MA 7.54 slow 0.91 0.93
WS, g L−1 (16.72) 4482.52 skin 0.79 0.88

6 Hydroxymethyl demethyl pirimicarb phenol

4 (1H)-Pyrimidinone,
H 6-(hydroxymethyl)-5-methyl-2-(methylamino)-
NH CAS No. 78195-34-3 fat 0.14 0.22
MW, g mol−1 169.18 brain 0.98 0.99
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
N N
LogD @ pH 7.4 −2.31 kidney 0.94 0.97
OH LogP −1.24 liver 0.86 0.94
HO
pKa, MA 7.33 slow 0.91 0.93
WS, g L−1 (46.15) 5826.93 skin 0.79 0.88

7 Pirimicarb phenyl glucuronic acid

6-{[2-(Dimethylamino)-5,6-dimethylpyrimidin-4-yl]oxy}-3,4,5-trihydroxytetrahydro-
2H-pyran-2-carboxylic
H acid
CAS No. NA fat 0.14 0.22
N N MW, g mol−1 343.34 brain 0.98 0.99
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
N LogD @ pH 7.4 −3.77 kidney 0.94 0.97
OH LogP −0.15 liver 0.86 0.94
O OH pKa, MA 2.65 slow 0.91 0.93
WS, g L−1 (16.60) 11268.07 skin 0.79 0.88
O
OH

HO O

8 Hydroxymethyl pirimicarb phenol

4 (1H)-Pyrimidinone,
H 2-(dimethylamino)-6-(hydroxymethyl)-5-methyl-
N CAS No. 244247-04-9 fat 0.14 0.22
MW, g mol−1 183.21 brain 0.98 0.99
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
N N
LogD @ pH 7.4 −2.0 kidney 0.94 0.97
OH LogP −0.44 liver 0.86 0.94
HO
pKa, MA 7.15 slow 0.91 0.93
WS, g L−1 (14.1) 2711.32 skin 0.79 0.88

a
Parent chemical or metabolite number for cross-reference to Table 31 (Appendix B).
b–e
See footnotes in Table 14 (Appendix A).
 Parameters for Carbamate Models 177

Table 22. Chemical Structure, Physical and Chemical Properties, and Tissue Parti-
tion Coeffi
ficients for Propoxur and the Resulting Metabolites.

Pesticide, pesticide ficiente

Partition coeffi
metabolite, and chemical Physical and chemical
No.a structureb propertiesc,d Tissue Rat Human

1 Propoxur
Phenol, 2-(1-methylethoxy)-, methylcarbamate
CAS No. 114-26-1 fat 2.88 2.80
HN O MW, g mol−1 209.24 brain 3.29 3.61
Exp Kow @ pH 7.0 NA rapid 1.60 1.47
O LogD @ pH 7.4 1.60 kidney 1.77 1.86
LogP 1.60 liver 1.79 2.52
pKa, MA 12.28 slow 1.38 1.85
O
WS, g L−1 (0.71) 1.69 skin 2.00 2.04
Kp, cm hr−1 NA
Log Kp NA

2 2-Isopropoxy phenol
Phenol, 2-(1-methylethoxy)-
CAS No. 4812-20-8 fat 9.17 8.60
MW, g mol−1 152.19 brain 6.87 6.94
O
Exp Kow @ pH 7.0 NA rapid 2.08 2.13
HO LogD @ pH 7.4 2.06 kidney 3.04 3.00
LogP 2.07 liver 3.21 4.53
pKa, MA 9.99 slow 2.12 3.01
WS, g L−1 (2.77) 1.29 skin 3.86 3.51

3 2-OH Phenyl methylcarbamate

2-Hydroxy phenyl methylcarbamate
CAS No. 10309-97-4 fat 0.14 0.22
HN O MW, g mol−1 167.16 brain 0.98 0.99
OH Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O LogD @ pH 7.4 0.19 kidney 0.94 0.97
LogP 0.22 liver 0.86 0.94
pKa, MA 8.69 slow 0.91 0.93
WS, g L−1 (10.69) 43.6 skin 0.79 0.88

4 5-OH Propoxur
1,3-Benzenediol, 4-(1-methylethoxy)-, 3-(methylcarbamate)
CAS No. 13200-88-9 fat 0.66 0.71
HN O MW, g mol−1 225.24 brain 1.54 1.65
Exp Kow @ pH 7.0 NA rapid 1.10 1.07
O OH LogD @ pH 7.4 0.95 kidney 1.14 1.20
LogP 0.95 liver 1.09 1.34
pKa, MA 9.93 slow 1.02 1.17
O
WS, g L−1 (2.02) 4.99 skin 1.08 1.18
178 J.B. Knaak et al.

Table 22. (cont.)

Pesticide, pesticide ficiente

Partition coeffi
metabolite, and chemical Physical and chemical
No.a structureb propertiesc,d Tissue Rat Human

5 2-Isopropoxy 5-OH phenol

4-Isopropoxybenzene-1,3-diol
CAS No. NA fat 1.26 1.28
MW, g mol−1 168.19 brain 2.06 2.77
O Exp Kow @ pH 7.0 NA rapid 1.26 1.20
HO LogD @ pH 7.4 1.25 kidney 1.33 1.40
LogP 1.26 liver 1.30 1.71
pKa, MA 9.69 slow 1.13 1.38
WS, g L−1 (9.23) 5.37 skin 1.35 1.45
OH

6 4-OH Propoxur
Carbamic acid, methyl-, 4-hydroxy-2-isopropoxyphenyl ester
CAS No. 17595-59-4 fat 0.59 0.64
HN O MW, g mol−1 225.24 brain 1.46 1.57
Exp Kow @ pH 7.0 NA rapid 1.08 1.05
O LogD @ pH 7.4 0.89 kidney 1.11 1.17
LogP 0.89 liver 1.06 1.29
pKa, MA 9.40 slow 1.01 1.14
O OH
WS, g L−1 (2.14) 5.61 skin 1.04 1.14

7 2-Isopropoxy 4-OH phenol

Hydroquinone, isopropoxy-
CAS No. 20042-92-6 fat 1.60 1.59
MW, g mol−1 168.19 brain 2.33 2.57
O Exp Kow @ pH 7.0 NA rapid 1.34 1.26
HO LogD @ pH 7.4 1.35 kidney 1.42 1.51
LogP 1.35 liver 1.40 1.89
pKa, MA 10.47 slow 1.18 1.49
OH WS, g L−1 (8.51) 4.41 skin 1.49 1.58

8 N-Hydroxymethyl propoxur
Carbamic acid, (hydroxymethyl)-, 2-(1-methylethoxy) phenyl ester
HO CAS No. 10310-16-4 fat 0.50 0.56
MW, g mol−1 225.24 brain 1.38 1.46
HN O Exp Kow @ pH 7.0 NA rapid 1.05 1.03
O
LogD @ pH 7.4 0.80 kidney 1.08 1.13
O LogP 0.80 liver 1.02 1.23
pKa, A 13.59 slow 0.99 1.10
WS, g L−1 (4.22) 6.70 skin 0.99 1.10
 Parameters for Carbamate Models 179

Table 22. (cont.)

Pesticide, pesticide ficiente

Partition coeffi
metabolite, and chemical Physical and chemical
No.a structureb propertiesc,d Tissue Rat Human

9 2-Isopropoxy phenyl sulfuric acid

Phenol, 2-(1-methylethoxy)-, hydrogen sulfate
OH CAS No. 152242-55-2 fat 0.14 0.22
O S O MW, g mol−1 232.25 brain 0.98 0.99
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O LogD @ pH 7.4 −1.75 kidney 0.94 0.97
LogP 1.75 liver 0.86 0.94
O pKa, MA −3.97 slow 0.91 0.93
WS, g L−1 (1000.0) 923.43 skin 0.79 0.88

10 2-Isopropoxy phenyl glucuronic acid

β-d-glucopyranosiduronic acid, 2-(1-methylethoxy) phenyl
OH O CAS No. 152242-54-1 fat 0.14 0.22
−1
HO MW, g mol 328.31 brain 0.98 1.09
OH Exp Kow @ pH 7.0 NA rapid 0.94 0.96
O LogD @ pH 7.4 −4.10 kidney 0.94 1.00
HO O LogP −0.39 liver 0.86 1.00
O pKa, MA 2.79 slow 0.91 0.97
WS, g L−1 (1000.0) 26524.48 skin 0.79 0.93

11 2-Hydroxyphenyl methylcarbamate sulfuric acid

2-(Sulfooxy) phenyl methylcarbamate
CAS No. NA fat 0.14 0.22
HN O MW, g mol−1 247.23 brain 0.98 0.99
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O LogD @ pH 7.4 −3.6 kidney 0.94 0.97
O LogP −0.1 liver 0.86 0.94
O
S pKa, MA −4.25 slow 0.91 0.93
O WS, g L−1 (1000.0) 29084.9 skin 0.79 0.88
HO

12 2-Hydroxyphenyl methylcarbamate glucuronic acid

3,4,5-Trihydroxy-6-{2-[(methylcarbamoyl)oxy][phenoxy}
tetrahydro-2H-pyran-2-carboxylic
H acid
HO O CAS No. NA fat 0.14 0.22
MW, g mol−1 343.29 brain 0.98 0.99
OH Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O LogD @ pH 7.4 −5.95 kidney 0.94 0.97
HN O LogP −2.23 liver 0.86 0.94
O OH pKa, MA 2.77 slow 0.91 0.93
−1
O OH WS, g L (1000.0) 819992.6 skin 0.79 0.88
180 J.B. Knaak et al.

Table 22. (cont.)

Pesticide, pesticide ficiente

Partition coeffi
metabolite, and chemical Physical and chemical
No.a structureb propertiesc,d Tissue Rat Human

13 5-OH Propoxur sulfuric acid

2-Isopropoxy-5-(sulfooxy) phenyl methylcarbamate
CAS No. NA fat 0.14 0.22
HN MW, g mol−1 305.30 brain 0.98 0.99
O O Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O LogD @ pH 7.4 −2.73 kidney 0.94 0.97
LogP 0.77 liver 0.86 0.94
pKa, MA −4.09 slow 0.91 0.93
WS, g L−1 (1000.0) 2452.24 skin 0.79 0.88
O
O
S
O
HO

14 5-OH Propoxur glucuronic acid

4-Isopropoxy-3-[(methylcarbamoyl)oxy]phenyl hexopyranosiduronic acid
CAS No. NA fat 0.14 0.22
HN O MW, g mol−1 401.37 brain 0.98 0.99
OH
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O O O LogD @ pH 7.4 −5.08 kidney 0.94 0.97
O
LogP −1.37 liver 0.86 0.94
O HO OH pKa, MA 2.77 slow 0.91 0.93
WS, g L−1 (1000.0) 65419.22 skin 0.79 0.88
OH

15 2-Isopropoxy 5-OH phenyl sulfuric acid

5-Hydroxy-2-isopropoxyphenyl hydrogen sulfate
OH CAS No. NA fat 0.14 0.22
O S O MW, g mol−1 248.25 brain 0.98 0.99
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O OH LogD @ pH 7.4 −2.4 kidney 0.94 0.97
LogP 1.10 liver 0.86 0.94
O pKa, MA −4.21 slow 0.91 0.93
WS, g L−1 (1000.0) 2709.80 skin 0.79 0.88

16 2-Isopropoxy 5-OH phenyl glucuronic acid

3,5,5-Trihydroxy-6-(5-hydroxy-2-isopropoxyphenoxy)
tetrahydro-2H-pyran-2-carboxylic
H acid
OH O CAS No. NA fat 0.14 0.22
HO MW, g mol−1 344.32 brain 0.98 0.99
OH Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O LogD @ pH 7.4 −4.75 kidney 0.94 0.97
HO O LogP −1.03 liver 0.86 0.94
O pKa, MA 2.76 slow 0.91 0.93
WS, g L−1 (1000.0) 76371.80 skin 0.79 0.88

OH
 Parameters for Carbamate Models 181

Table 22. (cont.)

Pesticide, pesticide ficiente

Partition coeffi
metabolite, and chemical Physical and chemical
No.a structureb propertiesc,d Tissue Rat Human

17 4-OH Propoxur sulfuric acid

2-Isopropoxy-4-(sulfooxy) phenyl methylcarbamate
CAS No. NA fat 0.14 0.22
HN MW, g mol−1 305.31 brain 0.98 0.99
O O Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O LogD @ pH 7.4 −2.73 kidney 0.94 0.97
LogP 0.77 liver 0.86 0.94
O pKa, MA −4.24 slow 0.91 0.93
WS, g L−1 (1000.0) 2452.24 skin 0.79 0.88
HO S O
O

18 4-OH propoxur glucuronic acid

2,3,4-Trihydroxy-5-{3-isopropoxy-4-[(methylcarbamoyl)oxy]phenoxy}
cyclohexanecarboxylic acid
CAS No. NA fat 0.14 0.22
HN O MW, g mol−1 401.37 brain 0.98 0.99
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O LogD @ pH 7.4 −5.08 kidney 0.94 0.97
LogP −1.37 liver 0.86 0.94
pKa, MA 2.77 slow 0.91 0.93
O O
WS, g L−1 (1000.0) 65419.22 skin 0.79 0.88
HO
O
OH
HO
OH O

19 2-Isopropoxy 4-OH phenyl sulfuric acid

4-Hydroxy-2-isopropoxyphenyl hydrogen sulfate
OH CAS No. NA fat 0.14 0.22
O S O MW, g mol−1 248.25 brain 0.98 0.99
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O LogD @ pH 7.4 −2.47 kidney 0.94 0.97
LogP 1.04 liver 0.86 0.94
O OH pKa, MA −4.24 slow 0.91 0.93
WS, g L−1 (1000.0) 3109.80 skin 0.79 0.88
182 J.B. Knaak et al.

Table 22. (cont.)

Pesticide, pesticide ficiente

Partition coeffi
metabolite, and chemical Physical and chemical
No.a structureb propertiesc,d Tissue Rat Human

20 2-Isopropoxy 4-OH phenyl glucuronic acid

3,4,5-Trihydroxy-6-(4-hydroxy-2-isopropoxyphenoxy)
tetrahydro-2H-pyran-2-carboxylic
H acid
OH O CAS No. NA fat 0.14 0.22
HO MW, g mol−1 344.32 brain 0.98 0.99
OH Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O LogD @ pH 7.4 −4.81 kidney 0.94 0.97
HO O LogP −1.09 liver 0.86 0.94
O pKa, MA 2.78 slow 0.91 0.93
WS, g L−1 (1000.0) 85935.58 skin 0.79 0.88

OH

21 N-Hydroxymethyl propoxur sulfuric acid

2-Isopropoxyphenyl [(sulfooxy)methyl]carbamate
O CAS No. NA fat 0.14 0.22
−1
O S OH MW, g mol 305.31 brain 0.98 0.99
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
HN O
LogD @ pH 7.4 −2.43 kidney 0.94 0.97
O
LogP 1.07 liver 0.86 0.94
O pKa, MA −3.88 slow 0.91 0.93
O WS, g L−1 (1000.0) 1359.5 skin 0.79 0.88

22 N-Hydroxymethyl propoxur glucuronic acid

{[(2-Isopropoxyphenoxy)carbonyl]amino}methyl hexopyranosiduronic acid
CAS No. NA fat 0.14 0.22
OH O
MW, g mol−1 401.37 brain 0.98 0.99
H Exp Kow @ pH 7.0 NA rapid 0.94 0.94
O O N O
O LogD @ pH 7.4 −4.93 kidney 0.94 0.97
O LogP −1.22 liver 0.86 0.94
HO OH pKa, MA 2.84 slow 0.91 0.93
OH WS, g L−1 (1000.0) 48729.4 skin 0.79 0.88

a
Parent chemical or metabolite number for cross-reference to Table 32 (Appendix B).
b–e
See footnotes in Table 14 (Appendix A).
 Parameters for Carbamate Models 183

Table 23. Chemical Structure, Physical and Chemical Properties, and Tissue Parti-
tion Coeffi
ficients for Thiodicarb and the Resulting Metabolites.

Pesticide, pesticide ficiente

Partition coeffi
metabolite, and chemical Physical and chemical
No.a structureb propertiesc,d Tissue Rat Human

1 Thiodicarb
Ethanimidothioic acid, N-
N N’-[thiobis[(methylimino) carbonyloxy]] bis-, dimethyl
ester
CAS No. 59669-26-0 fat 2.38 2.33
MW, g mol−1 354.47 brain 2.93 3.23
N S
Exp Kow @ pH 7.0 NA rapid 1.51 1.39
O O LogD @ pH 7.4 1.52 kidney 1.64 1.73
LogP 1.52 liver 1.64 2.29
N pKa, MB −2.09 slow 1.31 1.72
S N S
WS, g L−1 (0.19) 0.29 skin 1.81 1.88
N O Kp, cm hr−1 NA
Log Kp NA
O

2 syn-(Z)-Methomyl
Ethanimidothioic acid, N
N-[[(methylamino)carbonyl]oxy]- methyl ester, (Z)-
O CAS No. 19928-37-1 fat 0.36 0.42
MW, g mol−1 162.21 brain 1.23 1.29
O N Exp Kow @ pH 7.0 NA rapid 1.01 1.00
H
S N LogD @ pH 7.4 0.60 kidney 1.03 1.07
LogP 0.60 liver 0.96 1.12
pKa, MA 13.27 slow 0.96 1.04
WS, g L−1 (11.38) 20.55 skin 0.92 1.02

3 anti-(E)-Methomyl
Ethanimidothioic acid, N
N-[[(methylamino)carbonyl]oxy]- methyl ester, (E)-
HN CAS No. 19928-35-9 fat 0.36 0.42
MW, g mol−1 162.21 brain 1.23 1.29
S N
O O Exp Kow @ pH 7.0 NA rapid 1.01 1.00
LogD @ pH 7.4 0.60 kidney 1.03 1.07
LogP 0.60 liver 0.96 1.12
pKa, MA 13.27 slow 0.96 1.04
WS, g L−1 (11.38) 20.55 skin 0.92 1.02

4 ssyn-(Z)-Methomyl oxime
Ethanimidothioic acid, N
N-hydroxy-, methyl ester, (Z)-
S CAS No. 19125-12-3 fat 0.95 0.98
MW, g mol−1 105.16 brain 1.79 1.96
OH
N Exp Kow @ pH 7.0 NA rapid 1.18 1.13
LogD @ pH 7.4 1.12 kidney 1.23 1.30
LogP 1.12 liver 1.19 1.52
pKa, MA 11.26 slow 1.07 1.27
WS, g L−1 (16.69) 12.47 skin 1.21 1.31
184 J.B. Knaak et al.

Table 23. (cont.)

Pesticide, pesticide ficiente

Partition coeffi
metabolite, and chemical Physical and chemical
No.a structureb propertiesc,d Tissue Rat Human

5 anti-(E)-Methomyl oxime
Ethanimidothioic acid, N-hydroxy-, methyl ester, (E)-
S CAS No. 19145-16-5 fat 0.95 0.98
MW, g mol−1 105.16 brain 1.79 1.96
N Exp Kow @ pH 7.0 NA rapid 1.18 1.13
LogD @ pH 7.4 1.12 kidney 1.23 1.30
OH
LogP 1.12 liver 1.19 1.52
pKa, MA 11.26 slow 1.07 1.27
WS, g L−1 (16.69) 12.47 skin 1.21 1.31

6 S-Methyl, N
N-methyl carbamic acid
Carbamothioic acid, methyl-, S-methyl ester
O CAS No. 22013-97-4 fat 0.24 0.31
MW, g mol−1 105.16 brain 1.11 1.14
S N Exp Kow @ pH 7.0 NA rapid 0.97 0.97
H LogD @ pH 7.4 0.30 kidney 0.99 1.02
LogP 0.30 liver 0.91 1.03
pKa, MA 12.77 slow 0.93 0.99
WS, g L−1 (41.17) 62.53 skin 0.85 0.95

7 Ethylium
Ethylium, 1-[(methylthio) imino]-
S CAS No. 61599-22-2 fat 1.13 1.15
N
MW, g mol−1 89.16 brain 1.95 2.14
Exp Kow @ pH 7.0 NA rapid 1.22 1.17
LogD @ pH 7.4 1.20 kidney 1.29 1.36
LogP 1.20 liver 1.25 1.63
pKa, MB 4.01 slow 1.11 1.33
WS, g L−1 (9.58) 11.81 skin 1.29 1.39

8 Carbon dioxide
O CAS No. 124-38-9 fat 0.53 0.58
MW, g mol−1 44.01 brain 1.40 1.50
O
Exp Kow @ pH 7.0 NA rapid 1.06 1.04
LogD @ pH 7.4 0.83 kidney 1.09 1.14
LogP 0.83 liver 1.03 1.25
pKa NA slow 0.99 1.11
WS, g L−1 (358.82) 25.72 skin 1.01 1.11
 Parameters for Carbamate Models 185

Table 23. (cont.)

Pesticide, pesticide ficiente

Partition coeffi
metabolite, and chemical Physical and chemical
No.a structureb propertiesc,d Tissue Rat Human

9 Acetonitrile
N CAS No. 75-05-8 fat 0.16 0.23
MW, g mol−1 41.05 brain 1.00 1.01
Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −0.45 kidney 0.95 0.98
LogP −0.45 liver 0.87 0.95
pKa NA slow 0.91 0.94
WS, g L−1 (122.06) 312.45 skin 0.80 0.90

10 Hydrogen cyanide
N CAS No. 74-90-8 fat 0.15 0.23
MW, g mol−1 27.03 brain 1.02 1.03
Exp Kow @ pH 7.0 NA rapid 0.95 0.95
LogD @ pH 7.4 −0.25 kidney 0.95 0.98
LogP −0.25 liver 0.88 0.96
pKa NA slow 0.91 0.95
WS, g L−1 (89.68) 175.60 skin 0.81 0.90

11 Glutathione conjugate of acetonitrile

2-Amino-5-({2-[(carboxylmethyl)amino]-1-[(ethanimidoylthio)methyl]-2-
oxoethyl}amino)-5-oxopenanoic acid
O CAS No. NA fat 0.14 0.22
−1
MW, g mol 348.38 brain 0.98 0.99
H2N
OH Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −3.98 kidney 0.94 0.97
OH LogP −1.11 liver 0.86 0.94
pKa, MA, A 3.57 slow 0.91 0.93
O NH O 3.94
WS, g L−1 (3.56) 15877.38 skin 0.80 0.88
HN S NH

12 Glutamylcysteine conjugate of acetonitrile

2-Amino-5-{[1-carboxy-2-(ethanimidoylthio)ethyl}-5-oxopentanoic acid
O CAS No. NA fat 0.14 0.22
MW, g mol−1 291.33 brain 0.98 0.99
H2N
OH Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −3.67 kidney 0.94 0.97
LogP −0.92 liver 0.86 0.94
pKa, MA, A 3.15 slow 0.91 0.93
O NH 3.94
WS, g L−1 (4.3) 18780.45 skin 0.79 0.88
HN S OH

O
186 J.B. Knaak et al.

Table 23. (cont.)

Pesticide, pesticide ficiente

Partition coeffi
metabolite, and chemical Physical and chemical
No.a structureb propertiesc,d Tissue Rat Human

13 Cysteinylglycine conjugate of acetonitrile

{[2-Amino-3-(ethanimidoylthio)propanoyl]amino}acetic acid
NH CAS No. NA fat 0.14 0.22
MW, g mol−1 219.26 brain 0.98 0.99
S Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −3.62 kidney 0.94 0.97
NH2
LogP −0.85 liver 0.86 0.94
pKa, MA 3.61 slow 0.91 0.93
OH
O N WS, g L−1 (22.28) 42875.65 skin 0.80 0.88
H
O

14 Cysteine conjugate of acetonitrile

2-Amino-3-(ethanimidoylthio)propanoic acid
NH CAS No. NA fat 0.14 0.22
MW, g mol−1 162.21 brain 0.98 0.99
S Exp Kow @ pH 7.0 NA rapid 0.94 0.94
LogD @ pH 7.4 −2.67 kidney 0.94 0.97
NH2
LogP −0.025 liver 0.86 0.94
pKa, MA 3.54 slow 0.91 0.93
O OH WS, g L−1 (25.95) 12745.72 skin 0.80 0.88

15 N-Sulfate-cysteine conjugate of acetonitrile

N
3-Mercapto-2-{[(1Z)-N-(sulfi
N finooxy)ethanimidoyl]amino}propanoic acid
O OH CAS No. NA fat 0.14 0.22
S MW, g mol−1 242.27 brain 0.98 0.99
O Exp Kow @ pH 7.0 NA rapid 0.94 0.94
N LogD @ pH 7.4 −4.73 kidney 0.94 0.97
LogP −0.083 liver 0.86 0.94
HN pKa, MA, A 2.15 slow 0.91 0.93
OH 3.24
WS, g L−1 (1000.0) 285646.80 skin 0.80 0.88
SH O

a
Parent chemical or metabolite number for cross-reference to Table 33 (Appendix B).
b–e
See footnotes in Table 14 (Appendix A).
 Parameters for Carbamate Models 187

Appendix B. Metabolic Pathways and Preliminary Metabolic

Rate Constants (Vmax, Km) for the Metabolism of Parent
Carbamates and Metabolites.

Table 24. Biotransformation and Elimination Paths of Aldicarb and the Resulting
Metabolites and Preliminary Liver Vmax and Km Values.a

No.b Initiating pesticide/metabolitec Resulting metabolite

1 Aldicarb
Aldicarb oxime
Aldicarb sulfoxide
Aldicarb nitrile
Aldicarb eliminated

2 Aldicarb sulfoxide
Aldicarb oxime sulfoxide
Aldicarb sulfone
Aldicarb nitrile sulfoxide
Aldicarb sulfoxide eliminated

3 Aldicarb sulfone
Aldicarb oxime sulfone
Aldicarb nitrile sulfone
Aldicarb sulfone eliminated

4 Aldicarb oxime
Aldicarb aldehyde
Aldicarb oxime eliminated

5 Aldicarb oxime sulfoxide

Aldicarb aldehyde sulfoxide
Aldicarb oxime sulfoxide eliminated

6 Aldicarb oxime sulfone

Aldicarb aldehyde sulfone
Aldicarb oxime sulfone eliminated

7 Aldicarb nitrile
Aldicarb nitrile eliminated

8 Aldicarb nitrile sulfoxide

Aldicarb nitrile sulfoxide eliminated

9 Aldicarb nitrile sulfone

Aldicarb nitrile sulfone eliminated
188 J.B. Knaak et al.

Table 24. (cont.)

No.b Initiating pesticide/metabolitec Resulting metabolite

10 Aldicarb aldehyde
Aldicarb alcohol
Aldicarb acid

11 Aldicarb aldehyde sulfoxide

Aldicarb alcohol sulfoxide
Aldicarb acid sulfoxide

12 Aldicarb aldehyde sulfone

Aldicarb alcohol sulfone
Aldicarb acid sulfone

13 Aldicarb alcohol
Aldicarb alcohol eliminated

14 Aldicarb alcohol sulfoxide

Aldicarb alcohol sulfoxide eliminated

15 Aldicarb alcohol sulfone

Aldicarb alcohol sulfone eliminated

16 Aldicarb acid
Aldicarb acid eliminated

17 Aldicarb acid sulfoxide

Aldicarb acid sulfoxide eliminated

18 Aldicarb acid sulfone

Aldicarb acid sulfone eliminated
a
Preliminary estimates of liver Vmax (μmol hr−1kg−1 bw, unscaled) and Km (μM) are equal
to 10.0.
b
Parent chemical or metabolite number for cross-reference in Table 14 (Appendix A).
c
Refer to Table 14 for structure based on No.
 Parameters for Carbamate Models 189

Table 25. Biotransformation and Elimination Paths of Carbaryl and the Resulting
Metabolites and Preliminary Liver Vmax and Km Valuesa.

No.b Initiating pesticide/metabolitec Resulting metabolite

1 Carbaryl
Naphthol
4-OH Carbaryl
5-OH Carbaryl
3,4-Dihydrodihydroxy carbaryl
5,6-Dihydro-dihydroxy carbaryl
Hydroxymethyl carbaryl

2 4-OH carbaryl
4-OH Naphthol
4-OH Carbaryl sulfuric acid
4-OH Carbaryl glucuronic acid
4-OH Carbaryl eliminated

3 5-OH carbaryl
5-OH Naphthol
5-OH Carbaryl sulfuric acid
5-OH Carbaryl glucuronic acid
5-OH Carbaryl eliminated

4 3,4-Dihydrodihydroxy carbaryl
3,4-Dihydrodihydroxy naphthol
3,4-Dihydrodihydroxy carbaryl
sulfuric acid
3,4-Dihydrodihydroxy carbaryl
glucuronic acid
3,4-Dihydrodihydroxy carbaryl
eliminated

5 5,6-Dihydrodihydroxy carbaryl
5,6-Dihydrodihydroxy naphthol
5,6-Dihydrodihydroxy carbaryl
sulfuric acid
5,6-Dihydrodihydroxy carbaryl
glucuronic acid
5,6-Dihydrodihydroxy carbaryl
eliminated

6 Hydroxymethyl carbaryl
1-Naphthol
Hydroxymethyl carbaryl sulfuric acid
Hydroxymethyl carbaryl glucuronic
acid
Hydroxymethyl carbaryl eliminated
190 J.B. Knaak et al.

Table 25. (cont.)

No.b Initiating pesticide/metabolitec Resulting metabolite

7 Naphthol
2-OH Naphthol
4-OH Naphthol
5-OH Naphthol
3,4-Dihydrodihydroxy naphthol
5,6-Dihydrodihydroxy naphthol
Naphthyl sulfuric acid
Naphthyl glucuronic acid
Naphthol eliminated

8 1-Hydroxy-2-naphthol
2-OH Naphthyl sulfuric acid
2-OH Naphthyl glucuronic acid
2-OH Naphthol eliminated

9 4-OH naphthol
4-OH Naphthyl sulfuric acid
4-OH Naphthyl glucuronic acid
4-OH Naphthol eliminated

10 5-OH naphthol
5-OH Naphthyl sulfuric acid
5-OH Naphthyl glucuronic acid
5-OH Naphthol eliminated

11 3,4-Dihydrodihydroxy naphthol
3,4-Dihydrodihydroxy naphthyl
sulfuric acid
3,4-Dihydrodihydroxy naphthyl
glucuronic acid
3,4-Dihydrodihydroxy naphthol
eliminated

12 5,6-Dihydrodihydroxy naphthol
5,6-Dihydrodihydroxy naphthyl
sulfuric acid
5,6-Dihydrodihydroxy naphthyl
glucuronic acid
5,6-Dihydrodihydroxy naphthol
eliminated

13 Naphthyl sulfuric acid

Naphthyl sulfuric acid eliminated

14 Naphthyl glucuronic acid

Naphthyl glucuronic acid eliminated
 Parameters for Carbamate Models 191

Table 25. (cont.)

No.b Initiating pesticide/metabolitec Resulting metabolite

15 2-OH naphthyl sulfuric acid
2-OH naphthyl sulfuric acid
eliminated

16 2-OH naphthyl glucuronic acid

2-OH naphthyl glucuronic acid
eliminated

17 4-OH naphthyl sulfuric acid

4-OH naphthyl sulfuric acid eliminated

18 4-OH naphthyl glucuronic acid

4-OH naphthyl glucuronic acid
eliminated

19 5-OH naphthyl sulfuric acid

5-OH naphthyl sulfuric acid eliminated

20 5-OH naphthyl glucuronic acid

5-OH naphthyl glucuronic acid
eliminated

21 3,4-Dihydrodihydroxy naphthyl sulfuric acid

3,4-Dihydrodihydroxy naphthyl sulfuric
acid eliminated

22 3,4-Dihydrodihydroxy naphthyl glucuronic acid

3,4-Dihydrodihydroxy naphthyl
glucuronic acid eliminated

23 5,6-Dihydrodihydroxy naphthyl sulfuric acid

5,6-Dihydrodihydroxy naphthyl sulfuric
acid eliminated

24 5,6-Dihydrodihydroxy naphthyl glucuronic acid

5,6-Dihyrodihydroxy naphthyl
glucuronic acid eliminated

25 4-OH carbaryl sulfuric acid

4-OH carbaryl sulfuric acid eliminated

26 4-OH carbaryl glucuronic acid

4-OH carbaryl glucuronic acid
eliminated
192 J.B. Knaak et al.

Table 25. (cont.)

No.b Initiating pesticide/metabolitec Resulting metabolite

27 5-OH carbaryl sulfuric acid

5-OH carbaryl sulfuric acid eliminated

28 5-OH carbaryl glucuronic acid

5-OH carbaryl glucuronic acid
eliminated

29 3,4-Dihydrodihydroxy carbaryl sulfuric acid

3,4-Dihydrodihydroxy carbaryl sulfuric
acid eliminated

30 3,4-Dihydrodihydroxy carbaryl glucuronic acid

3,4-Dihydrodihydroxy carbaryl
glucuronic acid eliminated

31 5,6-Dihydrodihydroxy carbaryl sulfuric acid

5,6-Dihydrodihydroxy carbaryl sulfuric
acid eliminated

32 5,6-Dihydrodihydroxy carbaryl glucuronic acid

5,6-Dihydrodihydroxy carbaryl
glucuronic acid eliminated

33 Hydroxymethyl carbaryl sulfuric acid

Hydroxymethyl carbaryl sulfuric acid
eliminated

34 Hydroxymethyl carbaryl glucuronic acid

Hydroxymethyl carbaryl glucuronic
acid eliminated
a
Preliminary estimates of liver Vmax (μmol hr−1kg−1 bw, unscaled) and Km (μM) are equal
to 10.0. Additional published values can be found in Table 34 (Appendix B).
b
Parent chemical or metabolite number for cross-reference in Table 15 (Appendix A).
c
Refer to Table 15 for structure based on No.
 Parameters for Carbamate Models 193

Table 26. Biotransformation and Elimination Paths of Carbofuran and the Result-
ing Metabolites and Preliminary Liver Vmax and Km Valuesa.

No.b Initiating pesticide/metabolitec Resulting metabolite

1 Carbofuran
Carbofuran-7-phenol
3-OH carbofuran
Hydroxymethyl carbofuran
Carbofuran eliminated

2 Carbofuran 7-phenol
3,7-Diol
7-Phenyl sulfuric acid
7-Phenyl glucuronic acid
7-Phenol eliminated

3 3-OH carbofuran
3,7-Diol
3-Keto carbofuran
3-OH carbofuran sulfuric acid
3-OH carbofuran glucuronic acid
3-OH carbofuran eliminated

4 Hydroxymethyl carbofuran
Hydroxymethyl carbofuran sulfuric acid
Hydroxymethyl carbofuran glucuronic
acid
Hydroxymethyl carbofuran eliminated

5 Carbofuran 3,7-diol
3-Keto-7-phenol
3,7-Diol sulfuric acid
3,7-Diol glucuronic acid
3,7-Diol eliminated

6 3-Keto carbofuran
3-Keto-7-phenol
3-Keto carbofuran eliminated

7 3-Keto-7-phenol
3-Keto-7-phenyl sulfuric acid
3-Keto-7-phenyl glucuronic acid
3-Keto-7-phenol eliminated

8 7-Phenyl sulfuric acid

7-Phenyl sulfuric acid eliminated

9 7-Phenyl glucuronic acid

7-Phenyl glucuronic acid eliminated
194 J.B. Knaak et al.

Table 26. (cont.)

No.b Initiating pesticide/metabolitec Resulting metabolite

10 3-OH carbofuran sulfuric acid

3-OH carbofuran sulfuric acid
eliminated

11 3-OH carbofuran glucuronic acid

3-OH carbofuran glucuronic acid
eliminated

12 Hydroxymethyl carbofuran sulfuric acid

Hydroxymethyl carbofuran sulfuric acid
eliminated

13 Hydroxymethyl carbofuran glucuronic acid

Hydroxymethyl carbofuran glucuronic
acid eliminated

14 3,7-Diol sulfuric acid

3,7-Diol sulfuric acid eliminated

15 3,7 Diol glucuronic acid

3,7-Diol glucuronic acid eliminated

16 3-Keto-7-phenyl sulfuric acid

3-Keto-7-phenyl sulfuric acid eliminated

17 3-Keto-7-phenyl glucuronic acid

3-Keto-7-phenyl glucuronic acid
eliminated
a
Preliminary estimates of liver Vmax (μmol hr−1kg−1 bw, unscaled) and Km (μM) are equal
to 10.0.
Additional published values can be found in Table 34 (Appendix B).
b
Parent chemical or metabolite number for cross-reference in Table 16 (Appendix A).
c
Refer to Table 16 for structure based on No.
Table 27. Biotransformation and Elimination Paths of Formetanate and the Result-
ing Metabolites and Preliminary Liver Vmax and Km Valuesa.

No.b Initiating pesticide/metabolitec Resulting metabolite

1 Formetanate
Demethylformetanate
3-Formaminophenyl
N
N-methylcarbamate

2 Demethylformetanate
3-Formaminophenol
Demethylformetanate eliminated

3 3-Formaminophenyl N-methylcarbamate
3-Formaminophenol

4 3-Formaminophenol
3-Aminophenol
3-Formaminophenyl glucuronic acid
3-Formaminophenyl sulfuric acid

5 3-Aminophenol
3-Acetamidophenol
3-Aminophenyl glucuronic acid
3-Aminophenyl sulfuric acid

6 3-Acetamidophenol
3-Acetamidophenyl glucuronic acid
3-Acetamidophenyl sulfuric acid

7 3-Formaminophenyl glucuronic acid

3-Formaminophenyl glucuronic acid
eliminated

8 3-Formaminophenyl sulfuric acid

3-Formaminophenyl sulfuric acid
eliminated

9 3-Aminophenyl glucuronic acid

3-Aminophenyl glucuronic acid
eliminated

10 3-Aminophenyl sulfuric acid

3-Aminophenyl sulfuric acid eliminated

11 3-Acetamidophenyl glucuronic acid

3-Acetamidophenyl glucuronic acid
eliminated

12 3-Acetamidophenyl sulfuric acid

3-Acetamidophenyl sulfuric acid
eliminated
a
Preliminary estimates of liver Vmax (μmol hr−1kg−1 bw, unscaled) and Km (μM) are equal
to 10.0.
b
Parent chemical or metabolite number for cross-reference in Table 17 (Appendix A).
c
Refer to Table 17 for structure based on No.

195
196 J.B. Knaak et al.

Table 28. Biotransformation and Elimination Paths of Methiocarb and the Result-
ing Metabolites and Preliminary Liver Vmax and Km Valuesa.

No.b Initiating pesticide/metabolitec Resulting metabolite

1 Methiocarb
Methiocarb sulfoxide
Methiocarb phenol
Hydroxymethyl methiocarb

2 Methiocarb sulfoxide
Methiocarb sulfone
Methiocarb sulfoxide phenol

3 Methiocarb sulfone
Methiocarb sulfone phenol

4 Hydroxymethyl methiocarb
Hydroxymethyl methiocarb glucuronic
acid
Hydroxymethyl methiocarb sulfuric acid
Hydroxymethyl methiocarb eliminated

5 Methiocarb phenol
Methiocarb phenyl glucuronic acid
Methiocarb phenyl sulfuric acid
6 Methiocarb phenol sulfoxide
Methiocarb sulfoxide phenyl glucuronic
acid
Methiocarb sulfoxide phenyl sulfuric acid

7 Methiocarb phenol sulfone

Methiocarb sulfone phenyl glucuronic
acid
Methiocarb sulfone phenyl sulfuric acid

8 Hydroxymethyl methiocarb glucuronic acid

Hydroxymethyl methiocarb glucuronic
acid eliminated

9 Hydroxymethyl methiocarb sulfuric acid

Hydroxymethyl methiocarb sulfuric acid
eliminated

10 Methiocarb phenyl glucuronic acid

Methiocarb phenyl glucuronic acid
eliminated
 Parameters for Carbamate Models 197

Table 28. (cont.)

No.b Initiating pesticide/metabolitec Resulting metabolite

11 Methiocarb phenyl sulfuric acid

Methiocarb phenyl sulfuric acid
eliminated

12 Methiocarb sulfoxide phenyl glucuronic acid

Methiocarb sulfoxide phenyl glucuronic
acid eliminated

13 Methiocarb sulfoxide phenyl sulfuric acid

Methiocarb sulfoxide phenyl sulfuric acid
eliminated

14 Methiocarb sulfone phenyl glucuronic acid

Methiocarb sulfone phenyl glucuronic
acid eliminated

15 Methiocarb sulfone phenyl sulfuric acid

Methiocarb sulfone phenyl sulfuric acid
eliminated
a
Preliminary estimates of liver Vmax (μmol hr−1kg−1 bw, unscaled) and Km (μM) are equal
to 10.0.
b
Parent chemical or metabolite number for cross-reference in Table 18 (Appendix A).
c
Refer to Table 18 for structure based on No.

Table 29. Biotransformation and Elimination Paths of Methomyl and the Resulting
Metabolites and Preliminary Liver Vmax and Km Valuesa.

No.b Initiating pesticide/metabolitec Resulting metabolite

1 syn-(Z)-methomyl
anti-(E)-methomyl
syn-(Z)-methomyl oxime

2 anti-(E)-methomyl
anti-(E)-methomyl oxime

3 syn-(Z)-methomyl oxime
S-Methyl, N
N-methyl carbamic acid

4 anti-(E)-methomyl oxime
Ethylium
198 J.B. Knaak et al.

Table 29. (cont.)

No.b Initiating pesticide/metabolitec Resulting metabolite

5 S-Methyl, N
N-methyl carbamic acid
Carbon dioxide

6 Ethylium
Acetonitrile

7 Carbon dioxide
Carbon dioxide eliminated

8 Acetonitrile
Hydrogen cyanide
Glutathione conjugate of acetonitrile
Acetonitrile eliminated

9 Hydrogen cyanide
Hydrogen cyanide eliminated

10 Glutathione conjugate of acetonitrile

Glutamylcysteine conjugate of acetonitrile
Cysteinylglycine conjugate of acetonitrile
Glutathione conjugate of acetonitrile
eliminated

11 Glutamylcysteine conjugate of acetonitrile

Cysteine conjugate of acetonitrile
Glutamylcysteine conjugate of acetonitrile
eliminated

12 Cysteinylglycine conjugate of acetonitrile

Cysteine conjugate of acetonitrile
Cysteinylglycine conjugate of acetonitrile
eliminated

13 Cysteine conjugate of acetonitrile

Cysteine-sulfate conjugate of acetonitrile
Cysteine conjugate of acetonitrile
eliminated

14 N-Sulfate-cysteine conjugate of acetonitrile

N
Cysteine-sulfate conjugate of acetonitrile
eliminated
a
Preliminary estimates of liver Vmax (μmol hr−1kg−1 bw, unscaled) and Km (μM) are equal to
10.0.
b
Parent chemical or metabolite number for cross-reference in Table 19 (Appendix A).
c
Refer to Table 19 for structure based on No.
 Parameters for Carbamate Models 199

Table 30. Biotransformation and Elimination Paths of Oxamyl and the Resulting
Metabolites and Preliminary Liver Vmax and Km Valuesa.

No.b Initiating pesticide/metabolitec Resulting metabolite

1 Oxamyl
des N oxamyl
Oxamyl oxime
Oxamyl nitrile

2 des N oxamyl
des N Oxamyl oxime

3 Oxamyl oxime
Oxamyl oxime glucuronic acid
des N oxamyl oxime

4 Oxamyl nitrile, DMCF

Oxamyl acid

5 des N oxamyl oxime

des N oxamyl oxime glucuronic acid
6 Oxamyl oxime glucuronic acid
Oxamyl oxime glucuronic acid eliminated

7 Oxamyl acid (N,N-dimethyloxamic

N acid)
des N oxamyl acid
Glucuronide of Oxamyl acid

8 des N oxamyl oxime glucuronic acid

des N oxamyl oxime glucuronic acid
eliminated

9 Glucuronide of N,N-dimethyloxamic
N acid
Glucuronide of N,N-dimethyloxamic
N acid
eliminated
10 des N
N-oxamyl acid (N
N-methyloxamic acid)
Glucuronide of N
N-methyloxamic acid

11 Glucuronide of N
N-methyloxamic acid
Glucuronide of N
N-methyloxamic acid
eliminated
a
Preliminary estimates of liver Vmax (μmol hr−1kg−1 bw, unscaled) and Km (μM) are equal
to 10.0.
b
Parent chemical or metabolite number for cross-reference in Table 20 (Appendix A).
c
Refer to Table 20 for structure based on No.
200 J.B. Knaak et al.

Table 31. Biotransformation and Elimination Paths of Pirimicarb and the Resulting
Metabolites and Preliminary Liver Vmax and Km Valuesa.

No.b Initiating pesticide/metabolitec Resulting metabolite

1 Pirimicarb
Pirimicarb phenol
Hydroxymethyl pirimicarb

2 Hydroxymethyl pirimicarb
Pirimicarb phenol
Demethyl pirimicarb phenol
Amino pirimicarb phenol

3 Pirimicarb phenol
Demethyl pirimicarb phenol
Hydroxymethyl pirimicarb phenol
Pirimicarb phenyl glucuronic acid
Pirimicarb phenol eliminated

4 Demethyl pirimicarb phenol

Amino pirimicarb phenol
Hydroxymethyl demethyl pirimicarb
phenol
Demethyl pirimicarb phenol eliminated

5 Amino pirimicarb phenol

Amino pirimicarb phenol eliminated

6 Hydroxymethyl demethyl pirimicarb phenol

Hydroxymethyl demethyl pirimicarb
phenol eliminated

7 Pirimicarb phenyl glucuronic acid

Pirimicarb phenyl glucuronic acid
eliminated

8 Hydroxymethyl pirimicarb phenol

Hydroxymethyl pirimicarb phenol
eliminated
a
Preliminary estimates of liver Vmax (μmol hr−1kg−1 bw, unscaled) and Km (μM) are equal
to 10.0.
b
Parent chemical or metabolite number for cross-reference in Table 21 (Appendix A).
c
Refer to Table 21 for structure based on No.
Table 32. Biotransformation and Elimination Paths of Propoxur and the Resulting
Metabolites and Preliminary Liver Vmax and Km Valuesa.

No.b Initiating pesticide/metabolitec Resulting metabolite

1 Propoxur
2-Isopropoxy phenol
5-OH propoxur
4-OH propoxur
Hydroxymethyl propoxur
Propoxur eliminated

2 2-Isopropoxy phenol
2-Isopropoxy phenyl sulfuric acid
2-Isopropoxy phenyl glucuronic acid
2-Isopropoxy phenol eliminated

3 2-OH phenyl methylcarbamate

2-OH phenyl methylcarbamate sulfuric
acid
2-OH phenyl methylcarbamate
glucuronic acid
2-OH phenyl methylcarbamate
eliminated

4 5-OH propoxur
2-Isopropoxy 5-OH phenol
5-OH propoxur sulfuric acid
5-OH propoxur glucuronic acid
5-OH propoxur eliminated

5 2-Isopropoxy 5-OH phenol

2-Isopropoxy 5-OH phenyl sulfuric acid
2-Isopropoxy 5-OH phenyl glucuronic
acid
2-Isopropoxy 5-OH phenol eliminated

6 4-OH propoxur
2-Isopropoxy 4-OH phenol
4-OH propoxur sulfuric acid
4-OH propoxur glucuronic acid
4-OH propoxur eliminated

7 2-Isopropoxy 4-OH phenol

2-Isopropoxy phenyl sulfuric acid
2-Isopropoxy phenyl glucuronic acid
2-Isopropoxy 4-OH phenol eliminated

8 N-Hydroxymethyl propoxur
Hydroxymethyl propoxur sulfuric acid
Hydroxymethyl propoxur glucuronic acid
Hydroxymethyl propoxur eliminated

201
202 J.B. Knaak et al.

Table 32. (cont.)

No.b Initiating pesticide/metabolitec Resulting metabolite

9 2-Isopropoxy phenyl sulfuric acid

2-Isopropoxy phenyl sulfuric acid
eliminated

10 2-Isopropoxy phenyl glucuronic acid

2-Isopropoxy phenyl glucuronic acid
eliminated

11 2-OH phenyl methylcarbamate sulfuric acid

2-OH phenyl methylcarbamate sulfuric
acid eliminated

12 2-OH phenyl methylcarbamate glucuronic acid

2-OH phenyl methylcarbamate
glucuronic acid eliminated

13 5-OH propoxur sulfuric acid

5-OH propoxur sulfuric acid eliminated

14 5-OH propoxur glucuronic acid

5-OH propoxur glucuronic acid
eliminated

15 2-Isopropoxy 5-OH phenyl sulfuric acid

2-Isopropoxy 5-OH phenyl sulfuric acid
eliminated

16 2-Isopropoxy 5-OH phenyl glucuronic acid

2-Isopropoxy 5-OH phenyl glucuronic
acid eliminated

17 4-OH propoxur sulfuric acid

4-OH propoxur sulfuric acid eliminated

18 4-OH propoxur glucuronic acid

4-OH propoxur glucuronic acid
eliminated

19 2-Isopropoxy 4-OH phenyl sulfuric acid

2-Isopropoxy 4-OH phenyl sulfuric acid
eliminated
 Parameters for Carbamate Models 203

Table 32. (cont.)

No.b Initiating pesticide/metabolitec Resulting metabolite

20 2-Isopropoxy 4-OH phenyl glucuronic acid

2-Isopropoxy 4-OH phenyl glucuronic
acid eliminated
21 N-Hydroxymethyl propoxur sulfuric acid
N
Hydroxymethyl propoxur sulfuric acid
eliminated
22 N-Hydroxymethyl propoxur glucuronic acid
N
Hydroxymethyl propoxur glucuronic acid
eliminated
a
Preliminary estimates of liver Vmax (μmol hr−1kg−1 bw, unscaled) and Km (μM) are equal
to 10.0.
b
Parent chemical or metabolite number for cross-reference in Table 22 (Appendix A).
c
Refer to Table 22 for structure based on No.

Table 33. Biotransformation and Elimination Paths of Thiodicarb and the Result-
ing Metabolites and Preliminary Liver Vmax and Km Valuesa.

No.b Initiating pesticide/metabolitec Resulting metabolite

1 Thiodicarb
syn-(Z)-methomyl
anti-(E)-methomyl

2 syn-(Z)-methomyl
ani-(E)-methomyl
syn-(Z)-methomyl oxime

3 anti-(E)-methomyl
anti-(E)-methomyl oxime

4 syn-(Z)-methomyl oxime
S-Methyl, N
N-methyl carbamic acid

5 anti-(E)-methomyl oxime
Ethylium

6 S-Methyl, N
N-methyl carbamic acid
Carbon dioxide

7 Ethylium
Acetonitrile
204 J.B. Knaak et al.

Table 33. (cont.)

No.b Initiating pesticide/metabolitec Resulting metabolite

8 Carbon dioxide
Carbon dioxide eliminated

9 Acetonitrile
Hydrogen cyanide
Glutathione conjugate of acetonitrile
Acetonitrile eliminated

10 Hydrogen cyanide
Hydrogen cyanide eliminated

11 Glutathione conjugate of acetonitrile

Glutamylcysteine conjugate of acetonitrile
Cysteinylglycine conjugate of acetonitrile
Glutathione conjugate of acetonitrile
eliminated

12 Glutamylcysteine conjugate of acetonitrile

Cysteine conjugate of acetonitrile
Glutamylcysteine conjugate of acetonitrile
eliminated

13 Cysteinylglycine conjugate of acetonitrile

Cysteine conjugate of acetonitrile
Cysteinylglycine conjugate of acetonitrile
eliminated

14 Cysteine conjugate of acetonitrile

Cysteine-sulfate conjugate of acetonitrile
Cysteine-sulfate conjugate of acetonitrile
eliminated

15 N-Sulfate-cysteine conjugate of acetonitrile

Cysteine-sulfate conjugate of acetonitrile
eliminated
a
Preliminary estimates of liver Vmax (μmol hr−1kg−1 bw, unscaled) and Km (μM) are equal
to 10.0.
b
Parent chemical or metabolite number for cross-reference in Table 23 (Appendix A).
c
Refer to Table 23 for structure based on No.
Table 34. Published Liver Microsomal Vmax and Km Values for Carbaryl and Carbofuran.

Biotransformation and elimination paths Liver microsomes

Initiating pesticide/ Vmaxb nmol min−1 mg−1

Parent chemical metabolite (no.a) Resulting metabolite protein Km μmol L−1 Source

Carbaryl Carbaryl, HLMc 4-OH Carbaryl 0.87 349 Tang et al. 2002
Carbaryl, HLM 5-OH Carbaryl 0.04 349 Tang et al. 2002
Carbaryl, HLM Hydroxymethyl carbaryl 0.57 81.0 Tang et al. 2002
Carbofuran Carbofuran (rat output)d Carbofuran-7-phenol 0.012 25.0 Zhang et al. 2006
Carbofuran (rat output)d 3-OH carbofuran 0.024 12.5 Zhang et al. 2006
Carbofuran, HLM 3-OH carbofuran 3.30 1974 Usmani et al. 2004
Carbofuran, RLMc 3-OH carbofuran 5.50 207 Usmani et al. 2004
a
From metabolism pathways in Tables 25 and 26.
b
Parameters for Carbamate Models

HLM Vmax values (nmol min−1 mg−1 protein) and may be expressed as in vivo values (μmol hr−1 kg−1 bw); this is accomplished by multiplying the in vitro
values by 60 min hr−1; 30 mg microsomal protein/g liver; and by 27 g liver/kg bw to give the in vivo value in nmol hr−1kg−1 bw and dividing the resultant by
1000.
c
HLM, human liver microsomes; RLM, rat liver microsomes.
d
ERDEM Vmax (mmol/L/H) was converted to nmol min−1 mg−1 protein by using 30 mg protein/g liver.
205
 206

Table 35. Published CYP Vmax and Km Values for Carbaryl and Carbofuran.

Biotransformation and elimination paths Liver

Initiating pesticide Vmaxb nmol min−1 nmol−1

Parent and CYP (no.a) Resulting metabolite isoform Km μmol L−1 Source

Carbaryl Carbaryl, CYP1A1 4-OH carbaryl 3.89 20 Tang et al. 2002

Carbaryl, CYP1A2 1.72 58
Carbaryl, CYP2B6 0.80 11
Carbaryl, CYP2C19 2.21 44
Carbaryl, CYP3A4 5.81 235
Carbaryl, CYP1A1 5-OH carbaryl 4.81 15 Tang et al. 2002
Carbaryl, CYP1A2 2.62 89
Carbaryl, CYP2B6 0.29 110
Carbaryl, CYP2C19 0.99 62
Carbaryl, CYP3A4 2.34 281
J.B. Knaak et al.

Carbaryl, CYP1A1 Hydroxymethyl carbaryl 1.19 51 Tang et al. 2002

Carbaryl, CYP1A2 4.80 36
Carbaryl, CYP2B6 15.54 45
Carbaryl, CYP2C19 3.46 15
Carbaryl, CYP3A4 1.47 156
Carbofuran Carbofuran, CYP3A4 3-OH carbofuran 40.3 742 Usmani et al. 2004
Carbofuran, CYP1A2 3-OH carbofuran 22.0 238 Usmani et al. 2004
Carbofuran, CYP2C19 3-OH carbofuran 18.0 199 Usmani et al. 2004
a
From metabolism pathways in Tables 25 and 26. bThese values may be converted to in vivo values (nmol hr−1 kg−1 bw) by multiplying the values by
CYP content in nmol mg−1 microsomal protein, 60 min hr−1; 30 mg microsomal protein/g liver; and by 27 g liver/kg bw to give the in vivo value.
 Parameters for Carbamate Models 207

Appendix C. Physiological model for aldicarb depicting its

metabolic pathway.

Fig. 9. Physiologically based pharmacokinetic/pharmacodynamic model for oral and dermal exposure to
aldicarb. The model includes the inhibition and recovery of blood AChE and BChE, brain AChE, BChE,
and CaE, and liver BChE and CaE.
208 J.B. Knaak et al.

Appendix D. Nomenclature.

i. Acronyms and Abbreviations:

Term Description

ACh acetylcholine (CAS no. 51-84-3)

AChE acetylcholinesterase
ACS active site-selective aromatic cation-binding site
ACSL Advanced Continuous Simulation Language
ADHP 2-amino-5,6-dimethyl-4-hydroxypyrimidine
(CAS no. 78195-313-0)
ADME absorption, distribution, metabolism, elimination
AFP adaptive fuzzy partitioning
AMET absorption, metabolism, elimination and toxicity
AntiChE anticholinesterase
AOH acetic acid
AS anionic substrate-binding site
ATCh acetylthiocholine (CAS no. 4468-05-7)
BCh butyrylcholine (CAS no. 3922-86-9)
BChE butyrylcholinesterase
BCS Biopharmaceutical Classifi fication System
BW body weight
CaE carboxylesterase
CDFA California Department of Food and Agriculture
CE See CaE
CES See CaE
ChE cholinesterase
CNS central nervous system
CYP cytochrome P450
CYP450 cytochrome P450
DDHP 2-dimethylamino-5,6-dimethyl-4-hydroxypyrimidine
(CAS no. 78195-32-1)
DEAE diethylaminoethyl
DFP diisopropyl phosphofl fluoridate (CAS no. 55-91-4)
DHHP 2-dimethylamino-6-hydroxymethyl-5-methyl-4-hydroxypyrimidine
EE electric eel
ELISA Enzyme-linked immunosorbent assay
EPA Environmental Protection Agency
ERDEM Exposure-Related Dose Estimating Model
(see web site: http://www.epa.gov/heasdweb/erdem/erdem.htm)
ES esteratic site
FAO Food and Agriculture Organization, WHO
FIFRA Federal Insecticide, Fungicide and Rodenticide Act
FMO flavin monooxygenase systems
FQPA Food Quality Protection Act
GA/SW genetic algorithm concepts and a stepwise technique
 Parameters for Carbamate Models 209

i. Acronyms and Abbreviations: (cont.)

Term Description

HB hydrogen bonding
hCE-1, hCE-2 human liver isozymes
HLM human liver microsome
HPLC high performance liquid chromatography
HuAChE human AChE
IP intraperitoneally
IPP 2-isopropoxyphenol (CAS no. 4812-20-8)
iso-OMPA tetraisopropylpyrophosphoramide (CAS no. 513-00-8)
LD50 lethal dose for 50% of the population
LOAEL lowest observed adverse effect level
MDHP 2-methylamino-5,6-dimethyl-4-hydroxypyrimidine
(CAS no. 78195-33-2)
MEPQ 7-(methylethoxyphosphinyloxy)-1-methylquinolinium iodide
(CAS no. 95230-44-7)
MLM mouse liver microsome
MR molar refractivity
MRP2 multidrug-resistant protein
NADPH nicotinomide adenine dinucleotide phosphate
NCEA National Center for Environmental Assessment
NOAEL no observable adverse effect level
OATP organic anion-transporting peptides
OP organophosphorus
ORMUCS ordered multicategorical classifi
fication method using the simplex
technique
PAPS 3′-phosphoadenosinne-5′-phosphosulfate
PAS peripheral anionic binding site or sites
PBPK/PD physiologically based pharmacokinetic/pharmacodynamic
PCB polychlorinated biphenyl
QSAR quantitative structure–analysis relationship
RBC red blood cells
RfD reference dose
RLM rat liver microsome
RPF relative potency factor
SDS-PAGE sodium dodecyl sulfate-polyacrylamide gel electrophoresis
TCh thiocholine
TLC thin-layer chromatography
UDP umbelliferyl diethyl phosphate (CAS no. 299-49-6)
UDPGA uridine diphosphate glucuronic acid
UGT UDP-glucuronosyltransferases
WHO World Health Organization
210 J.B. Knaak et al.

ii. Chemical and Mathematical Expressions:

Expression Description

α molecular polarizability
[E] enzyme concentration
[E]t; [E]T total enzyme
[EH], EH concentration of the active enzyme
[ES] enzyme–substrate complex
[IB], IB concentration of carbamate in the incubation mixture
[S] substrate concentration
ACt total AChE
AiACHEB amount of inhibited blood AChE
Arsal amount of chemical in reference saline
Assal amount of chemical in the sample saline after fifiltration
b coeffificient from nonlinear equation fitting Eq. 32; reflflects
effi
ficiency of hydrolysis of ternary complex SES relative to ES
BH leaving group
CB carbamic acid concentration or concentration of pesticide in skin
CACHEB concentration of free AChE in blood
Camax maximum H-bond acceptor descriptor in a molecule
CB carbamate inhibitor
CCarbamate concentration of carbamate in blood
Cfil
fi concentration of the chemical in the ultrafi
filtrate from reference
saline
CHG indicator variable for charged substituents
Cj concentration of chemical in tissue j
Csal concentration of the chemical in the reference saline before
ultrafifiltration
Css concentration of chemical in the sample saline after fifiltration
D*vo:w vegetable oil:buffer, nonionized and ionized species at pH 7.4; or
distribution between vegetable oil and buffer at pH 7.4 (Eq. 15)
Dvo:w distribution between vegetable oil:buffer, neutral compound
Dvo:w, pH7.4 distribution between n-octanol and buffer at pH 7.4 (ACD values)
E enzyme
E0 initial enzyme
EA acylated enzyme
EC carbamylated enzyme
EHAB enzyme inhibitor or Michaelis–Menten complex
EHCB enzyme–carbamate complex
EHt EH at time t
EI enzyme inhibitor complex
Es Taft steric parameter
fub binding to protein in blood
fut binding to protein in tissue
HB leaving group
ᑣ inductive parameter
I0 initial inhibitor
I50 molar concentrations resulting in 50% inhibition
 Parameters for Carbamate Models 211

ii. Chemical and Mathematical Expressions: (cont.)

Expression Description

K Km, substrate concentration that results in half-maximum velocity

k+1 equilibrium reaction, forward velocity
k−1 equilibrium reaction, backward velocity
k+2 carbamylation reaction, same as k2
k+3 decarbamylation rate
k2 carbamylation rate
Kss substrate inhibition constant
Ka binding affifinity constant
kcat hydrolysis rate constant, hr−1
Kd equilibrium or affifinity constant
ki bimolecular inhibtion rate constant
KiACHEB AChE bimolecular inhibition rate constant
Km substrate concentration that results in half-maximum velocity
Kow octanol:water partition coefficient
fi
kp turnover rate
LD50 lethal dose for 50% of the population
Log D distribution coeffificient for partially dissociated compounds
Log DpH7.4 Log D calculated with pH of 7.4
Log P log of octanol:water partition coeffi ficient, neutral compounds
nl neutral lipids
p plasma
P partition coefficient
fi
PCb:adipose blood:adipose tissue partition coefficient
fi
PCb:liver blood:liver partition coefficient
fi
PCt tissue:blood partition coefficient
fi
ph phospholipds
pI plasma
Pj partition coeffificient for tissue j
pKa acid–base ionization constant
Po:w n-octanol:buffer partition coeffi ficient, nonionized species at pH 7.4
Pt:b tissue:blood partition coefficient
fi
Pt:p adipose partition coeffificient between adipose tissue and plasma
Pt:p nonadipose partition coeffificient between nonadipose tissue and plasma
q+max maximum positive charge
Rf recovered fraction
RGMR MR of certain parts of ring substituents
ΣCa sum of H-bond factor values for all acceptor substructures in a
molecule
ΣCd sum of H-bond factor values for all donor atoms in a molecule
SES termary enzyme complex
ΣQ+ sum of all positive partial atomic charges for all atoms in the
molecule
t tissue
t inhibition time
t1/2 half-life
212 J.B. Knaak et al.

ii. Chemical and Mathematical Expressions: (cont.)

Expression Description

V fractional tissue volume content

VB volume of blood
Vi initial velocity
Vj volume of tissue j
Vmax half-maximum velocity
Vnlb fractional volume of neutral nonpolar lipids in blood
Vnlt fractional volume of neutral nonpolar lipids in tissue
Vphlb fractional volume of phospholipids in blood
Vphlt fractional volume of phospholipids in tissue
Vss volume of saline in the test vial
Vwb fractional volume of water in blood
Vwt fractional volume of water in tissue
w water
WS water solubility
Rev Environ Contam Toxicol 193:213–285 © Springer 2008

Persistent Organic Pollutants in Vietnam:

Environmental Contamination and
Human Exposure

Tu Binh Minh, Hisato Iwata, Shin Takahashi, Pham Hung Viet,

Bui Cach Tuyen, and Shinsuke Tanabe

Contents
I. Introduction ...................................................................................................... 213
II. Production and Use ......................................................................................... 215
III. Contamination Status ...................................................................................... 216
A. Air, Water, Sediments, and Soils ............................................................ 216
B. Biological Samples .................................................................................... 230
C. Foodstuffs ................................................................................................... 242
D. Human Exposure ....................................................................................... 243
E. Dioxin Contamination .............................................................................. 244
IV. Environmental Behavior and Bioaccumulation .......................................... 269
A. Transport Behavior in Tropical Environments ..................................... 269
B. Bioaccumulation in Biota ......................................................................... 272
V. Temporal Trends ............................................................................................. 273
VI. Environmental and Human Health Implications ........................................ 277
VII. Conclusions and Recommendations ............................................................. 282
Summary ........................................................................................................... 284
Acknowledgments ........................................................................................... 284
References ........................................................................................................ 285

I. Introduction
Global contamination and toxic effects of persistent organic pollutants
(POPs) have been an emerging environmental issue and have received
considerable attention during the past four decades. Although the extent
of contamination by POPs has been dominant in industrialized nations, an

Communicated by G.W. Ware.

T.B. Minh, H. Iwata, S. Takahashi, S. Tanabe ( )
Center for Marine Environmental Studies (CMES), Ehime University, Bunkyo-cho 2-5, Mat-
suyama 790-8577, Japan.
P.H. Viet
Center for Environmental Technology and Sustainable Development (CETASD), Hanoi
National University, 334 Nguyen Trai Street, Thanh Xuan, Hanoi, Vietnam.
B.C. Tuyen
Nong Lam University, Linh Trung, Thu Duc District, Hochiminh City, Vietnam.

213
214 T.B. Minh et al.

increasing number of recent investigations have highlighted the role of the

Asia-Pacifific region as a potential source of emission for these chemicals,
particularly to pristine areas such as the Arctic and the Antarctic (Tanabe
et al. 1994; Tanabe 2000, 2002; Tanabe and Subramanian 2006).
In view of environmental contamination, Vietnam has been well known
as a land of extensive spraying of Agent Orange during the Vietnam War.
The high degree of dioxin contamination in some military bases and Agent
Orange-sprayed areas in South Vietnam has received particular attention
during the past 30 years. In addition to the dioxin contamination caused by
Agent Orange, the rapid agricultural and industrial growth in this country
lends credence to the widespread contamination of POPs. Vietnam is
located at the center of the Southeast Asian region (Fig. 1); it has more
than 300 km of coastal area and two major agricultural production areas:
the Red River Delta in the north and the Mekong River Delta in the south.
These two deltas are inhabited by more than 30 million people and are two
of the most densely populated areas in the world. The Mekong River Delta
has recently become one of the most productive agricultural regions of
Southeast Asia. Such a strategic geographical position and the rapid
agricultural development of Vietnam made this country an important
region where extensive studies on environmental pollution have been
carried out during the last two decades.
This chapter provides a comprehensive review of the studies dealing with
POPs in Vietnam. Available data on POP contamination in Vietnam are
compiled on the basis of various investigations in the framework of the

Red River Delta

Pacific Ocean

China

India
South
China Philippines
Sea

Malaysia
a

sia
a
Indonesia

Mekong
River Delta

Fig. 1. Map of Vietnam. Vietnam is located at the center of the Southeast Asian
region: it has two the largest deltas, Red River Delta and Mekong River Delta.
 Organic Pollutants in Vietnam 215

Asia-Pacifific Mussels Watch Program, the 21st Century Center of Excellence

Program, and the Core University Program supported by the Japan Society
for the Promotion of Science (JSPS), which were conducted in our
laboratory during the past decade. Results of these comprehensive studies
are reviewed, and various issues of POPs contamination in Vietnam are
discussed in a comparative point of view with the other countries in
the Asia-Pacifi fic region. In addition, results from other laboratories are
also reviewed to help improve insights into the distribution, transport,
bioaccumulation, and possible toxic implications on environmental quality
and human health. This review focuses on the organochlorine insecticides
such as 1,1,1-trichloro-2,2-bis(p( -chlorophenyl)ethane (DDT) and its
metabolites (DDTs), hexachlorocyclohexane isomers (HCHs), chlordane
compounds (CHLs), and hexachlorobenzene (HCB). Residue levels of
industrially derived contaminants such as polychlorinated biphenyls (PCBs),
polychlorinated dibenzo-p - -dioxins and dibenzofurans (PCDD/Fs), and
polybrominated biphenyl ethers (PBDEs) are also reviewed.

II. Production and Use

In general, information on the production and usage of POPs, particularly
organochlorine (OC) insecticides and PCBs in Vietnam, as well as some
other developing countries in the East and South Asian region, is still
limited or obscure. Systematic inventory of toxic manmade chemicals is
lacking in these countries because of their limited capacity to conduct com-
prehensive monitoring surveys. Recently, the United Nations Environment
Program (UNEP) has initiated various monitoring programs for POPs at
regional and global levels, and the results have been summarized at different
workshops. According to these reports, the active ingredients for insecticides
were not produced in Vietnam. In fact, before 1985, pesticides such as DDT
and HCB were imported from the former Soviet Union and some socialistic
countries up to a quantity of 6,500–9,000 t/yr (Sinh et al. 1999). The statistical
data showed that the total quantity of DDT imported into Vietnam for
malaria control from 1957 to 1990 was 24,042 t. During 1986–1990,
approximately 800 t was used (Sinh et al. 1999). These amounts are still
lower than those in some other countries in regions such as Malaysia,
Indonesia, and India. DDT usage for malaria control ceased in Vietnam in
1995, and other insecticides such as pyrethroid compounds have been used
as substitutes for DDT (Sinh et al. 1999).
The information on PCB usage in Vietnam is still obscure. Data indicate
that about 27,000–30,000 t of oils contaminated by PCBs were imported
from the former USSR, China, and Rumania (Sinh et al. 1999). In addition,
electrical equipment containing PCBs, such as transformers, was also
imported from Australia until the mid-1980s (Kannan et al. 1995). Yet
another possible source of PCBs in Vietnam are the weapons used
extensively during the Indochina War (Thao et al. 1993a,b). The major
216 T.B. Minh et al.

source of dioxins in Vietnam in the past was Agent Orange and other
herbicides sprayed in South Vietnam during the American war. Recently,
Stellman and coworkers (Stellman et al. 2003) provided revised estimates
of the amounts of herbicides used in Vietnam. During 1961–1971, at least
about 45 million L Agent Orange was sprayed (Stellman et al. 2003). 2,4,5-
T, a constituent of Agent Orange, is known to be contaminated with 2,3,7,8-
tetrachlorodibenzo-p- -dioxin (TCDD). However, the combustion-derived
sources of dioxins in Vietnam are unknown. Various kinds of combustion
processes may facilitate the widespread contamination of dioxins and
related compounds in Vietnam.

III. Contamination Status

A. Air, Water, Sediments, and Soils
Comprehensive monitoring surveys have been conducted to examine the
distribution of POPs such as PCBs, DDTs, HCHs, and HCB in air, water,
and sediments from estuarine environments from various countries in
the Asia-Pacifi fic comprising Japan, India, Vietnam, Thailand, Indonesia,
Malaysia, the Philippines, and Australia (Iwata et al. 1994). These
investigations reported the presence of higher residues of DDTs and HCHs
in air and water from coastal and estuarine areas in the developing countries
of tropical and subtropical regions (India, Thailand, and Vietnam), than in
developed nations (Japan and Australia). A compilation of available data
for Vietnam is given in Tables 1 and 2. The distribution in air, water, and
sediments from north, middle, and south regions of Vietnam showed
relatively higher DDT concentrations, supporting the concept of widespread
contamination of this insecticide throughout the country. This result suggests
extensive use of DDT for agricultural purposes in the past and for malaria
control until very recently. Interestingly, in a survey conducted about 10 yr
later than the survey by Iwata et al. (1994) (in 1998/1999) covering an
extended area along Red River and Duong River, the two biggest rivers in
northern Vietnam, elevated concentrations of DDTs, HCHs, and CHLs
were reported (Hung and Thiemann 2002). The levels of DDTs, HCHs,
and CHLs in Red River and Duong River were apparently higher than
those reported in the early 1990s surveys. In addition, wastewater collected
from extensive human activity areas such as canals of Tu Liem district, a
suburb of Hanoi city (northern Vietnam), and Thi Nghe River, Hochiminh
(southern Vietnam) contained elevated concentrations of DDTs (see Table
1). It is also interesting to note that levels found in a recent survey (in
suburb Hanoi; Hung and Thiemann 2002) were higher than those reported
a decade ago (Iwata et al. 1994). Although backgrounds of analytical
methods and sampling locations are different among studies, these
observations suggest that the use of DDT for malaria control was relatively
extensive until very recently in both northern and southern Vietnam.
Table 1. Concentrations (ng L−1) of Persistent Organochlorines in Water from Vietnam.

Location Sample Year n DDTs HCHs CHLs PCBs Reference Remarks

Red River, Hanoi River water 1990 1 0.68 3.2 0.045 0.84 lwata et al. 1994 DDTs: sum of p,p′-DDE, p,p′-DDD
Phu Loc Lake, Hue Lake water 1990 1 0.29 18 0.21 1.2 lwata et al. 1994 and p,p′-DDT
Huong River, Hue River water 1900 1 1.1 1.9 0.07 1.6 lwata et al. 1994 HCHs: sum of α-, β-, and γ-HCH
γ
Nha Be River, Hochiminh Estuarine 1990 1 4.7 31 0.55 2.7 lwata et al. 1994 CHLs: sum of cis-chlordane, trans-
City water chlordane, cis-nonachlor, trans-
Gua canal, Cu Chi River water 1990 1 0.6 9.5 0.16 1.9 lwata et al. 1994 nonachlor and oxychlordane
Long Tau River, Estuarine 1990 1 0.55 5.2 0.13 0.57 lwata et al. 1994 PCBs: quantifified by an equivalent
Hochiminh City water mixture of Kanechlor preparations
Thi Nghe canal, Hochiminh Estuarine 1990 1 25 19 1 8 lwata et al. 1994 (KC-300, KC-400, KC-500 and
City water KC-600)
Red River and Duong River, River water 1998/99 Hung and DDTs: sum of p,p′-DDE, p,p′-DDD,
northern Vietnam Thiemann 2002 p,p′-DDT, o,p′-DDE, o,p′-DDD,
a
Dry season 18 44 (0.55–320) 17 (1.6–83) 21 (0.81–110) — o,p′-DDT
Rainy season 18 56 (8.4–230) 29 (3.1–97) 27 (1.6–130) — HCHs: sum of α-, β-, γ-
γ , and δ-HCH
CHLs: sum of heptachlor and
heptachlorepoxide
Lakes in Hanoi: West Lake, Lake water 1998/99 Hung and
Thuyen Quang Lak Thiemann 2002
Bay Mau Lake and Ba Mau
Lake
Organic Pollutants in Vietnam

Dry season 4 2.1 (0.21–3.6) 0.69 (0.26–2.16) 2.9 (1.1–4.8) —

Rainy season 4 5.1 (0.65–15) 32 (0.69–120) 13 (2–51) —
Irrigation canal, Tu Liem, River water 1998/99 Hung and
suburb Hanoi Thiemann 2002
Dry season 6 59 (8.2–130) 7.2 (1.6–1.8) 6.9 (6.0–7.5) —
Rainy season 6 50 (11–110) 17 (5.5–26) 15 (1.2–25) —
Wells, Gia Lam, suburb Groundwater 1998–1999 Hung and
Hanoi Thiemann 2002
Dry season 2 0.17 (0.11–0.23) 0.21 (0.19–0.22) 0.17 —
Rainy season 2 0.09 0.04 <0.5 —

a
217

Mean (range).
218 T.B. Minh et al.

Table 2. Concentrations (ng g−1 dry wt) of Persistent Organochlorines in Sediments

and Soils From Vietnam.

Sample/sampling site
Location characteristic Year n DDTs

Phu Da, Hue Sediment, paddy field 1990 1 0.52

A Luoi, Binh Tri Thien Sediment, municipal 1990 1 68

sewage
Long Giang Canal, Duyen Sediment, paddy fi
field 1990 1 8
Hai
Lo Giang River, Duyen Hai Sediment, paddy field
fi 1990 1 1

Don Canal, Duyen Hai Sediment, paddy field

fi 1990 1 3.3

La Canal, Duyen Hai Sediment, paddy field

fi 1990 1 10

Ben Nghe Canal, Hochiminh Sediment, municipal 1990 1 120

City sewage
Tan Binh, Tay Ninh Sediment, paddy fi
field 1990 1 7.8

Gua Canal, Cu Chi Sediment, paddy fi

field 1990 1 0.37

Song Long Tau, Duyen Hai, Sediment, paddy field 1990 6 17 (2.1–47)
Hochiminh City
Thi Nghe Canal, Hochiminh Sediment, municipal 1990 3 530 (360–790)
City sewage
Thinh Liet, Thanh Tri, Hanoi Soil, paddy fi
field 1990 1 330

Vinh Quynh, Thanh Tri, Soil, paddy fi

field 1990 2 76 (31–120)
Hanoi
Yen Duyen, Thanh Tri, Soil, paddy fi
field 1990 2 47 (19–74)
Hanoi
Phu Da, Binh Tri, Thien Soil, paddy fi
field 1990 1 38

Hong Ha, A luoi, Binh Tri, Soil, paddy fi

field 1990 1 23
Thien
Ta Bat, Aluoi, Binh Tri, Soil, paddy fi
field 1990 1 11
Thien
Son Thuy, Aluoi, Binh Tri, Soil, paddy fi
field 1990 1 1300
Thien
Hong Kim, Aluoi, Binh Tri, Soil, paddy fi
field 1990 1 9.7
Thien
An Hoa, Hue Soil, paddy fi
field 1990 1 69

Phu Loc, Hue Soil, upland 1990 4 1.9 (0.73–4.4)

Binh Khanh, Duyen Hai, Soil, paddy fi

field 1990 2 27 (24–30)
Hochiminh City
Tan Thuan, Nha Be, Soil, paddy fi
field 1990 1 64
Hochiminh City
My Hung, Cu Chi Soil, paddy fi
field 1990 1 7.9
 Organic Pollutants in Vietnam 219

HCHs CHLs PCBs Reference Remarks

0.43 0.072 0.65 Iwata et al. DDTs: sum of p,p′-DDE,

1994 p,p′-DDD and
2.4 0.79 0.18 Iwata et al. p,p′-DDT
1994 HCHs: sum of α-, β-, and
1.1 0.42 3.7 Iwata et al. γ
γ-HCH
1994 CHLs: sum of cis-
0.45 0.15 2.2 Iwata et al. chlordane, trans-
1994 chlordane,
0.94 0.21 2.1 Iwata et al. cis-nonachlor, trans-
1994 nonachlor and
2.3 0.58 9.7 Iwata et al. oxychlordane
1994 PCBs: quantified
fi by an
5.2 8.8 140 Iwata et al. equivalent mixture of
1994 Kanechlor preparations
0.7 0.31 — Iwata et al. (KC-300, KC-400, KC-
1994 500 and KC-600)
0.63 0.24 0.22 Iwata et al.
1994
0.82 (0.5–1.3) 0.26 (0.14–0.46) 6.2 (2.3–8.9) Iwata et al.
1994
8.5 (6.0–12) 20 (19–20) 570 (440–630) Iwata et al.
1994
18 — 39 Thao et al. DDTs: sum of p,p′-DDE,
1993 p,p′-DDD, p,p′-DDT,
30 (4–55) — 11 (5.5–17) Thao et al. o,p′-DDT
1993 HCHs: sum of α-, β-, γ-
γ
1.8 (1.3–2.3) — 21 (13–28) Thao et al. and δ-HCH
1993 CHLs: sum of cis-
2.8 — 8.6 Thao et al. chlordane, trans-
1993 chlordane,
4.7 — 5.2 Thao et al. cis-nonachlor, trans-
1993 nonachlor and
1.4 — 2.6 Thao et al. oxychlordane
1993 PCBs: quantified
fi by an
5.7 — 12 Thao et al. equivalent mixture of
1993 Kanechlor preparations
0.31 — 0.61 Thao et al. (KC-300, KC-400, KC-
1993 500 and KC-600)
5.3 — 5 Thao et al.
1993
0.15 (0.09–0.21) — 2.1 (1.4–4.0) Thao et al.
1993
1.5 (1.4–1.5) — 3.9 (1.6–6.1) Thao et al.
1993
4 — 12 Thao et al.
1993
0.23 — 1.5 Thao et al.
1993
220 T.B. Minh et al.

Table 2. (cont.)

Sample/sampling site
Location characteristic Year n DDTs
Ba Diep, Hooc Mon Soil, upland 1990 1 280

An Phu, Cu Chi Soil, upland 1990 3 14 (1.9–37)

Tan Binh, Tan Bien, Tay Soil, paddy fi

field 1990 1 5.5
Ninh
Thanh Phu, Tan Bien, Tay Soil, paddy fi
field 1990 1 350
Ninh
Duong Minh Chau, Tay Ninh Soil, paddy fi
field 1991 1 1.9

Duong Minh Chau, Tay Ninh Soil, green bean field

fi 1991 1 2.4

Tan Bien, Tay Ninh Soil, paddy fi

field 1991 1 6.6

Hoa Thanh, Tay Ninh Soil, sugarcane and 1991 1 5.1

peanut
Tan Chau, Tay Ninh Soil, rubber 1991 1 1.6
plantation
Tan Chau, Tay Ninh Soil, sugarcane 1991 1 290

Tan Chau, Tay Ninh Soil, harvested crop 1991 5 88 (1.0–270)

Tan Bien, Tay Ninh Soil, paddy fi

field 1991 1 5.7

Go Dau, Tay Ninh Soil, harvested crop 1991 1 130

Trang Bang, Tay Ninh Soil, harvested crop 1991 1 150

Duong Minh Chau, Tay Ninh Non-cultivated soil 1991 1 1.5

Tan Bien, Tay Ninh Non-cultivated soil, 1991 1 13

Former US air base
ground
Tan Chau, Tay Ninh Non-cultivated soil, 1991 1 0.46
US bombed site
Tan Bien, Tay Ninh Non-cultivated soil 1991 6 11 (0.25–38)

Tan Uyen, southern Vietnam Non-cultivated soil 1991 2 14 (2.0–26)

Saigon River, Hochiminh River sediment 1996 11 80 (1.8–250)

City

DS/RSa
Hanoi Sediment, irrigation 1995/96 1 6.9/13
canal, northwest
Hanoi
 Organic Pollutants in Vietnam 221

HCHs CHLs PCBs Reference Remarks

0.79 — 130 Thao et al.
1993
2.3 (1.9–2.5) — 3.7 (3.0–4.2) Thao et al.
1993
0.15 — 1.6 Thao et al.
1993
3.4 — 320 Thao et al.
1993
2 — 2.3 Thao et al.
1993
0.57 — 1.8 Thao et al.
1993
0.78 — 2.3 Thao et al.
1993
1.3 — 1.6 Thao et al.
1993
1.4 — 1 Thao et al.
1993
0.27 — 0.45 Thao et al.
1993
0.92 (0.09–2.3) — 34 (0.95–150) Thao et al.
1993
0.42 — 1 Thao et al.
1993
0.46 — 32 Thao et al.
1993
0.47 — 38 Thao et al.
1993
0.8 — 5 Thao et al.
1993
1.8 — 92 Thao et al.
1993

0.95 — 25 Thao et al.

1993
0.71 (0.09–1.9) — 4.8 (0.23–13) Thao et al.
1993
1.3 (0.44–2.1) — 8.0 (3.1–13) Thao et al.
1993
— — 220 Phuong et DDTs: sum of p,p′-DDE,
al. 1998 p,p′-DDD and
p,p′-DDT
PCBs: sum of 6 congeners
DS/RS DS/RS
0.14/0.07 — 4.1/0.98 Nhan et al. DDTs: sum of p,p′-DDE,
1998 p,p′-DDD and
p,p′-DDT
222 T.B. Minh et al.

Table 2. (cont.)

Sample/sampling site
Location characteristic Year n DDTs
Hanoi Sediment, paddy 1995/96 1 7.5/14
field, southwest
fi
Hanoi
Ba Lat estuary, Red River, Sediment 1995/96 1 7.1/3
northern Vietnam
Cua Lan estuary, Thai Binh Sediment, intertidal 1995/96 1 5.8/4.9
coast lines, northern mudflat
fl areas
Vietnam
Tra Ly estuary, Thai Binh Sediment, intertidal 1995/96 1 7.3/5.1
coast lines, northern mudflat
fl areas
Vietnam
Diem Dien estuary, Thai Sediment, intertidal 1995/96 1 6.2/4.6
Binh coast lines, northern mudflat
fl areas
Vietnam
Tra Co beach, Mong Cai, Marine sediment, 1997 1 (pooled) 10
northern coast of Vietnam intertidal mudflat
fl
areas
Mong Duong, northern coast Marine sediment, 1997 1 (pooled) 8.1
of Vietnam intertidal mudflat
fl
areas
Ha Long, northern coast of Marine sediment, 1997 1 (pooled) 7.2
Vietnam intertidal mudflat
fl
areas
Hai Phong, northern coast of Marine sediment, 1997 1 (pooled) 6.7
Vietnam intertidal mudflat
fl
areas
Ba Lat estuary, northern Marine sediment, 1997 1 (pooled) 6.3
coast of Vietnam intertidal mudflat
fl
areas
Cau Dien, Nhue River, Sediment, canal, 1997 2 43 (15–71)
suburb Hanoi densely populated
industrial area
Nhue River, suburb Hanoi Sediment, canal, rural 1997 1 8.3
area
Nhue River, suburb Hanoi Sediment, canal 1997 1 21

Hanoi downtown Sediment, canal, 1997 1 13

densely populated
industrial area
ToLich River, suburb Hanoi Sediment, canal, 1997 2 59 (36–81)
densely populated
industrial area
Thuong Tin, suburb Hanoi Sediment, canal, 1997 2 37 (24–50)
southern Hanoi
city
Thuong Tin, suburb Hanoi Sediment, canal, rural 1997 1 7.4
area, southern
Hanoi city
 Organic Pollutants in Vietnam 223

HCHs CHLs PCBs Reference Remarks

0.41/0.16 — 6.0/1.3 Nhan et al. PCBs: quantified
fi by
1998 Arochlor 1260
HCHs: γγ-HCH only
0.5/0.05 — 1.1/0.7 Nhan et al.
1998
0.48/0.025 — 0.87/0.41 Nhan et al.
1998

0.62/0.13 — 0.36/0.32 Nhan et al.

1998

0.36/0.11 — 0.23/0.11 Nhan et al.

1998

34 — 22 Nhan et al. DDTs: sum of p,p′-DDE,

1999 p,p′-DDD and
p,p′-DDT
4.1 — 0.51 Nhan et al. HCHs: sum of α-, β, and
1999 γ
γ-HCH
PCBs: sum of 13
1.8 — 11 Nhan et al. congeners
1999

1.7 — 18 Nhan et al.

1999

1.2 — 0.33 Nhan et al.

1999

0.34 (0.09–0.58) — 1.7 (0.97–2.51) Nhan et al. DDTs: sum of p,p′-DDE,

2001 p,p′-DDD, p,p′-DDT,
o,p′-DDE, o,p′-DDD,
0.78 — 0.74 Nhan et al. o,p′-DDT and DDMU
2001 PCBs: sum 13 congeners
0.32 — 18 Nhan et al. HCHs: sum of α-, β, and
2001 γ
γ-HCH
0.44 — 5.3 Nhan et al. CHLs: sum cis-chlordane,
2001 trans-chlordane &
trans-nonacl
2.0 (0.85–3.12) — 29 (24–34) Nhan et al.
2001

1.3 (0.59–1.98) — 20 (16–24) Nhan et al.

2001

0.65 — 3.1 Nhan et al.

2001
224 T.B. Minh et al.

Table 2. (cont.)

Sample/sampling site
Location characteristic Year n DDTs
Gia Lam, suburb Hanoi Sediment, canal, rural 1997 1 17
area, eastern Hanoi
city
Dong Anh, suburb Hanoi Sediment, canal, rural 1997 1 23
area, northern
Hanoi city
Ha Long Bay, northern estuary sediment 1998 — 28
Vietnam
Viet Tri, northern Vietnam Industrial areas 1998 — 5.2
Hanoi Soils, municipal 1999–01 7 19 (1.9–52)b
dumping sites
Hanoi Soils, reference areas 1999–01 3 3.2 (2.2–4.3)
relative to the
municipal dumping
sites
Hochiminh City Soils, municipal 1999–01 6 23 (1.1–83)
dumping sites
Hochiminh City Soils, reference areas 1999–01 3 3.8 (0.41–10)
relative to the
municipal dumping
sites
Canal, Can Tho, Mekong Sediment, canals in 2003–04 4 2.8 (1.8–4.3)
River, southern Vietnam Cantho city

Hau River, Mekong River Sediment, river 2003–04 7 0.96 (0.043–1.9)

delta, southern Vietnam
Canals, Hochiminh city, Sediment, canals, 2004 5 37
Saigon River, southern densely populated
Vietnam areas
Saigon River, Hochiminh Sediment, river 2004 5 6.9
City
Saigon River, downstream Sediment, 2004 9 1.2
and coastal areas downstream and
coastal area

a
Dry season/rainy season.
b
Mean (range).
 Organic Pollutants in Vietnam 225

HCHs CHLs PCBs Reference Remarks

0.46 — 1.9 Nhan et al.
2001
0.07 — 1.8 Nhan et al.
2001
6.1 — 37 Viet et al.
2000
0.68 — 2.3 Viet et al.
2000
0.83 (0.29–2.2) 0.45 (<0.01–1.4) 12 (2.2–20) Minh et al. DDTs: sum of p,p′-DDE,
2004 p,p′-DDD, and
0.14 (0.11–0.16) 0.15 (0.11–0.18) 0.73 (0.45–1.1) Minh et al. p,p′-DDT
2004 HCHs: sum of α-, β-, and
0.37 (0.21–0.87) 1.3 (0.15–2.5) 22 (4.2–40) Minh et al. γ
γ-HCH
2006a CHLs: sum of cis-
0.36 (0.32–0.43) 0.2 (0.16–0.25) 3.6 (2.5–4.4) Minh et al. chlordane, trans-
2006a chlordane,
cis-nonachlor, trans-
nonachlor and
oxychlodane
PCBs: quantifified by an
equivalent mixture of
Kanechlor preparations
(KC-300, KC-400, KC-
500 and KC-600)
0.04 (<0.02–0.11) 0.2 (0.12–0.35) 1.8 (0.12–3.7) Iwata et al. DDTs: sum of p,p′-DDE,
2004 p,p′-DDD, and
<0.02–0.044 0.06 (0.025–0.13) 0.21 (0.12–0.54) Iwata et al. p,p′-DDT
2004 HCHs: sum of α-, β-, and
γ
γ-HCH
— — 81 Iwata et al. CHLs: sum of cis-
2005 chlordane, trans-
— — 8.5 Iwata et al. chlordane,
2005 cis-nonachlor, and
— — 0.92 Iwata et al. trans-nonachlor
2005 PCBs: quantifified by an
equivalent mixture of
Kanechlor preparations
(KC-300, KC-400, KC-
500 and KC-600)
226 T.B. Minh et al.

Geographical distribution of OCs in sediments from Vietnam showed

little variability compared to air and water (Fig. 2, Table 2). Consistent
results were also observed in a wide area when considering distribution in
the Asia-Pacifi fic region (Iwata et al. 1994). Another survey also indicated
rather uniform distribution of DDTs in sediments along the coasts of
northern Vietnam, from the sites near the border of China down toward
the Red River estuary (see Table 2; Nhan et al. 1998, 1999). Shorter
residence time in the water phase and rapid volatilization of POPs from
sediments as a consequence of the high temperatures in tropical region
could be reasons for this uniform distribution (Iwata et al. 1994; Tanabe
2000). However, an interesting result was observed for HCHs, showing
relatively higher concentrations in Mong Cai, a site near the border of
China, than other locations toward the southern areas (see Fig. 2, Table 2).
China has been known as one of the top global HCH users (Li 1999), and
this may be the reason for such high concentrations in Mong Cai.
A recent study in the Sai Gon–Dong Nai River that examined residue
concentrations of POPs in sediments collected from the canals of the
metropolitan area in Hochiminh, the river, and downstream toward the
coastal areas showed an interesting geographical distribution (Fig. 3) (Minh
TB et al. 2005; Minh NH et al. 2007a). Concentrations of DDTs and PCBs
were found to be highest in sediments from the city canals and showed an
apparent decreasing trend toward the river and coastal areas. Residues in
sediments collected from downstream and coastal areas were much lower
than those in sediments from the river and city canals. This result clearly
demonstrated the role of the urban areas in Hochiminh City with extensi

g. 2. Distribution of persistent organochlorines in sediment from various loca-

tions in Vietnam (Iwata et al. 1994; Nhan et al. 1998, 1999).
 Organic Pollutants in Vietnam 227

Fig. 3. Distribution of DDTs and PCBs in sediments from Sai Gon–Dong Nai River
region, southern Vietnam (Minh TB et al. 2005; Minh NH et al. 2007a).

g. 4. Composition of DDT compounds in sediments from Sai Gon–Dong Nai

River region, southern Vietnam (Minh TB et al. 2005; Minh NH et al. 2007a).

results on compositions of DDT compounds also further substantiate our

findings (Fig. 4). Proportions of p,p′-DDT to total DDTs concentrations
fi
were found to be the highest in sediments collected from canals of
metropolitan areas, followed by those in the sediments from the river and
coastal areas. It is known that a higher proportion of p,p′-DDT indicates
more recent residues of DDTs in the environment and biota (Strandberg
et al. 1998).
This accumulation pattern is in good agreement with the results on
the spatial distribution of total DDT concentrations, showing higher
228 T.B. Minh et al.

contamination in city canals and decreasing trends toward the downstream

and coastal areas. This result therefore suggests more recent input of DDTs
to the urban aquatic environment. In fact, the use of DDT has been offi- fi
cially prohibited in Vietnam since 1995 (Sinh et al. 1999). However, con-
tinuous input of DDT and its metabolites to the environment as well as
their elevated concentrations in humans and wildlife have been recorded
throughout the country (Nhan et al. 2001; Monirith et al. 2003; Minh TB
et al. 2002; Minh NH et al. 2004a).
In another survey, the distribution of OCs in sediments from Hau River,
one of the two largest branches of Mekong River, was also examined (Minh
NH et al. 2004b, 2007b). Surface sediment samples were collected at various
sites along the Hau River, flflowing through the two urban areas, Can Tho
and Long Xuyen. A similar trend was observed here in the Mekong River,
showing higher concentrations of DDTs and PCBs in sediments from canals
near the urban areas of Can Tho and Long Xuyen, with levels decreasing
downstream (Fig. 5). In particular, DDT residues from the site near Can
Tho and Long Xuyen were about 10–20 fold higher than those in the
respective downstream sites. This result indicates that urban areas are

Fig. 5. Distribution of DDTs and PCBs in sediments from Hau River, the largest
branch of the Mekong River, southern Vietnam (Minh NH et al. 2004b, 2007b).
 Organic Pollutants in Vietnam 229

major pollution sources of POPs. Inputs of DDT residues are probably

derived from its application for hygienic purposes and malaria vector
control in urban areas rather than DDT used for agricultural purposes.
As for soil samples, an extensive survey in northern and southern
Vietnam indicated higher concentrations of DDTs from paddy field fi sites
than from upland areas (see Table 2; Thao et al. 1993a,b). This result clearly
reflects
fl the status of DDT use as an insecticide in the past in Vietnam. In
some specificfi sites in Tay Ninh, southern Vietnam, where a U.S. Army base
was located, elevated PCB levels were recorded (Thao et al. 1993a,b). A
recent survey in the open dumping sites for municipal wastes in Hanoi
and Hochiminh City, the two largest cities in Vietnam, revealed that
concentrations of DDTs and PCBs in soils collected from dumping sites
were much higher than those in paddy fields far from the dumping sites
(Fig. 6; Minh NH et al. 2006a). PCBs and OC insecticides likely originated
from continuous loadings of municipal waste containing residues of these
compounds. The open dumping sites therefore, may act as reservoir sources
of PCBs and OC insecticides.
To understand the magnitude of contamination of POPs in Vietnam,
concentrations in water and sediments were compared with those in other
countries in Asia-Pacifi fic (Fig. 7). Higher contamination of DDTs in the
Vietnamese coastal environment was noticed, again indicating its extensive
use of in Vietnam. Interestingly, elevated PCBs residues were also observed
in water and sediments from the Mekong River estuary, southern Vietnam,
and the levels were comparable to those reported in some locations in India,
Japan, and Australia (Fig. 7). High PCB contamination in Vietnam observed

Fig. 6. Residue concentrations of persistent organochlorines in soils from open

dumping sites and rural areas (control sites) in Hanoi and Hochiminh City, the two
largest cities in Vietnam (Minh NH et al. 2006a).
230 T.B. Minh et al.

g. 7. Comparison of persistent organochlorine residues in surface water and sedi-

ment from different countries in Asia-Pacific
fi (Iwata et al. 1994).

during our survey in the early 1990s could be derived from electrical
equipment imported from industrialized nations, e.g., the former Soviet
Union and Australia, and from leakage from army weapons extensively
used in the Vietnam War during 1961–1971 (Thao et al. 1993a,b).
Available data on the worldwide comparison of PCBs and OC insecticide
levels in open dumpsites are rather scarce. A comparison of PCBs and
DDTs levels in dumping sites from Vietnam with those in soils from other
countries is given in Fig. 8. In general, the levels of PCBs in dumping sites
were higher than those in background soils from various countries, includ-
ing industrialized nations, e.g., the U.S., Russia, and Italy, where heavy
contamination of PCBs has been common (Meijer et al. 2003). Our survey
indicated that DDT levels in dumping site soils were comparable to those
found in agricultural soils collected in the early 1990s from various coun-
tries such as Russia (Iwata et al. 1995), Ireland (McGrath 1995), and
Slovakia (Marta et al. 1997), and higher than in most of the urban soils
collected in many countries such as Egypt (Ahmed et al. 1998), Korea (Kim
and Smith 2001), and the rural areas of Vietnam (Minh NH et al. 2006a)
(see Fig. 8). This observation highlights the role of open dumping sites in
developing Asian countries as significant
fi pollution sources of PCBs and
DDTs.

B. Biological Samples
The suitability of using a wide range of biological samples as bioindicators
for monitoring POPs contamination was extensively discussed by Tanabe
and Subramanian (2006). Surveys examined POP residue levels in various
 Organic Pollutants in Vietnam 231

g. 8. Comparison of organochlorine residue levels in soils from different

countries.

biological samples such as fish,fi bivalves, and birds from the Asia-Pacifific
region, including Vietnam, have made clear the status of contamination,
distribution, and source allocation. An extensive study on OC contamination
in fi
fish from Asia and Oceania including Vietnam was carried out by Kannan
et al. (1995). Similar to sediment samples, a relatively uniform distribution
of OCs was observed in fish fi in various countries in the Asia-Pacifific region.
In Vietnam, residues of DDTs were relatively high in the surveys conducted
in 1990 and 1997 (Table 3). Mussels collected from coastal areas in north
and middle Vietnam contained elevated DDT concentrations (Table 3;
Monirith et al. 2003). Subsequent surveys conducted by Nhan et al. (1998,
1999) examined OC distribution in clams from different sites along the
northern coast, showing a very similar distribution in sediment samples. In
particular, residue concentrations of DDTs, HCHs, and PCBs were rela-
tively high in the sites near the border of China, and a decreasing trend was
noticed toward the southern coastlines (Table 4). At the two estuary areas,
Hai Phong harbor with extensive human and industrial activities, and Thai
Binh province, one of the high rice production areas in Vietnam, higher
concentrations were again observed. In general, the distribution and
magnitude of contamination in sediment and biota (fish fi and bivalves) was
very similar at both the local and regional scale, which can be attributed to
the enhanced volatilization of semivolatile organic compounds because of
the high temperatures prevailing in tropical ecosystems.
Polybrominated diphenyl ethers (PBDEs) are emerging contaminants
that have received considerable attention in recent years. We conducted a
232 T.B. Minh et al.

Table 3. Concentrations (ng g−1 lipid wt) of Persistent Organochlorines in Biological

Samples from Vietnam.

Sample/sampling site Lipid

Location description Year n content (%) DDTs

Hanoi Fish 1990 7 1.9 1,900 (680–

4,000)a
Phu Da, Hue Fish 1990 6 1.9 1,100 (210–
2,700)
Hochiminh City Fish 1990 6 1.9 1,100 (89–
4,100)

Con Lu island, Red Fish (mugil sp. and 1997 10 3.2 4,200 (330–
River estuary, chlorophthalmus 8,500)
northern sp.)
Vietnam
Con Lu island, Red Shrimp 1997 20 (pooled as 2.8 160
River estuary 1 sample)
Cat Ba island, Cai Green Mussel, 1997 38 (pooled as 1.1 530
Hai province floating habitat 1 sample)
Cat Hai province Green Mussel, 1997 34 (pooled as 0.9 300
floating 1 sample)
habitat
Cat Hai province Green Mussel, 1997 8 (pooled as 1 0.7 2,500
floating sample)
habitat
Cat Hai province Green Mussel, 1997 12 (pooled as 2 420
floating 1 sample)
habitat
Lach Truong, Green Mussel, 1997 33 (pooled as 1.2 610
Thanh Hoa aquaculture 1 sample)
Ron River estuary, Green Mussel, 1997 50 (pooled as 0.6 470
Ky Anh fishing 1 sample)
village
Lang Co, Hue Green Mussel 1997 143 (pooled as 0.9 34,000
1 sample)
Thi Nai, Binh Dinh Green Mussel, 1997 54 (pooled as 1.1 220
urban, shipping 1 sample)
traffi
fic,
aquaculture
Phan Ri estuary, Green Mussel, 1997 30 (pooled as 1.1 240
Phan Ri urban, fifishing 1 sample)
village
 Organic Pollutants in Vietnam 233

HCHs CHLs PCBs Reference Remarks

120 (47–210) 7.9 (<0.5–17) 580 (270–950) Kannan et DDTs: sum of p,p′-DDE,
al. 1995 p,p′-DDD and
48 (32–74) 8.9 (>0.5–12) 630 (160–1,300) Kannan et p,p′-DDT
al. 1995 HCHs: sum of α-, β-, and
110 (33–200) 3.2 (<0.5–18) 950 (190–3,100) Kannan et γ
γ-HCH
al. 1995 CHLs: sum of cis-
chlordane, trans-
chlordane,
cis-nonachlor, trans-
nonachlor and
oxychlordane
350 (27–210) 110 (29–210) 110 (81–140) Minh et al. PCBs: quantifified by an
2002 equivalent mixture of
Kanechlor perparations
(KC-300, KC-400, KC-
150 3.9 100 Minh et al. 500 and KC-600)
2002
3.6 14 86 Monirith et DDTs: sum of p,p′-DDE,
al. 2003 p,p′-DDD and
12 12 20 Monirith et p,p′-DDT
al. 2003 HCHs: sum of α-, β-, and
γ
γ-HCH
5.7 24 450 Monirith et CHLs: sum of cis-
al. 2003 chlordane, trans-
chlordane,
3 5 110 Monirith et cis-nonachlor, trans-
al. 2003 nonachlor and
oxychlordane
3.3 13 65 Monirith et PCBs: quantifified by an
al. 2003 equivalent mixture of
5.5 20 190 Monirith et Kanechlor perparations
al. 2003 (KC-300, KC-400, KC-
500 and KC-600)
10 — 380 Monirith et
al. 2003
6.3 36 26 Monirith et
al. 2003

2.9 11 80 Monirith et
al. 2003
234 T.B. Minh et al.

Table 3. (cont.)

Sample/sampling site Lipid

Location description Year n content (%) DDTs

Hanoi Clam (hyriopsis), 1996 15 (pooled as — 7,400

irrigation canal 1 sample)
Hanoi Carp (cyprinus 1996 3 (pooled as 1 — 22,000
carpio), rice field
fi sample)
Balat estuary, Thai Shrimp 1996 35 (pooled as — 1,000
Binh (metapenaeus) 1 sample)
Balat estuary, Thai Clam (Meretrix 1996 43 (pooled as — 830
Binh meretrix) 1 sample)
Diem Dien estuary, Clam (Mactra 1996 13 (pooled as — 1,000
Thaibinh quadrangularis), 1 sample)
intertidal mudfl flat
coastal area
Tra Co, Mong Cai, Clam (Meretrix 1997 1 pooled — 1,200
northern coast of meretrix), sample
Vietnam intertidal mudfl flat
areas
Mong Duong, Clam (Meretrix 1997 1 pooled — 590
northern coast of meretrix), sample
Vietnam intertidal mudfl flat
areas
Ha Long, Quang Clam (Meretrix 1997 1 pooled — 660
Ninh, northern meretrix), sample
coast of Vietnam intertidal mudfl flat
areas
Hai Phong, Clam (Meretrix 1997 1 pooled — 850
northern coast of meretrix). sample
Vietnam intertidal mudfl flat
areas
Ba Lat estuary, Clam (Meretrix 1997 1 pooled — 920
Thai Binh, meretrix), sample
northern coast of intertidal mudfl flat
Vietnam areas
Cau Dien, Nhue Freshwater snails 1997 1 pooled — 23,000
River, Hanoi (Angulyagra sp.), sample
densely populated
industrial
Cau Dien, Nhue Freshwater snails 1997 1 pooled — 45,000
River, Hanoi (Angulyagra sp.), sample
densely populated
industrial
transformer
production
Nhue River, suburb Freshwater snails 1997 1 pooled — 320
Hanoi (Angulyagra sp.), sample
rural area
 Organic Pollutants in Vietnam 235

HCHs CHLs PCBs Reference Remarks

97 — 480 Nhan et al. DDTs: sum of p,p′-DDE,

1998 p,p′-DDD and
370 — 2,600 Nhan et al. p,p′-DDT
1998 HCHs: sum of α-, β-, and
390 — 490 Nhan et al. γ
γ-HCH
1998 PCBs: sum 13 congeners
57 — 220 Nhan et al.
1998
30 — 200 Nhan et al.
1998

2,400 — 900 Nhan et al. DDTs: sum of p,p′-DDE

1999 and p,p′-DDD and
p,p′-DDT
HCHs: sum of α-, β-, and
400 — 480 Nhan et al. γ
γ-HCH
1999 PCBs: sum 13 congeners

93 — 470 Nhan et al.

1999

77 — 580 Nhan et al.

1999

71 — 250 Nhan et al.

1999

29 14 720 Nhan et al. DDTs sum of p,p′-DDE,

2001 p,p′-DDD, p,p′-DDT,
o,p′-DDE, p,p′-DDD,
p,p′-DDT and DDMU
21 27 2,200 Nhan et al. PCBs: sum 13 congeners
2001 HCHs: sum of α-, β-, and
γ
γ-HCH
CHLs: sum of cis-
chlordane, trans-
chlordane,
9.8 — 330 Nhan et al. trans-nanachlor and
2001 heptachlor
236 T.B. Minh et al.

Table 3. (cont.)

Sample/sampling site Lipid

Location description Year n content (%) DDTs

Hanoi downtown Freshwater snails 1997 1 pooled — 2,700

(Angulyagra sp.), sample
densely populated
industrial area
To Lich River, Freshwater snails 1997 1 pooled — 5,300
suburb Hanoi (Angulyagra sp.), sample
densely populated
industrial area
Red River, suburb Freshwater snails 1997 1 pooled — 910
Hanoi (Angulyagra sp.), sample
rural area
Gia Lam, suburb Freshwater snails 1997 1 pooled — 1,000
Hanoi (Angulyagra sp.), sample
rural area
Dong Anh, suburb Freshwater snails 1997 1 pooled — 640
Hanoi (Angulyagra sp.), sample
rural area
Mekong River, Can Catfi fish 2004 20 3.8 (0.6–7.2) 59 (7.9–
Tho (Pangasianodon 150)
hypophthalmus),
common
aquaculture
Can Tho Catfifish (Clarias sp.), 2004 5 3.6 (3.2–4.1) 390 (330–
from a pond near 700)
municipal
dumping site
Con Lu island, Red Resident birds 1997 16 1.9–16 6,200(1,100–
River estuary, (including 7 13,000)b
northern Vietnam species), wetland
wintering grounds
Con Lu island, Red Migratory birds 1997 84 4.1–33 2,900 (750–
River estuary, (including 17 6,800)
northern Vietnam species), wetland
wintering grounds

a
Mean (range).
b
Average concentrations of 7 resident and 17 migratory species.
 Organic Pollutants in Vietnam 237

HCHs CHLs PCBs Reference Remarks

13 1.8 1,000 Nhan et al.

2001

19 — 3,000 Nhan et al.

2001

8.2 2.2 480 Nhan et al.

2001

15 — 340 Nhan et al.

2001

11 1 280 Nhan et al.

2001

0.47 (<0.03–1.5) 0.62 (<0.01–2.6) 7.2 (0.91–27) Minh et al. DDTs: sum of p,p′-DDE,
2005, p,p′-DDD and
2006 p,p′-DDT
HCHs: sum of α-, β-, and
γ
γ-HCH
2.2 (0.86–5.1) 5.7 (4.2–8.2) 50 (37–77) Minh et al.
2005,
2006

150 (23–310) 100 (5–550) 780 (250–2,400) Minh et al. CHLs: sum of cis-
2002 chlordane, trans-
chlordane,
cis-nonachlor, trans-
330 (20–1,700) 22 (5.3–130) 530 (82–1,600) Minh et al. nonachlor and
2002 oxychlordane
PCBs: quantifified by an
equivalent mixture of
Kanechlor preparations
(KC-300, KC-400, KC-
500 and KC-600)
238 T.B. Minh et al.

Table 4. Concentrations of Persistent Organochlorines in Human Samples from

Vietnam.

Samples/sampling site
Locations Year despcription Unit n DDTs

Me Tri and Tu Liem, 2000 Breast milk, ng g−1 lipid 2,400

Hanoi primiparous women
near municipal
dumping sites

Me Tri and Tu Liem, 2000 Breast milk, ng g−1 lipid 1,700

Hanoi multiparous women
near municipal
dumping sites

Me Tri and Tu Liem, 2000 Breast milk, overall ng g−1 lipid 42 2,100 (480 –
Hanoi near municipal 6,900)a
dumping sites
Vinh Loc and Dong 2001 Breast milk, ng g−1 lipid 3,020
Thanh, Hochiminh primiparous women
City near municipal
dumping sites

Vinh Loc and Dong 2001 Breast milk, ng g−1 lipid 1,500
Thanh, Hochiminh multiparous women
City near municipal
dumping sites
Vinh Loc and Dong 2001 Breast milk, overall ng g−1 lipid 44 2,300 (440 –
Thanh, Hochiminh near municipal 17,000)
City dumping sites
Tan Than village, 1985–87 Breast milk, rural area ng g−1 lipid 2 10,500
suburb Hochiminh
City
Song Be province, 1985–87 Breast milk, rural area ng g−1 lipid 3 12,000
southern Vietnam

Hochiminh City 1985–87 Breast milk, urban ng g−1 lipid 7 11,400

area

Hanoi, rural area 1994 Serum ng mL−1 30 12 (1.2–59)

Hanoi, urban area 1994 Serum ng mL−1 8 32 (12–68)

Hanoi, breast cancer 1994 Serum ng mL−1 21 16 (1.2–52)

cases
Hanoi, control 1994 Serum ng mL−1 21 21 (1.5–88)
samples for breast
cancer cohort
survey

a
Mean (range).
 Organic Pollutants in Vietnam 239

HCHs CHLs HCB PCBs Reference Remark

69 2.5 4.2 76 Minh et al. DDTs: sum of p,p′-

2004 DDE, p,p′-DDD
and p,p′-DDT

HCHs: β-HCH only

46 1.4 3.5 72 Minh et al. CHLs: sum of
2004 oxychlordane,
trans-nonachlor
and
cis-nonachlor
58 (11–160) 2.0 (<0.72–13) 3.9 (0.62–9.5) 74 (26–210) Minh et al. PCBs: quantified
fi by
2004 an equivalent
mixture of
14 7.8 2.8 88 Minh et al. Kanechlor
2004 preparations
(KC-300, KC-
400, KC-500 and
KC-600)
13 6 2.1 70 Minh et al.
2004

14 (4.1–35) 6.9 (1.3–2.6) 2.5 (1.3–10) 79 (29–200) Minh et al.

2004

25 <2.0 <2.0 49 Schecter et DDTs: sum of p,p′-

al. 1989a DDE and
p,p′-DDT
37 <2.0 10 28 Schecter et HCHs: sum of α-,
al. 1989a β- and γ-HCH;
γ
CHLs:
oxychlordane on
250 3 3 84 Schecter et PCBs: sum of CB-
al. 1989a 138, CB-153 and
CB-180
— — — — Schecter et DDTs: sum of p,p′-
al. 1997 DDE and
p,p′-DDT
— — — — Schecter et
al. 1997
— — — — Schecter et
al. 1997
— — — — Schecter et
al. 1997
240 T.B. Minh et al.

preliminary survey to examine the levels of PBDEs in common aquaculture

catfish
fi from Can Tho, a city located on the Mekong River, in southern
Vietnam (Minh NH et al. 2006b). Catfish fi from a pond located near a
municipal waste site of Can Tho were also analyzed to evaluate the role of
dumping site as a source of pollution. Interestingly, concentrations of
PBDEs and persistent OCs such as PCBs, DDTs, CHLs, and HCHs in the
dumping site fish
fi were signifi ficantly higher than those in fish collected from
a common aquaculture site (Fig. 9, Minh NH et al. 2006b), suggesting an
additional exposure of the dumpsite catfish
fi to PBDEs. In the dumping site,
municipal wastes including household goods and small electric appliances,
that may contain PBDEs as fl flame retardants, were dumped. Under ambient
conditions, PBDEs may be emitted from such materials and consequently
contaminate the dumping site soil. Therefore, it is anticipated that runoff
and leaching water from the dumping site during flood and rains may have
carried PBDEs to other vicinities, causing higher contamination in the
catfish.
fi Higher levels in fish from this study further highlighted the role of
open dumping sites for municipal wastes as a conspicuous source of various
toxic chemicals, including a new group of POPs such as PBDEs.
To our knowledge, there are very few studies on OC contamination
in higher trophic animals from Vietnam. A survey conducted in 1997
determined OC concentrations in resident and migratory birds from North
Vietnam (Minh TB et al. 2002). Resident birds contained higher
concentrations of DDTs than those in migrants (Fig. 10); this indicates
recent exposure to DDTs in resident birds from North Vietnam, where
elevated DDT contamination is very common, as discussed earlier.
Interestingly, accumulation of HCHs revealed a contrasting pattern,
showing apparently greater concentration in migratory birds (Fig. 10),

DDTs

Common catfish
HCHs
Dumping site catfish

PCBs

Mean Max
PBDEs

0.1 1 10 100 1000

Concentration (ng/g lipid)

Fig. 9. Comparison of PBDE and persistent organochlorine residues in catfish

fi from
open dumping sites and in common aquaculture catfi
fish from Can Tho City, Mekong
River, southern Vietnam (Minh NH et al. 2006b).
Fig. 10. Accumulation of persistent organochlorines in birds from Vietnam accord-
ing to migratory behavior (Minh TB et al. 2002).

which could be the result of accumulation in stopover sites during migration

in some polluted areas such as India, southern China, and Japan. The role
of these countries as potential sources of HCH accumulation in wintering
migratory birds breeding in Lake Baikal, Russia, has also been suggested
in a recent study (Kunisue et al. 2002). Concentrations of PCBs were similar
in residents and migratory species and levels were relatively low, indicating
a smaller source of PCBs in North Vietnam in recent years. Thus,
accumulation pattern of OCs in birds from North Vietnam according to
their migratory behavior reflects
fl the status of each contaminant. This
phenomenon also suggests the suitability of using birds as bioindicators for
monitoring POPs in the environment.
International comparison of OC residues in fish,
fi mussels, and birds from
Vietnam indicated relatively higher levels of DDTs in Vietnamese samples
(Fig. 11). Results of recent surveys in the Asia-Pacific fi Mussel Watch
Program showed that DDT levels in Vietnamese mussels were lower than
in those in southern China and Hong Kong, but higher than those in
samples collected from most of the other countries in the East Asian region
(Fig. 11; Monirith et al. 2003). Kannan et al. (1995) reported mean
concentrations of DDT in fish from Vietnam were the highest when com-
pared with India, Thailand, Indonesia, and Australia. Contamination
patterns were similar for birds, which showed that DDT levels in some
resident species in Vietnam were among the highest values reported for the
Asian countries surveyed (see Fig. 11; Minh TB et al. 2002). Although the
recent amounts of DDT used in Vietnam were lower than those of other
countries in the region, the extent of DDT contamination in environmental
samples in Vietnam is higher. This observation suggests that the application
242 T.B. Minh et al.

Fig. 11. Comparison of DDTs residues in fi fish, mussels, and birds in Asia-Pacifi fic
countries. [From: fifish: Kannan et al. (1995), Nakata et al. (1995), Guruge et al.
(1997), Monirith et al. (2000), Minh TB et al. (2002); mussels: Monirith et al. (2003);
birds: Hoshi et al. (1998), Choi et al. 2001, Kunisue et al. (2002, 2003), Minh TB
et al. (2002), Tanabe et al. (1998)].

of DDT in Vietnam has continued until very recently, resulting in elevated

contamination of these compounds in different species occupying low to
high trophic levels in the food chain.

C. Foodstuffs
The first examination of OC contamination in foodstuffs from Vietnam was
conducted by Schecter et al. (1990a). Pork and chicken fat were analyzed,
and relatively high levels of DDTs were found (190–5100 ng/g lipid).
However, residues levels of other contaminants such as HCHs, CHLs,
HCB, and PCBs were relatively low.
A subsequent investigation done by Kannan et al. (1992) in the samples
collected during January 1990 and May 1991 examined various kinds of
foodstuffs including rice, pulses, oil, butter, animal fat, meat, fish,
fi and
seafood from Hanoi, northern Vietnam, and Tay Ninh and Hochiminh City,
southern Vietnam. The results demonstrated significant
fi concentrations of
PCBs in foods from both the northern and southern parts, which is consistent
 Organic Pollutants in Vietnam 243

with the data reported for soils collected at the same locations (Thao et al.
1993a,b). DDT concentrations were prominent in all kinds of foodstuffs,
particularly in oil and animal fat. The ongoing application of DDT at the
time of this investigation was also evident. In particular, composition of
DDT compounds showed that p,p′-DDT was the predominant isomer in
rice, pulses, and oils, accounting for approximately 50% of the total DDT
concentrations (Kannan et al. 1992). Very recently, Schecter et al. (2003)
reported some alarmingly elevated concentrations of PCDD/Fs in foodstuffs
from Bien Hoa City, southern Vietnam, where Agent Orange was sprayed
during the Vietnam War. Several samples of duck, toad, chicken, and
snakehead fish collected from the former U.S. Army base site contained
elevated concentrations of DDTs. The high levels of DDTs found in meat
of livestock and fish from these studies indicate the application of technical
DDT to prevent malaria near cattle sheds, stagnant water bodies, and
drainage channels, which are common breeding sites for mosquitoes
(Kannan et al. 1992).

D. Human Exposure
Available data on human exposure to organochlorine insecticides and
PCBs in Vietnamese residents are limited. A preliminary survey by Schecter
et al. (1989a) reported very high concentrations of DDTs and HCHs in
human breast milk from some locations in and around Hochiminh. DDT
levels were higher in both rural and urban areas, ranging from 10,500 to
12,000 ng/g lipid. PCB levels, however, were relatively low compared to
other countries. A subsequent study conducted in 2000–2001 provided
more extensive data on human exposure and insights into the accumulation
kinetics of PCBs and OC insecticides such as DDTs, HCHs, CHLs, HCB,
and tris(4-chlorophenyl)methane (TCPMe), a newly detected environmental
contaminant that exhibits weak endocrine-disrupting properties. Breast
milk from 96 nursing women living near the dumping sites of municipal
wastes in Hanoi and Hochiminh, the two largest metropolitan cities in
Vietnam, was analyzed for these contaminants (see Table 4). In general,
similar degrees of exposure to DDTs, CHLs, HCB, and PCBs were observed
among samples from Hanoi and Hochiminh. Interestingly, HCH residues
in breast milk of women from Hanoi were signifi ficantly higher than those
in Hochiminh, suggesting recent high background levels of HCHs in the
northern compared to the southern region. Consistent results where also
observed in the survey on sediment and bivalves from various locations
along the northern coast of Vietnam, which demonstrated relatively higher
residues in sites near the China border (Nhan et al. 1998, 1999). Earlier
reports also pointed out similar spatial distribution in different kinds of
environmental samples, showing higher levels of HCHs in Hanoi compared
to Hochiminh (Thao et al. 1993a; Iwata et al. 1994; Kannan et al. 1995). In
addition to the influence
fl of the possible transport from China, the world
244 T.B. Minh et al.

leading HCH user, differences in climate between Hanoi and Hochiminh

could be an alternative explanation. The Mekong River delta in southern
Vietnam is characterized by the typical tropical climate with high
temperatures and heavy rainfall. Rapid volatilization of highly volatile
HCH isomers may therefore be enhanced in southern Vietnam, resulting
in lower residues in various environmental and human samples.
Similar to those observed in environmental samples, human exposures
to DDTs in Vietnam were among the highest in the developing as well
as developed nations. As discussed earlier, high DDT contamination in
Vietnam has been apparent in many environmental samples. This is also
the case for humans, which raises concern over the possible impacts on
human health. These results suggest that Vietnam is a potential source of
DDTs in the South Asian region.

E. Dioxin Contamination
Southern Vietnam has long been considered as a well-known region where
Agent Orange was extensively sprayed during 1961–1971 in the Vietnam
War, resulting in severe dioxin contamination of various environmental
media and the food chain, including humans. During the past three decades,
Schecter and coworkers have been conducting investigations on dioxin
contamination in southern Vietnam, including sediments, foodstuffs, and,
particularly, samples from humans living near the “hot spot” of dioxin
contamination (Table 5). In general, dioxin contamination as a result of
Agent Orange can be characterized by the predominant of 2,3,7,8-

Table 5. Concentration of PCDD/Fs and Dioxin-Like PCBs in Different

Environmental Samples from Vietnam.

Sample/sampling
Location Year site description Unit n PCDDs

Soil, sediment
Saigon River 1984/85 Sediment, pg g−1 dry wt 1 5,800
industrial area
Dong Nai River 1984/85 Sediment, near pg g−1 dry wt 5 1,200 (830–
Orange Agent 1,400)
spraying during
Vietnam War
Red River 1984/85 Sediment, river pg g−1 dry wt 2 240 (220–
250)
Bien Hoa 2000 Soil, near the Air ng g−1 dry wy 4 590 (0.039–
Base, former 1,645)
Agen Orange
Storage
 Organic Pollutants in Vietnam 245

tetrachlorodibenzo-p - -dioxin (2,3,7,8-TCDD), the major contaminants of

the herbicide 2,4,5-T, a constituent of Agent Orange. 2,3,7,8-TCDD is one
of the most toxic congeners and has received considerable attention.
Research has shown that this congener can caused large number of adverse
health effects in animals (ATSDR 1998). As seen in Table 5, 2,3,7,8-TCDD
was the major component, and its concentrations were higher in foods and
human samples including blood, adipose tissues, and breast milk.
Dwernychuk et al. (2002) surveyed soils, sediments, and foods from
Aluoi valley, a “hot spot” of Agent Orange spraying and former U.S. Army
base sites. The highest concentrations of PCDD/Fs in soil were 2200 pg/g
dry wt, which is much higher than those in background levels in many
industrialized countries. Of greater concern, however, are high levels of
PCDD/Fs in breast milk, a major dietary intake for breast-fed children. A
survey throughout the valley indicates that breast-fed infants of primipara
groups had intake values 27 fold exceeding the Tolerable Daily Intake
(TDI) proposed by WHO. This fact highlights a new environmental issue:
that the fi
first child is exposed to high levels of contaminants and will always
be at higher risk. Breast-feeding has been highly recommended by various
organizations and scientists because of the benefits fi for children, such as
passing antibodies to the infants and reducing the risk of allergic reactions.
However, for communities with high exposure to toxic contaminants such
as people in underdeveloped countries such as Vietnam, breast-feeding
unfortunately has become an effective route of transfer of toxic contaminants
to the next generation. Scientificfi and social efforts are therefore urgently
needed to mitigate exposure to reduce body burdens of dioxins in both
adults and children.

PCDFs Total PCDD/Fs Dioxin-like PCBs Reference Remark

1,000 6,800 — Schecter et al.

1989b
54 (5–140) 1,300 (840–1,500) — Schecter et al.
1989b

4.2 (3.4–4.9) 240 (220–255) — Schecter et al.

1989b
93 (39–147) 680 (39–1,790) — Schecter et al.
2001
246 T.B. Minh et al.

Table 5. (cont.)

Sample/sampling
Location Year site description Unit n PCDDs

Bien Hung Lake, 2000 Sediment, former pg g−1 dry wt 3 340 (199–
Bien Hoa Air Base 532)
Bien Hung Lake, 2000 Sediment, close pg g−1 dry wt 3 1,640 (1,410–
Bien Hoa to the former 1,970)
Air Base,
Agent Orange
Storage
Bien Hung Lake, 2000 Sediment, pg g−1 dry wt 2 600 (500–
Bien Hoa downstream 700)
and upstream
Dong Nai River, 2000 Sediment, pg g−1 dry wt 2 630 (540–
southern reservoir of 715)
Vietnam Bien Hung
Lake
Hanoi 2000 Sediment, control pg g−1 dry wt 1 400
samples for
samples near
former Air
Base
Aso, Aluoi valey, 1996 Soil, former US pg g−1 dry wt 1 650
Hue Army Force
Base
Aso, Aluoi valey, 1996 Sediment, pond, pg g−1 dry wt 1 500
Hue former US
Army Force
base
Aso, Aluoi valey, 1997 Soil, former US pg g−1 dry wt 1 1,600
Hue Army Force
Base
Aso, Aluoi valey, 1997 Soil, former US pg g−1 dry wt 1 880
Hue airstrip
Aso, Aluoi valey, 1997 Soil, manioc pg g−1 dry wt 1 170
Hue fi
fields
Aso, Aluoi valey, 1997 Sediment, pond, pg g−1 dry wt 4 150 (32–250)
Hue former US
Army Force
base
Aso, Aluoi valey, 1999 Soil, former US pg g−1 dry wt 10 360 (98–730)
Hue Army Force
Base
Tabat, Aluoi 1999 Soil, former US pg g−1 dry wt 9 740 (430–
valey, Hue Army Force 1,100)
Base
Aluoi, Aluoi 1999 Soil, former US pg g−1 dry wt 9 270 (110–
valey, Hue Army Force 430)
Base
Adot, Aluoi 1999 Soil, areas pg g−1 dry wt 3 350 (48–550)
valey, Hue sprayed with
Agent Orange
 Organic Pollutants in Vietnam 247

PCDFs Total PCDD/Fs Dioxin-like PCBs Reference Remark

10 (9–11) 1,050 (208–542) — Schecter et al.

2001
107 (89–134) 1,720 (1,510–2,100) — Schecter et al.
2001

24 (19–29) 625 (720–530) — Schecter et al.

2001

4.4 (2.0–6.8) 630 (544–720) — Schecter et al.

2001

70 470 — Schecter et al.

2001

92 740 — Dwernychuk et
al. 2002

5.6 510 — Dwernychuk et

al. 2002

98 1,700 — Dwernychuk et
al. 2002

80 960 — Dwernychuk et
al. 2002
7.1 180 — Dwernychuk et
al. 2002
2.7 (1.1–3.7) 150 (33–250) — Dwernychuk et
al. 2002

33 (5.6–75) 390 (100–810) — Dwernychuk et

al. 2002

14 (5.2–29) 750 (440–1,100) — Dwernychuk et

al. 2002

8.9 (5.0–19) 280 (120–440) — Dwernychuk et

al. 2002

2.7 (2–3.1) 350 (50–550) — Dwernychuk et

al. 2002
248 T.B. Minh et al.

Table 5. (cont.)

Sample/sampling
Location Year site description Unit n PCDDs

Huonglam, Aluoi 1999 Soil, areas pg g−1 dry wt 2 150 (72–220)

valey, Hue sprayed with
Agent Orange
Huongphong, 1999 Soil, areas pg g−1 dry wt 2 260 (240–
Aluoi valey, sprayed with 280)
Hue Agent Orange
Phuvinh, Aluoi 1999 Soil, areas pg g−1 dry wt 2 600 (560–
valey, Hue sprayed with 630)
Agent Orange
Hongthuong, 1999 Soil, areas pg g−1 dry wt 1 2,200
Aluoi valey, sprayed with
Hue Agent Orange
Bodot market, 1999 Soil, areas pg g−1 dry wt 1 1,300
Aluoi valey, sprayed with
Hue Agent Orange
Sonthuy, Aluoi 1999 Soil, areas pg g−1 dry wt 2 1,700 (1,200–
valey, Hue sprayed with 2,100)
Agent Orange
Hongquang, 1999 Soil, areas pg g−1 dry wt 1 88
Aluoi valey, sprayed with
Hue Agent Orange
Aluoi, Aluoi 1999 Soil, areas pg g−1 dry wt 1 46
valey, Hue sprayed with
Agent Orange
Aluoi market, 1999 Soil, areas pg g−1 dry wt 1 830
Aluoi valey, sprayed with
Hue Agent Orange
Hongkim, Aluoi 1999 Soil, areas pg g−1 dry wt 1 87
valey, Hue sprayed with
Agent Orange
Hongvan, Aluoi 1999 Soil, areas pg g−1 dry wt 2 110 (70–140)
valey, Hue sprayed with
Agent Orange
Taymo, Hanoi 2000 Soil, open pg g−1 dry wt 9 3,100
dumping site
for
municipal wastes TEQ (pg g−1 31
dry wt)
Taymo, Hanoi 2000 Soil, paddy fi
field, pg g−1 dry wt 1 360
control sites
for
Taymo open TEQ (pg g−1 0.94
dumping site dry wt)
Dongthanh, 2001 Soil, open pg g−1 dry wt 8 320
Hochiminh dumping site
City for
municipal wastes TEQ (pg g−1 1.4
dry wt)
 Organic Pollutants in Vietnam 249

PCDFs Total PCDD/Fs Dioxin-like PCBs Reference Remark

8.7 (1.3–16) 160 (73–240) — Dwernychuk et

al. 2002

6.1 (3.9–8.2) 270 (240–290) — Dwernychuk et

al. 2002

6.4 (4.3–8.5) 610 (560–640) — Dwernychuk et

al. 2002

5.5 2,210 — Dwernychuk et

al. 2002

110 1,400 — Dwernychuk et

al. 2002

59 (7.1–110) 1,800 (1,200–2,200) — Dwernychuk et

al. 2002

6 94 — Dwernychuk et
al. 2002

3.3 49 — Dwernychuk et
al. 2002

14 840 — Dwernychuk et
al. 2002

2.1 89 — Dwernychuk et
al. 2002

2.6 (2.3–2.8) 110 (72–140) — Dwernychuk et

al. 2002

2,900 6,000 (125–50,500) 2,600 (59–7,100) Minh et al. 2003

64 95 (0.4–850) 7.3 (0.22–59) Minh et al. 2003

11 370 130 Minh et al. 2003

0.08 1 0.097 Minh et al. 2003

47 370 (21–880) 910 (65–2,000) Minh et al. 2003

0.8 2.2 (0.02–4.4) 0.49 (0.01–1.0) Minh et al. 2003

250 T.B. Minh et al.

Table 5. (cont.)

Sample/sampling
Location Year site description Unit n PCDDs

Dongthanh, 2001 field, pg g−1 dry wt

Soil, paddy fi 3 160
Hochiminh control sites
City for
DongThanh open TEQ (pg g−1 0.8
dumping site dry wt)
Foodstuff/Biota
Hanoi 1985–87 Pork TEQ (pg g−1 2 —
wet wt)
Hanoi 1985–87 Pork fat TEQ (pg g−1 1 —
wet wt)
Hanoi 1985–87 Catfish
fi TEQ (pg g−1 1 —
wet wt)
Hanoi 1985–87 Carp TEQ (pg g−1 1 —
wet wt)
Hanoi 1985–87 Eel TEQ (pg g−1 1 —
wet wt)
Hanoi 1985–87 Cow fat TEQ (pg g−1 1 —
wet wt)
Hanoi 1985–87 Chicken fat TEQ (pg g−1 1 —
wet wt)
Southern 1985–87 Pork fat TEQ (pg g−1 1 —
Vietnam wet wt)
Southern 1985–87 Pork fat TEQ (pg g−1 1 —
Vietnam wet wt)
Song Be, 1985–87 Pork fat TEQ (pg g−1 1 —
southern wet wt)
Vietnam
Song Be, 1985–87 Chicken fat TEQ (pg g−1 1 —
southern wet wt)
Vietnam
Tan Uyen, 1985–87 Chicken egg TEQ (pg g−1 1 —
southern wet wt)
Vietnam
Tan Uyen, 1985–87 Fish TEQ (pg g−1 1 —
southern wet wt)
Vietnam
Tan Than, 1985–87 Fish TEQ (pg g−1 1 —
southern wet wt)
Vietnam
Southern 1985–87 Channa fish
fi TEQ (pg g−1 1 —
Vietnam muscle wet wt)
Southern 1985–87 Channa fish
fi TEQ (pg g−1 1 —
Vietnam muscle wet wt)
Southern 1985–87 Channa fish
fi liver TEQ (pg g−1 1 —
Vietnam wet wt)
Southern 1985–87 Clarius fi
fish TEQ (pg g−1 1 —
Vietnam muscle wet wt)
Binh Tri Thien, 1985–87 Catfish
fi TEQ (pg g−1 1 —
middle wet wt)
Vietnam
 Organic Pollutants in Vietnam 251

PCDFs Total PCDD/Fs Dioxin-like PCBs Reference Remark

23 180 (130–260) 160 (46–300) Minh et al. 2003

0.35 1.15 (0.36–1.2) 0.12 (0.027– Minh et al. 2003

0.19)

— 2.1 (1.9–2.2) — Schecter et al.

1989c
— 0.58 — Schecter et al.
1989c
— 0.26 — Schecter et al.
1989c
— 0.64 — Schecter et al.
1989c
— 0.35 — Schecter et al.
1989c
— 3.5 — Schecter et al.
1989c, 1989d
— 3.3 — Schecter et al.
1989c, 1989d
— 10.9 — Schecter et al.
1989c
— 4.1 — Schecter et al.
1989c
— 3.5 — Schecter et al.
1989c, 1989d

— 32 — Schecter et al.
1989c, 1989d

— 1.6 — Schecter et al.

1989c

— 0.53 — Schecter et al.

1989c

— 0.82 — Schecter et al.

1989c

— 0.17 — Schecter et al.

1989c
— 0.26 — Schecter et al.
1989c
— 4.2 — Schecter et al.
1989c
— 0.36 — Schecter et al.
1989c
— 5.7 — Schecter et al.
1989c
252 T.B. Minh et al.

Table 5. (cont.)

Sample/sampling
Location Year site description Unit n PCDDs

Hochiminh City 1985–87 Eel TEQ (pg g−1 2 —

wet wt)
Hochiminh City 1985–87 Fish TEQ (pg g−1 1 —
wet wt)
Hochiminh City 1985–87 Fish TEQ (pg g−1 1 —
wet wt)
Hochiminh City 1985–87 Fish TEQ (pg g−1 1 —
wet wt)
Hochiminh City 1985–87 Crab TEQ (pg g−1 1 —
wet wt)
Hochiminh City 1985–87 Crab meat TEQ (pg g−1 1 —
wet wt)
Hochiminh City 1985–87 Chicken egg TEQ (pg g−1 1 —
wet wt)
Hochiminh City 1985–87 Duck egg TEQ (pg g−1 1 —
wet wt)
Southern 1985–87 Turtle fat TEQ (pg g−1 1 —
Vietnam wet wt)
Southern 1985–87 Turtle muscle TEQ (pg g−1 1 —
Vietnam wet wt)
Southern 1985–87 Turtle liver TEQ (pg g−1 1 —
Vietnam wet wt)
Southern 1985–87 Turtle gall TEQ (pg g−1 1 —
Vietnam baldder wet wt)
Southern 1985–87 Turtle ovaries TEQ (pg g−1 1 —
Vietnam wet wt)
Southern 1985–87 Snake TEQ (pg g−1 1 —
Vietnam wet wt)
Hochiminh City 1988/89 Pork stick pg g−1 lipid 1 39

Hochiminh City 1988/89 Pork fat pg g−1 lipid 1 41

Hochiminh City 1988/89 Chicken liver pg g−1 lipid 1 94

Hochiminh City 1988/89 Chicken fat pg g−1 lipid 1 44

Bienhoa City, 2002 Snake head pg g−1 lipid 1 16,000

southern (channa
Vietnam striata), nearby
Orange Agent
Air Base
Bienhoa City 2002 Climbing perch pg g−1 lipid 1 13
(anabas
testudineus),
nearby Orange
Agent Air
Base
 Organic Pollutants in Vietnam 253

PCDFs Total PCDD/Fs Dioxin-like PCBs Reference Remark

— 0.05 (0.04–0.07) — Schecter et al.

1989c
— 0.05 — Schecter et al.
1989c
— 0.9 — Schecter et al.
1989c
— 0.28 — Schecter et al.
1989c
— 2.6 — Schecter et al.
1989c
— 0.52 — Schecter et al.
1989c
— 0.55 — Schecter et al.
1989c
— ND — Schecter et al.
1989c
— 1.8 — Schecter et al.
1989c
— 2.6 — Schecter et al.
1989c, 1989d
— 22 — Schecter et al.
1989c, 1989d
— 6.5 — Schecter et al.
1989c, 1989d
— 86 — Schecter et al.
1989c,1989d
— 13 — Schecter et al.
1989c
4.3 43 — Schecter et al.
1990a
3.5 45 — Schecter et al.
1990a
58 150 — Schecter et al.
1990a
5.1 49 — Schecter et al.
1990a
200 16,200 119,000 Schecter et al.
2003

15 28 4,500 Schecter et al.

2003
254 T.B. Minh et al.

Table 5. (cont.)

Sample/sampling
Location Year site description Unit n PCDDs

Bienhoa City 2002 Catfi

fish (clarias pg g−1 lipid 1 4.4
fuscus), nearby
Orange Agent
Air Base
Bienhoa City 2002 Catfish
fi (clarias pg g−1 lipid 1 6.3
fuscus), nearby
Orange Agent
Air Base
Bienhoa City 2002 Carp (ostechilus pg g−1 lipid 1 41
hasselti),
nearby Orange
Agent Air
Base
Bienhoa City 2002 Duck, nearby pg g−1 lipid 1 560
Orange Agent
Air Base
Bienhoa City 2002 Duck, nearby pg g−1 lipid 1 550
Orange Agent
Air Base
Bienhoa City 2002 Toad, nearby pg g−1 lipid 1 22,000
Orange Agent
Air Base
Bienhoa City 2002 pork, nearby pg g−1 lipid 1 2.5
Orange Agent
Air Base
Bienhoa City 2002 pork nearby pg g−1 lipid 1 3.6
Orange Agent
Air Base
Bienhoa City 2002 Beef, nearby pg g−1 lipid 1 6.7
Orange Agent
Air Base
Bienhoa City 2002 Beef, nearby pg g−1 lipid 1 7.2
Orange Agent
Air Base
Bienhoa City 2002 Chicken, nearby pg g−1 lipid 1 400
Orange Agent
Air Base
Bienhoa City 2002 Chicken, nearby pg g−1 lipid 1 5.9
Orange Agent
Air Base
Bienhoa City 2002 Chicken, nearby pg g−1 lipid 1 11
Orange Agent
Air Base
Bienhoa City 2002 Chicken, nearby pg g−1 lipid 1 450
Orange Agent
Air Base
Human samples
Nine hospital in 1984 Adipose tissues pg g−1 lipid 9 150 (9.2–240)
Hanoi
 Organic Pollutants in Vietnam 255

PCDFs Total PCDD/Fs Dioxin-like PCBs Reference Remark

5.1 9.5 1,200 Schecter et al.

2003

17 23 3,300 Schecter et al.

2003

13 54 7,200 Schecter et al.

2003

40 600 3,300 Schecter et al.

2003

41 590 2,800 Schecter et al.

2003

2,600 25,000 960,000 Schecter et al.

2003

0.21 2.7 1,300 Schecter et al.

2003

11 15 450 Schecter et al.

2003

1.5 8.2 580 Schecter et al.

2003

1.3 8.5 4.6 Schecter et al.

2003

45 450 40,000 Schecter et al.

2003

26 32 34 Schecter et al.
2003

1.4 12 51 Schecter et al.

2003

42 490 39,000 Schecter et al.

2003

35 (5.3–47) 190 (15–290) — Schecter et al. 2,3,7,8-substituted

1986 congeners only
256 T.B. Minh et al.

Table 5. (cont.)

Sample/sampling
Location Year site description Unit n PCDDs

Hochiminh City 1984 Adipose tissues pg g−1 lipid 15 1,600 (190–

hospital 4,700)
Seven areas in 1973 Breast milk, pg g−1 lipid 9 680 (220–
south Vietnam Orange Agent 1,500)
sprayed sites,
Vietnam War
has not ended
Hochiminh City 1985 Breast milk, pg g−1 lipid 4 12 (10–13)
Children
Hospital
Hochiminh City 1985 Breast milk, pg g−1 lipid 3 <3–12
Obstetrical and
Gynecological
Hospital
Hanoi 1985 Breast milk, pg g−1 lipid 2 <2.0
Swedish
Hospital
Dongnai River, 1985 Breast milk, near pg g−1 lipid 2 <23
southern Tan Uyen
Vietnam village,
Dongnai River
Tan Than village, 1985–87 Breast milk, rural pg g−1 lipid 2 510
southern area
Vietnam
Song Be 1985–87 Breast milk, rural pg g−1 lipid 3 580
Hospital, area
southern
Vietnam
Pediatric 1985–87 Breast milk, pg g−1 lipid 7 760
Hospital, urban area
Hochiminh
City
Obstetrical and 1985–87 Adipose tissue pg g−1 lipid 3 6.4 (4.4–9.6)
Gynecological
Hospital,
Hochiminh City
Binh Dan 1985–87 Adipose tissue pg g−1 lipid 5 8.8 (1.2–19)
Hospital,
Hochiminh
City
Dong Nai 1985–87 Adipose tissue pg g−1 lipid 2 10 (7.6–13)
Provincial
Hospital,
southern
Vietnam
Hue provincial 1985–87 Adipose tissue pg g−1 lipid 3 16 (9.6–26)
Hospital,
middle
Vietnam
 Organic Pollutants in Vietnam 257

PCDFs Total PCDD/Fs Dioxin-like PCBs Reference Remark

110 (30–280) 1,700 (220–5,000) — Schecter et al. 2,3,7,8-substituted

1986 congeners only
62 (<2–190) 740 (220–1,700) — Schecter et al.
1987

— 12 (10–13) — Schecter et al. 2,3,7,8-TCDD

1987 only

— <3–12 — Schecter et al. 2,3,7,8-TCDD

1987 only

— <2.0 — Schecter et al. 2,3,7,8-TCDD

1987 only

— <23 — Schecter et al. 2,3,7,8-TCDD

1987 only

140 650 — Schecter et al.

1989a

100 680 — Schecter et al.

1989a

260 1,020 — Schecter et al.

1989a

— 6.4 (4.4–9.6) — Schecter et al. 2,3,7,8-TCDD

1989e only

— 8.8 (1.2–19) — Schecter et al. 2,3,7,8-TCDD

1989e only

— 10 (7.6–13) — Schecter et al. 2,3,7,8-TCDD

1989e only

— 16 (9.6–26) — Schecter et al. 2,3,7,8-TCDD

1989e only
258 T.B. Minh et al.

Table 5. (cont.)

Sample/sampling
Location Year site description Unit n PCDDs

Hochiminh City 1984–85 Adipose tissue pg g−1 lipid 2 1,045 (390–

1,700)
Soc Trang, 1984–85 Adipose tissue pg g−1 lipid 1 1,600
southern
Vietnam
Vung Tau, 1984–85 Adipose tissue pg g−1 lipid 1 4,700
southern
Vietnam
Tayninh,southern 1984–85 Adipose tissue, pg g−1 lipid 1 4,400
Vietnam resident close
to Agent
Orange spray
areas
Nha Be, southern 1984–85 Adipose tissue pg g−1 lipid 1 13
Vietnam
Long An, 1984–85 Adipose tissue pg g−1 lipid 1 7
southern
Vietnam
O Mon, Hau 1984–85 Adipose tissue pg g−1 lipid 1 4
Giang,
southern
Vietnam
Cuu Long, 1984–88 Adipose tissue pg g−1 lipid 1 410
southern
Vietnam
Dong Nai, 1984–88 Adipose tissue pg g−1 lipid 1 190
southern
Vietnam
Lam Dong, 1984–88 Adipose tissue pg g−1 lipid 1 1,150
southern
Vietnam
Binh Tri Thien, 1984–88 Adipose tissue pg g−1 lipid 1 300
middle
Vietnam
Tay Ninh, 1984–88 Adipose tissue pg g−1 lipid 1 1,310
southern
Vietnam
Tien Giang, 1984–88 Adipose tissue pg g−1 lipid 1 930
southern
Vietnam
Song Be, 1984–88 Adipose tissue pg g−1 lipid 1 2,010
southern
Vietnam
Phu Khanh, 1984–88 Adipose tissue pg g−1 lipid 1 3,000
southern
Vietnam
Hanoi 1980s Breast milk pg g−1 lipid 1 (28 93
samples
pooled)
 Organic Pollutants in Vietnam 259

PCDFs Total PCDD/Fs Dioxin-like PCBs Reference Remark

49 (39–58) 1,100 (430–1,760) — Phuong et al.

1989
130 1,730 — Phuong et al.
1989

31 4,730 — Phuong et al.

1989

160 4,560 — Phuong et al.

1989

— 13 — Phuong et al. 2,3,7,8-TCDD

1989 only
— 7 — Phuong et al. 2,3,7,8-TCDD
1989 only

— 4 — Phuong et al. 2,3,7,8-TCDD

1989 only

71 480 — Phiet et al. 1989

36 230 — Phiet et al. 1989

195 1,350 — Phiet et al. 1989

100 400 — Phiet et al. 1989

88 1,400 — Phiet et al. 1989

76 1,000 — Phiet et al. 1989

150 2,160 — Phiet et al. 1989

280 3,300 — Phiet et al. 1989

23 120 — Schecter et al.

1989f
260 T.B. Minh et al.

Table 5. (cont.)

Sample/sampling
Location Year site description Unit n PCDDs

Hochiminh City 1980s Breast milk pg g−1 lipid 1 (38 300

samples
pooled)
Song Be, 1980s Breast milk pg g−1 lipid 1 (12 280
southern samples
Vietnam pooled)
Tan Uyen, 1980s Breast milk pg g−1 lipid 3 pooled 640 (160–
southern 310)
Vietnam
Can Gio, 1980s Breast milk pg g−1 lipid 1 (3 94
southern samples
Vietnam pooled)
Long Xuyen, 1980s Breast milk pg g−1 lipid 1 (2 48
southern samples
Vietnam pooled)
Hochiminh City 1980s Breast milk pg g−1 lipid 1 (15 390
samples
pooled)
Hochiminh City 1980s Breast milk pg g−1 lipid 1 (8 240
samples
pooled)
Song Be, 1980s Breast milk pg g−1 lipid 1 (12 280
southern samples
Vietnam pooled)
Various 1980s Adipose tissues pg g−1 lipid 25 19 (7.7–36)
provinces in
Mekong River
Delta,
southern
Vietnam
Binh Long, 1988–89 Breast milk pg g−1 lipid 1 (4 215
southern samples
Vietnam pooled)
Vung Tau, 1988–89 Breast milk pg g−1 lipid 1 (5 280
southern samples
Vietnam pooled)
Tay Ninh, 1988–89 Breast milk pg g−1 lipid 1 (4 580
southern samples
Vietnam pooled)
Song Be, 1988–89 Breast milk pg g−1 lipid 1 (4 124
southern samples
Vietnam pooled)
Hanoi 1988–89 Adipose tissue pg g−1 lipid 1 (10 274
samples
pooled)
Various 1988–89 Adipose tissue pg g−1 lipid 13 596
provinces,
southern
Vietnam
 Organic Pollutants in Vietnam 261

PCDFs Total PCDD/Fs Dioxin-like PCBs Reference Remark

34 330 — Schecter et al.

1989f

47 330 — Schecter et al.

1989f

72 (11–190) 710 (170–500) — Schecter et al.

1989f

10 104 — Schecter et al.

1989f

8.2 56 — Schecter et al.

1989f

32 420 — Schecter et al.

1989f

32 270 — Schecter et al.

1989f

48 330 — Schecter et al.

1989f

7.2 (2.1–17) — Schecter et al. 2,3,7,8-TCDD &

1989g TCDF only

40 255 — Schecter et al.

1990b

42 320 — Schecter et al.

1990b

48 630 — Schecter et al.

1990b

32 156 — Schecter et al.

1990b

32 306 — Schecter et al.

1990b

36 632 — Schecter et al.

1990b
262 T.B. Minh et al.

Table 5. (cont.)

Sample/sampling
Location Year site description Unit n PCDDs

Hochiminh City early Blood pg g−1 lipid 1 (50 1,090

1990s samples
pooled)
Dong Nai, early Blood pg g−1 lipid 1 (33 1,940
southern 1990s samples
Vietnam pooled)
Hanoi early Blood pg g−1 lipid 1 (32 126
1990s samples
pooled)
Hanoi early Breast milk pg g−1 lipid 30 104
1990s
Da Nang, middle early Breast milk pg g−1 lipid 1 (11 406
Vietnam 1990s samples
pooled)
Dong Nai, early Breast milk pg g−1 lipid 1 (11 180
southern 1990s samples
Vietnam pooled)
Hanoi 1987–91 Adipose tissues, pg g−1 lipid 1 (10 270
general samples
population pooled)
Hanoi 1987–91 Adipose tissues, pg g−1 lipid 1 (10 685
soldiers samples
exposed to pooled)
Agent Orange
during the War
Song Be, 1987–91 Adipose tissues, pg g−1 lipid 1 1,050
southern locations in
Vietnam proximity of
Agent Orange
spraying
Hau Giang, 1987–91 Adipose tissues, pg g−1 lipid 1 800
southern locations in
Vietnam proximity of
Agent Orange
spraying
Dong Nai 1987–91 Adipose tissues, pg g−1 lipid 1 226
locations in
proximity of
Agent Orange
spraying
Various locations 1980–91 Whole blood pg g−1 lipid 82 190
from northern (pooled)
Vietnam
Various locations 1980–91 Whole blood pg g−1 lipid 383 860
from southern (pooled)
Vietnam
Various location 1991–92 Blood pg g−1 lipid 82 190
from northern (pooled)
Vietnam
 Organic Pollutants in Vietnam 263

PCDFs Total PCDD/Fs Dioxin-like PCBs Reference Remark

82 1,170 — Schecter et al.

1991

130 2,070 — Schecter et al.

1991

41 167 — Schecter et al.

1991

23 127 — Schecter et al.

1991
130 536 — Schecter et al.
1991

55 235 — Schecter et al.

1991

32 300 — Schecter et al.

1992a

56 740 — Schecter et al.

1992a

68 1,120 — Schecter et al.

1992a

62 860 — Schecter et al.

1992a

23 249 — Schecter et al.

1992a

100 290 — Schecter et al.

1992b

110 970 — Schecter et al.

1992b

100 290 — Schecter et al.

1995
264 T.B. Minh et al.

Table 5. (cont.)

Sample/sampling
Location Year site description Unit n PCDDs

Various location 1991–92 Blood pg g−1 lipid 183 930

from central (pooled)
Vietnam
Various locations 1991–92 Blood pg g−1 lipid 433 760
from southern (pooled)
Vietnam
Hanoi 1999 Whole blood, pg g−1 lipid 1 (100 150
general samples
population pooled)
Bien Hoa, Dong 1999 Whole blood, pg g−1 lipid 20 590 (140–
Nai residents in 1,410)
proximity of
Agent Orange
Air Base
Aso, Aluoi 1999 Whole blood, pg g−1 lipid 78 104
Valley, central males, (pooled)
Vietnam residents in
proximity of
Agent Orange
Air Base
Aso, Aluoi 1999 Whole blood, pg g−1 lipid 85 94
Valley females, (pooled)
residents in
proximity of
Agent Orange
Air Base
Huong Lam, 1999 Whole blood, pg g−1 lipid 64 150
Aluoi Valley males, (pooled)
residents in
proximity of
Agent Orange
Air Base
Huong Lam, 1999 Whole blood, pg g−1 lipid 56 160
Aluoi Valley females, (pooled)
residents in
proximity of
Agent Orange
Air Base
Hong Thuong, 1999 Whole blood, pg g−1 lipid 70 220
Aluoi Valley males, (pooled)
residents in
proximity of
Agent Orange
Air Base
Hong Thuong, 1999 Whole blood, pg g−1 lipid 62 180
Aluoi Valley females, (pooled)
residents in
proximity of
Agent Orange
Air Base
 Organic Pollutants in Vietnam 265

PCDFs Total PCDD/Fs Dioxin-like PCBs Reference Remark

210 1,140 — Schecter et al.

1995

94 850 — Schecter et al.

1995

40 190 29 Schecter et al.

2001

67 (28–184) 660 (190–1,520) 60 (20–156) Schecter et al.

2001

31 135 — Dwernychuk et
al. 2002

33 127 — Dwernychuk et
al. 2002

110 260 — Dwernychuk et

al. 2002

130 290 — Dwernychuk et

al. 2002

84 304 — Dwernychuk et
al. 2002

83 260 — Dwernychuk et
al. 2002
266 T.B. Minh et al.

Table 5. (cont.)

Sample/sampling
Location Year site description Unit n PCDDs

Hong Van, Aluoi 1999 Whole blood, pg g−1 lipid 77 88

Valley males, (pooled)
residents in
proximity of
Agent Orange
Air Base
Hong Van, Aluoi 1999 Whole blood, pg g−1 lipid 64 110
Valley females, (pooled)
residents in
proximity of
Agent Orange
Air Base
Aso, Aluoi 1999 Breast milk, pg g−1 lipid 4 25 (8.8–36)
Valley resident in
proximity of
Agent Organe
Air Base
Huong Lam, 1999 Breast milk, pg g−1 lipid 4 42 (30–68)
Aluoi Valley resident in
proximity of
Agent Organe
Air Base
Hong Thuong, 1999 Breast milk, pg g−1 lipid 4 54 (36–63)
Aluoi Valley resident in
proximity of
Agent Organe
Air Base
Hong Van, Aluoi 1999 Breast milk, pg g−1 lipid 4 34 (19–80)
Valley resident in
proximity of
Agent Organe
Air Base
Tay Mo, suburb 1999 Breast milk, pg g−1 lipid 8 32 (10–81)
Hanoi residents near
dumping site
for municipal
wastes
TEQ (pg g−1 2.7 (1.4–4.8)
lipid)
Tay Mo, suburb 1999 Breast milk, pg g−1 lipid 10 27 (14–41)
Hanoi residents living
far from
dumping site
for municipal
wastes
TEQ (pg g−1 2.9 (1.7–4.0)
lipid)
 Organic Pollutants in Vietnam 267

PCDFs Total PCDD/Fs Dioxin-like PCBs Reference Remark

48 136 — Dwernychuk et
al. 2002

22 130 — Dwernychuk et
al. 2002

7.6 (4.1–16) 33 (13–45) — Dwernychuk et

al. 2002

24 (11–54) 66 (46–92) — Dwernychuk et

al. 2002

32 (11–47) 86 (47–110) — Dwernychuk et

al. 2002

16 (4.4–42) 50 (20–120) — Dwernychuk et

al. 2002

20 (7.3–42) 51 (18–120) 24,000 (4,200– Kunisue et al.

46,000) 2004

3.3 (1.3–7.2) 6.0 (2.9–9.3) 7.5 (1.5–15) Kunisue et al.

2004
20 (9.6–45) 47 (26–67) 15,000 (2,800– Kunisue et al.
22,000) 2004

3.4 (2.0–5.6) 6.3 (3.6–8.1) 5.3 (1.9–13) Kunisue et al.

2004
268 T.B. Minh et al.

In spite of the dioxin problems caused by Agent Orange in southern

Vietnam, as just discussed, there have not been many studies of contamination
status, bioaccumulation characteristics, and the fate and toxic implications
of dioxins in a tropical environment such as Vietnam, and data were not
available until our laboratory reported dioxin contamination in dumping
sites for municipal wastes in 2003 (Minh NH et al. 2003). In recent years,
public media have voiced concern regarding the open dumping sites in
Asian developing countries where large amounts of municipal solid wastes
are dumped. Unfortunately, these open dumping sites are usually located
near human habitats; therefore, exposure to various toxic chemicals from
dumping sites is a matter of serious concern because of the effects on
human health, wildlife, and environmental quality. Natural burning by the
generation of methane gas under anaerobic conditions and combustion by
waste pickers scavenging in dumping sites are favorable factors for the
formation of PCDD/Fs in dumping sites. To fi find an answer to the question
of whether open dumping sites are potential sources of PCDD/Fs,
comprehensive investigations have been conducted to examine PCDD/F
concentrations in soils and human breast milk from open dumping sites
from the Philippines, Cambodia, India, and Vietnam (Minh NH et al. 2003;
Kunisue et al. 2004).
Concentrations of PCDD/Fs in soils from dumping sites in Hanoi were
signifi
ficantly higher than those in Hochiminh. Mean and range concentrations
in dumping sites from Hanoi were 6100 pg/g dry wt [95 pg/g toxicity equiva-
lent (TEQ)] and 125–50,500 pg/g dry wt (0.4–850 pg/g TEQ), respectively
(Minh NH et al. 2003) (Fig. 12). However, in general, dioxin residues in
soils from Vietnam were less than those in other Asian countries examined
in this study, indicating less dioxin contamination in Vietnam. However,
one soil sample contained very high concentration of PCDD/Fs (50 ng/g dry
wt; 850 pg/g TEQ) (Minh NH et al. 2003). This level is greater than most
of the background level soils in industrialized countries and even higher
than those reported for dioxin-contaminated locations in the world (Minh

Fig. 12. Concentrations and TEQs of PCDD/Fs in soils from open dumping sites
for municipal wastes in developing Asian countries (Minh NH et al. 2003).
 Organic Pollutants in Vietnam 269

NH et al. 2003). In addition, PCDD/F levels in soils collected from dumping

sites were significantly
fi higher than those in soils from agricultural and resi-
dential areas far from dumping sites, indicating that open dumping sites can
be a potential source of dioxins in Vietnam. To understand the degree of
human exposure to dioxins in open dumping sites, levels in human milk
samples were also examined (Kunisue et al. 2004). Mean PCDD/Fs
concentration in breast milk from women living near dumping sites in
Hanoi was 52 pg/g lipid (6.0 pg/g TEQs) [range, 18–120 pg/g lipid (2.9–
9.3 pg/g TEQ] (Kunisue et al. 2004). These values are comparable to those
in the Philippines and Cambodia but lower than those in India. In contrast
to soils, there were no significant
fi differences in PCDD/F residues in milk
of women living near dumping sites and far from dumping sites (control
sites) (Kunisue et al. 2004).
On the basis of these results, it can be emphasized that although the
status of contamination in Vietnam was less pronounced as compared to
other countries in the East and South Asian regions, the issue of dioxin
pollution in such open dumping sites should be considered as one of the
priority research targets in the future because of the sporadic evidence of
elevated dioxin contamination in dumping sites from Hanoi. Control
measures and legislation toward management and mitigation of dioxin
emissions in open dumping sites are urgently needed.

IV. Environmental Behavior and Bioaccumulation

A. Transport Behavior in Tropical Environments
Multimedia monitoring studies have not only helped increase knowledge
of the status of contamination but also provided in-depth insights into the
transport behavior and fate of POPs in tropical ecosystems in developing
Asian countries. On a global scale, it has been made clear that the fate of
persistent OCs in the open ocean is different from that in the coastal
environment (Iwata et al. 1993, 1994; Tanabe et al. 1994). The open ocean
plays a role as a fi
final sink for POPs, and cold waters at high latitudes such
as the Arctic Ocean, which are located far from pollution sources, serve as
signifi
ficant reservoirs (Iwata et al. 1993). On the other hand, pronounced
contamination by OC insecticides was observed in coastal waters of Asian
countries as a result of extensive usage for agriculture and public health
purposes (Iwata et al. 1994). On a smaller scale, latitudinal distribution of
persistent OCs such as PCBs, DDTs, and HCHs in sediments from the
southern Vietnamese estuary were also examined (Iwata et al. 1994). The
authors suggested using the relationship between the concentration ratio
of a compound in the water phase (concentration in water sample) and the
particulate phase (organic carbon normalized concentration in sediment)
(S/W ratio) and latitude. The degree of correlation and the ratio (S/W) are
indications of the effi
ficiency of their transport.
270 T.B. Minh et al.

Analytical results showed relatively uniform distribution of HCHs along

the latitudes, and the ratio (S/W) showed the highest degree of positive
correlation with latitude. This phenomenon indicates that HCH isomers
pose higher potency for atmospheric transport and global redistribution.
Other compounds such as DDTs, CHLs, and PCBs with lower correlations
and gradients exhibited less effificiency for atmospheric transport.
In general, the degree of usage, the climate, and the physicochemical
properties of persistent organic compounds are the major factors controlling
the magnitude of contamination. When comparing the distribution of OCs
in different environmental media in the Asia-Pacifi fic region, we found that
distribution patterns in fish and sediment were relatively similar and showed
less spatial variation. The pattern in air and water showed greater geo-
graphic variability. A recent extensive survey of OC insecticides from the
Red and Duong Rivers, the two largest rivers flowing through the northern
delta region of Vietnam, and various lakes in Hanoi has shown that DDT
and HCH levels in rivers is apparently greater than those in lakes (Hung
and Thiemann 2002). This observation suggests different behavior of
semivolatile compounds in tropical lakes and in rivers, showing shorter
retention time in lakes than in rivers. Elevated temperatures may enhance
volatilization of such compounds, leading to shorter residence time in the
tropical water phase, lower levels, and relatively uniform distribution in
sediments and aquatic biota. In this context, the role of the tropics in Asia-
Pacific
fi region as potential emission sources for higher-latitude areas
deserves further attention.
As mentioned earlier, very little information is available regarding
contamination by PCDD/Fs in developing Asian countries. Therefore, the
fate and behavior of these compounds in developing countries are still
obscure. On the basis of residues in soils from dumping sites, fl
flux of PCDD/
Fs to soils can be estimated (Fig. 13). The estimated fl fluxes to soils in
dumping sites in developing Asian countries including Vietnam were sur-
prisingly higher. Fluxes to soils in Asian dumping sites were higher than
those of some other locations in the world, including contaminated areas
in the U.S. and Hong Kong (see Fig. 13). Loads of PCDD/Fs to the dumping
sites were also estimated (Fig. 14). Results indicated that these sites in the
Philippines and India, with a huge area of approximately 23 and 140 ha,
could receive the highest annual amounts of 3900 and 1400 mg/yr PCDD/Fs
(35 and 8.8 mg TEQ/yr), respectively. A dumping site in Hochiminh,
Vietnam, had the lowest loading rate because of less contamination of
PCDD/Fs in soils. For comparison, total annual fluxes
fl to the Kanto region,
Japan, one of the most polluted areas in the world, were found to range
from 50 to 900 g TEQ in a total area of 32,000 km2 (approximately 3 million
ha) (Ogura et al. 2001). The area of dumping sites in India is 140 ha, which
is 21,000 times smaller than that of Kanto region; this area was estimated
to receive 8.8 mg TEQ every year. These data suggest that dumping sites
 Organic Pollutants in Vietnam 271

Philippines
Cambodia
India
Vietnam,, Hanoi
Vietnam,, Hochiminh
Hong Kong
Australia
USA, Indiana
USA, Michigan
Canada, British Columbia
Canada, Yukon
United Kingdom
Spain
Germany
1 10 100 1000 10000 100000
Flux of PCDD/Fs to soils (ng m–2 yr –1)

Fig. 13. Comparison of fluxes to soils from open dumping sites in Asian countries
with those from other locations in the world (Minh NH et al. 2003; Wagrowski and
Hites 2000).

Fig. 14. Comparison of the load of PCDD/Fs to dumping sites in Asian countries
with those in Kanto region, an extensive industrial area in Japan (Minh NH et al.
2003).

in India and the Philippines may be significant

fi reservoirs of PCDD/Fs. Pos-
sible impacts on human health and wildlife living near dumping sites are of
great concern and warrant further comprehensive studies. On the basis of
the results of this study, it is important to note that despite decrease in
global pollution by POPs in the future, developing countries may continue
to be a potential source of certain compounds, particularly PCDD/Fs.
272 T.B. Minh et al.

B. Bioaccumulation in Biota

Surveys of environmental samples and lower trophic level organisms such

as fi
fish and mussels indicate less variable spatial distribution of semivolatile
organic pollutants in sediments and fish on both local and regional scales
(Iwata et al. 1994; Kannan et al. 1995). Despite higher use, HCH levels in
fish from India were lower than those in Japan and the U.S. Similarly, DDT
fi
residues in fishes
fi from India and Vietnam were still lower than those from
the U.S. (Kannan et al. 1995), which indicates rapid volatilization of these
insecticides in the tropical environment, resulting in lower residues in lower
trophic biota. In this context, understanding of bioaccumulation in higher
trophic animals is important to acquire further knowledge on the fate of
persistent OCs in the tropical environment. During autumn and spring
1997, extensive sampling was conducted to collect 101 individual bird
samples comprising 7 species of resident and 17 species of migrant birds
from a wetland on a small island near the Red River estuary, northern
Vietnam (Minh TB et al. 2002). Concentrations of OC insecticides and
PCBs were analyzed in these resident and migratory birds to understand
the magnitude of contamination, bioaccumulation, and metabolism in birds.
Interestingly, an in-depth analysis of the variation of OC residues according
to the feeding habits and migratory behaviors revealed different pictures
of accumulation. In particular, there were no clear differences in OC
accumulation in birds according to their feeding habits. In contrast, the
differences in accumulation patterns according to migratory behavior of
birds were quite apparent. DDTs residues showed higher concentrations in
residents than in migrants, refl flecting the recent usage of DDT in Vietnam,
as we have discussed earlier for other samples. HCH concentrations were
higher in migratory birds as compared to residents, suggesting exposure at
stopover sites in other countries during migration. In this context, the
influence
fl of the rate and degree of OCs local usage may have greater impact
on their accumulation in resident and migratory birds, whereas biological
factors such as feeding habit exhibit less pronounced implications. In
general, results observed in residents and migratory birds from North
Vietnam clearly suggest the suitability of using birds as a bioindicator for
monitoring toxic contaminants.
On the basis of isomer-specifi fic analysis of PCB congeners, it is possible
to estimate the metabolism capacity of contaminants in birds. Tanabe et al.
(1988) suggested the concept of using a “metabolic index” (MI) value to
estimate enzyme activities in higher trophic animals including birds. Higher
MI values imply greater capability to metabolize persistent OCs. Estimated
enzyme activities in some species of resident and migratory birds from
North Vietnam compared with those in other species of birds and mammals
are given in Fig. 15. Estimated PB-type enzyme activities of some species
such as black-capped kingfi fisher and whiskered tern were comparable to
those in the common cormorant but higher than those in other higher-
 Organic Pollutants in Vietnam 273

Fig. 15. Comparison of estimated phenobarbital (PB)- and 3-methylcholanthrene

(MC)-type enzyme activities in higher trophic animals by metabolic indices of
chlorobiphenyl (CB)-52 and CB-66. Black bars represent enzyme activities of
Vietnamese birds (Minh TB et al. 2002).

trophic fish-eating bird species such as kite, puffi

fin, and gull, while MC-type
enzyme activities were comparable or slightly higher in Vietnamese birds.
Higher PB- and, to some extent, MC-type enzyme activities in shorebirds
from Vietnam suggest that these species may have a higher capacity to
metabolize PCB congeners; this may explain relatively lower levels of PCBs
and other OCs in shorebirds from Vietnam compared to high-trophic top
predator species (Minh TB et al. 2002).

V. Temporal Trends
Well-designed studies on temporal trends of POPs contamination in
developing countries are generally limited. This is a common issue in
underdeveloped nations, where advanced knowledge and state-of-the-art
analytical equipment are still substantially lacking. Therefore, reliable and
reproducible data for long-term trends of POP contamination, a key
requirement in monitoring programs, are limited in these countries,
including Vietnam. Nevertheless, studies on temporal trends of contamination
are very important, particularly in tropical developing countries. Few
reports have suggested the role of the southern Asian region as a possible
emission source of toxic contaminants for pristine areas such as the Arctic
274 T.B. Minh et al.

and the Antarctic (Iwata et al. 1993, 1994; Kannan et al. 1995; Kunisue et
al. 2002). Despite the ban on OC usage in most of the developed nations,
high consumption of OC insecticides to enhance food production and
eradicate vector-borne diseases has been a fact in developing countries.
Thus, despite the rapid decrease of OC residues in developed nations, the
status of contamination in the developing world seems different, with slower
rates of decline. Although well-designed studies on the temporal trends of
contamination in POPs from Vietnam have been limited, trends of OC
residues in river water and sediments from the Red River estuary and from
Hochiminh City, and human breast milk from women living in suburb areas
of Hanoi and Hochiminh, were investigated.
Viet (2002) examined concentrations of DDTs and γγ-HCH (lindane), the
two most common insecticides used extensively in Vietnam, in water and
sediments from the Red River delta. River water and sediments were
collected at the same locations annually in both dry and wet seasons
and examined for contamination trends during 1995–2001 (Fig. 16).
DDT residues in water have declined relatively rapidly during 1995–1998
and remained constant until recent years at levels below 10 ng/L.
Concentrations in sediments also exhibited a reducing trend but to a lesser
extent. DDT residues in sediments dropped by a factor of 2 during 1997
and 2000. DDT was officially
fi banned in Vietnam in 1995 (Sinh et al. 1999).
The reduction of DDT concentrations in both water and sediments from
Red River, northern Vietnam, indicate the effect of legislative action to
reduce the degree of pollution. An interesting result was observed on
trends of lindane in sediment, showing peak concentrations in 1997 and
lower levels during 1995–1996 and 1999–2001. Recent studies examined
HCH residues in sediments from different sites in the Red River delta and

max
DDTs
mean
Lindane min

160 Dry season

Concentration (ngg-1 dry wt)
Concentration (ngL-1)

40

120 Water Sediment

30

80 20

Rainy season
40 10

0 0
1995 96 97 98 99 00 01 RS DS RS DS RS DS RS DS RS DS
Year 1995 96 97 99 00 01
Year

Fig. 16. Temporal trends of DDTs and lindane in surface water and sediment from
Red River Delta, northern Vietnam during 1995–2001 (Viet et al. 2002).
 Organic Pollutants in Vietnam 275

estuary and found that HCH concentrations in the 1997 survey were higher
than those analyzed in the 1995 sampling survey (Nhan et al. 1998, 1999).
Such a fluctuating trend of HCH contamination suggests sporadic input of
this insecticide into the Red River watershed. In general, results in water
and sediments in recent years indicated a rapid decline of DDTs and HCHs
in surface water, but a slow reduction in sediment levels.
In southern Vietnam, several studies reported levels of DDTs and PCBs
in urban sediments from Hochiminh metropolitan area (Iwata et al. 1994;
Phuong et al. 1998; Minh NH et al. 2007a). Results of these investigations
in similar sampling areas were compiled to evaluate contamination trends.
PCBs levels decreased from 310 ng/g dry wt in 1990 (Iwata et al. 1994) to
220 ng/g in 1996 (Phuong et al. 1998) and to 82 ng/g in 2004 (Minh et al.
2007a). In the same period, DDTs decreased from 240 ng/g dry wt to 80 and
37 ng/g, respectively.
It is well known that decrease of POP residues in environmental matrices
often follows fi first-order kinetics, and thus their residue level data can be
fitted in simple log-linear regression for examination of temporal trends
fi
(Bignert et al. 1998; Noren and Meironyte 2000). Using this approach, data
from the above studies (Iwata et al. 1994; Phuong et al. 1998; Minh et al.
2007b) were combined to estimate half-life (definedfi as the time needed for
a compound to decrease to one-half of its original concentration) of DDTs
and PCBs in sediments from canals in the metropolitan area of Hochiminh
(Fig. 17). The estimated half-lives were approximately 5 yr for DDTs and
7 yr for PCBs.
The half-life of DDTs in this study is in agreement with those observed
in various environmental matrices from northern Europe (Bignert et al.
1998) as well as those reported for human breast milk in Hochiminh (Minh
et al. 2004a). The estimated half-life for PCBs in sediments from Hochiminh

350
Concentration (ng/g dry wt)

300 PCBs
250 DDTs
200
150
100
50
0
‘89 ‘90 ‘91 ‘92 ‘93 ‘94 ‘95 ‘96 ‘97 ‘98 ‘99 ‘00 ‘01 ‘02 ‘03 ‘04 ‘05
Year of sampling

Fig. 17. Temporal trends of DDTs and PCBs in sediments collected from Hochiminh
City during the last 14 years (Minh NH et al. 2007a).
276 T.B. Minh et al.

was relatively short compared to 11–17 yr reported in the literature (Bignert

et al. 1998). Different methods for quantification
fi of PCBs may have led to
variations in reported total PCBs and thus caused variation in the estima-
tion of PCB half-life. In general, results of previous monitoring studies
suggest there was a somewhat general trend of POP contamination observed
in various ecosystems, i.e., the relatively rapid decline of DDTs and slower
decrease of PCBs (Tanabe et al. 2003). Temporal trends of POP contamina-
tion in the Mekong River delta remain a critical issue that needs long-term
and comprehensive monitoring.
On the other hand, monitoring concentrations in biota should provide
more realistic data to understand trends of environmental contamination
because the impact of any change in the environmental input of persistent
OCs can be realized relatively slowly in biota samples compared to those
observed in environmental abiotic samples. A similar phenomenon was also
seen in the trends of POP pollution in lower and higher trophic animals
(Tanabe et al. 2003).
Minh NH et al. (2004a) assessed the declining rates of human exposure
to DDTs and PCBs over 10 yr (1989 and 2001). A first-order fi kinetics
approach has been used to estimate the declining rates of DDTs and PCBs
in human breast milk from Vietnam (Fig. 18). The decrease of persistent
DDTs, PCBs, and HCHs in human breast milk was suggested to follow
first-order kinetics (Noren and Meironyte 2000). Another important assess-
fi
ment is half-life (tdec1/2). Based on concentrations of OCs in 1989 reported
by Schecter et al. (1989a) and those of 2001 (Minh NH et al. 2004a), the
rate constant and tdec1/2 were estimated.
Levels of p,p′-DDT have decreased from 4700 to 2700 ng/g lipid over a
10-yr period with tdec1/2 of ≈3 yr. On the other hand, p,p′-DDE decreased
rather slowly with a tdec1/2 of 6 yr. These results are somewhat in agreement
with those of Sweden showing half-lives of 4 and 6 yr for p,p′-DDT and
p,p′-DDE (Noren and Meironyte 2000). The slightly shorter half-life
observed here could be because the tropical climate in Vietnam may have
facilitated volatilization of p,p′-DDT in the environment, leading to its
faster decrease in food chains, thus in humans. Given that DDTs exposure
in Vietnam still occurs to some extent, the calculation might have slightly
underestimated half-life values. Nevertheless, the estimated half-life is
fairly useful for assessing the trend of DDT contamination of Vietnamese
breast milk. Assuming that the decrease in the trend of DDTs remains more
or less constant, we can estimate the DDT levels reaching approximately
700 ng/g lipid in the year 2011 (see Fig. 18). However, lower levels in the
future can be expected if the use of DDT is completely phased out. The
decreasing trend of PCBs was lower compared to those of DDT (11–18 yr
for some major congeners such as CB-138, -153, and -180). This result is
somewhat in agreement with those reported from Sweden showing the
half-life for some PCB congeners varying from 11 to 17 yr (Noren and
Meironyte 2000).
 Organic Pollutants in Vietnam 277

8000
7000
p,p’-DDT
Conc. (ng g–1 lipid)
6000
5000
4000
- 0.248x
y = 6250 e
3000
2000
1000
0
1989 1995 2000 2005 2011

8000
7000
p,p’-DDE
Conc. (ng g–1 lipid)

6000
5000
-0.105x
4000 y = 7570 e
3000
2000
1000
0
1989 1995 2000 2005 2011
Year
Fig. 18. Estimation of time-trend curve of p,p′-DDT and p,p′-DDE residues in
human breast milk in Vietnam (Minh NH et al. 2004a).

VI. Environmental and Human Health Implications

Widespread contamination by OC insecticides, particularly DDTs, in
different environmental samples of Vietnam has been apparent, as indicated
in our survey in the early 1990s. In an evaluation of the estuarine sediments
collected from various locations from the northern to southern part of the
country, high concentrations of DDTs were observed (Iwata et al. 1994).
Environment Canada has recently updated the sediment quality guidelines
for protection of aquatic life. The Interim Fresh Water Sediment Quality
Guidelines (ISQGs) and the Probable Effect Levels (PELs) for p,p′-DDE
are 1.42 and 6.75 ng/g dry wt, respectively, whereas these values for p,p′-
DDT are 1.19 and 4.77 ng/g dry wt (Canadian Environmental Quality
Guideline 2002). Among the 18 locations examined throughout Vietnam,
about half of the sediment samples contained p,p′-DDE and p,p′-DDT
greater than the ISQG values. Some samples collected from the municipal
278 T.B. Minh et al.

sewage canal contained elevated levels of DDTs, far exceeding the probable
effect levels (PELs). PCB concentrations in Vietnamese sediments in these
locations were also above the PEL level. Likewise, concentrations of DDTs
in soils collected from some locations from north, middle, and south
Vietnam (Thao et al. 1993a,b) approached or exceeded the guideline
level (700 ng/g dry wt) proposed by Environment Canada and the level of
1000 ng/g dry wt recommended by the Japanese government.
Results of a 2004 survey indicated that concentrations of DDTs in all
sediment samples from Hochiminh canals are higher than the ISQG, and
those in more than half of the samples still exceeded the PEL (Minh TB et
al. 2005; Minh NH et al. 2007a). Nearly half of sediment samples from the
Sai Gon-Dong Nai River had concentrations above the ISQG and 20% of
the samples were above the PEL; only 10% of the estuarine and coastal
samples showed values over the ISQG and no samples over the PEL
(Fig. 19). On the other hand, PCBs levels in all sediments were lower than
the PEL (277 ng/g dry wt; data not shown) and only sediments from Hochim-
inh City canals contained PCBs higher than the ISQG (34 ng/g). The results
demonstrate high toxic potential of sediment in the city canals and there-
fore suggest a need for appropriate management. In this context, it is
important to note that despite the decrease in POPs pollution, present
levels in many sediment samples still exceed the PELs. Input of OCs to the

30

10
Concentration (ng/g dry wt)

9
(A)
8
7
(A)
6
5 (A)
4
(B)
3
2
(B) (B)
1
0

p,p’-DDE p,p’-DDD p,p’-DDT

Hochiminh City canals
Sai Gon – Dong Nai River
Estuary & coastal area

Fig. 19. Concentrations of DDT compounds in sediments from Sai Gon–Dong Nai
River, southern Vietnam in comparison with the Canadian Environmental Quality
Guideline for Sediment (Minh NH et al. 2007a).
 Organic Pollutants in Vietnam 279

aquatic environment is likely the result of the discharge of untreated sewage

from municipal areas, and thus more efficient
fi management on discharge is
needed to prevent discharge of toxic chemicals into the aquatic environment.
Taking into account all these facts, the magnitude of contamination by
DDTs in Vietnam is of concern and warrants further studies. From the
environmental health and global contamination point of view, the role of
Vietnam as a potential source of DDTs for other countries in the region as
well as in higher latitudes should be considered as a priority for future
research.
As for PCDD/Fs, the formation of these contaminants in open dumping
sites in Asian developing countries raises human health concerns, not only
for communities living near the dumping sites but also for people who live
far away, because PCDD/Fs may undergo atmospheric transport and depo-
sition in distant areas. For risk assessment of soils contaminated by dioxins
and related compounds, the Agency for Toxic Substances and Disease
Registry (ATSDR) U.S. Department of Health, proposed guidelines rec-
ommending that areas having soil concentrations within the range 50–
1000 pg TEQ/g should be evaluated for bioavailability, ingestion rates,
community concerns, etc., and that soils with levels >1000 pg TEQs/g should
be considered for stronger actions such as health studies, exposure investi-
gations, etc. (ATSDR guideline, 1997). The Japanese Government recently
issued new standards for dioxins in soil, establishing 1000 pg TEQ/g as the
maximum acceptable level and those within 250–1000 pg TEQ/g be kept
under surveillance. Many soil samples in dumping sites from Asian countries
including Vietnam contained TEQ concentrations exceeding 250 pg/g TEQs
(Fig. 20; Minh NH et al. 2003), suggesting the necessity of continuous moni-
toring. Particularly, some soils from Cambodia and Hanoi dumping sites
contained TEQ concentrations >1000 pg/g, suggesting their potential for
causing adverse health risks for humans and wildlife.
In the perspective of human health implication, surveys conducted in
early 1990s on OCs in foodstuffs provided useful information regarding
dietary intake of these compounds by Vietnamese (Kannan et al. 1992).
The estimated average daily intakes based on exposure through foodstuffs
to PCBs in Vietnam were higher than India and Thailand, comparable to
those reported for developed nations such as the U.S. and Germany.
Particularly, average daily intake of DDTs by Vietnamese was estimated
to be 19 μg/person/d, and this value was the highest of countries in the
region and in developed nations (Kannan et al. 1992). Although the data
used for this estimation were reported a decade earlier, this fact clearly
suggests elevated exposure to DDTs and PCBs by Vietnamese and that the
usage of DDT has been extensive during the past 10 yr.
Surveys in the framework of the recent Asia-Pacific fi Mussel Watch
Program indicated that dietary intakes of DDTs and PCBs from fi fish in
Vietnam were higher than those in Cambodia and Thailand, but still lower
than those in industrialized nations such as Australia, Japan, and Hong
280 T.B. Minh et al.

Fig. 20. Concentrations of PCDD/Fs in soils from dumping sites in developing

Asian countries compared with various environmental guideline values (Minh NH
et al. 2003).

Kong (Monirith et al. 2000). On the basis of data of average seafood

consumption reported by the Food and Agriculture Organization of the
United Nations, the average daily intakes of PCBs and DDTs from seafood
for different countries in the Asia-Pacificfi region were estimated (Table 6).
Interestingly, results again showed that intakes of DDTs by Vietnamese
were apparently higher than those reported in other countries examined.
In addition to the elevated exposure of DDT via seafood to the
Vietnamese general population, certain cohorts living near municipal
dumping sites may be at a higher risk from dioxins and dibenzofurans. A
methodical approach has been developed to evaluate the risk of exposure
to PCDD/Fs via soil ingestion and dermal absorption (Minh NH et al.
2003). Human exposure to PCDD/Fs in soil is considered to be different
for children and adults because of differences in ingestion rates as well as
body weights of children and adults. Intakes of dioxins were estimated to
be the highest in people of the Philippines, followed by Cambodia, India,
Hanoi (North Vietnam), and Hochiminh (South Vietnam). Intakes of
PCDD/Fs by those living near dumping sites in Vietnam were about 2- to
200 fold greater than those of residents in control sites, thus emphasizing
greater health risks. In addition, it is important to note that the estimated
intakes of dioxins via soil ingestion and dermal exposure by children were
higher than those for adults, suggesting greater risk of dioxin exposure for
 Organic Pollutants in Vietnam 281

Table 6. Estimated Daily Intakes of Persistent Organochlorines Via Mussels by Different

Populations in Asia-Pacifi
fic Region.

Seafood Intake of Intake of Intake of

Survey consumptiona PCBsb (ng DDTs (ng HCHs (ng
Country year (g person−1 d−1) person−1 d−1) person−1 d−1) person−1 d−1)

Cambodia 1998 20 15 6.6 <0.2

China 1999–2001 71 180 17,000 57
Hong Kong 1998–1999 69 260 8,300 14
India 1998 13 49 55 26
Indonesia 1998 52 68 52 2.1
Japan 1994 196 5,900 690 63
South Korea 1998 114 420 400 30
Malaysia 1998 156 160 220 <1.6
Philippines 1998 77 440 31 2.3
Russia 1999 54 3,400 650 54
Vietnam 1997 47 66 1,900 2.8
a
Seafood consumption data were cited from FAO Food Balance Sheets, FAO Statistics Divison
(FAO 2006).
b
Intakes were estimated on the basis of residue concentrations in mussels (Asia-Pacific
fi Mussel
Watch Program) reported by Monirith et al. (2003)

g. 21. TEQs concentrations and estimated intakes of PCDD/Fs for breast-fed

children from open dumping sites and control sites in Asian countries and Aluoi
Valley, a former Agent Orange spraying target during the Vietnam War (Kunisue
et al. 2004; Dwernychuk et al. 2002).

children near dumping sites (Minh NH et al., 2003). Further investigations

should be focused on children and infants, as they are the most susceptible
group and have greater exposure to dioxins. Breast-fed children intakes of
PCDD/Fs estimated on the basis of residues in breast milk of women living
near open dumping sites in Asian countries are given in Fig. 21. The intakes
estimated for Vietnamese were comparable to those in Cambodia but lower
282 T.B. Minh et al.

than in the Philippines and India. In addition, intakes estimated for children
living near the hot spot of dioxin contamination from Agent Orange in
southern Vietnam are still very high, even after spraying ended almost
three decades ago (see Fig. 21; Dwernychuk et al. 2002). Thus, Vietnam
could serve as a suitable location for future research on possible toxic
effects of dioxins on wildlife and humans as this is a place where a unique
situation of both current and historical dioxin contamination exists.

VII. Conclusions and Recommendations

Multimedia monitoring studies have made substantial contributions to the
discovery, description, and understanding of the environmental problems
caused by persistent man-made chemicals. Over the past two decades,
comprehensive surveys were conducted in the Asia-Pacific fi region, including
Vietnam, to make clear various issues of POPs contamination such as
extent of pollution, transport behavior and fate, spatial and temporal trends,
and toxic implications for the environment, wildlife, and human health. The
following conclusions can be made on the basis of these monitoring
studies.

• POPs contamination is still widespread in various environmental media

in Vietnam. In particular, DDT, a well-known insecticide that is economic
and effective for both agricultural crop production and eradication of
vector-borne diseases in developing countries, is the prime contaminant
in Vietnam. Elevated DDT levels found in various environmental media
such as water, soils, sediment, fi
fish, bivalves, and birds in both past and
recent monitoring surveys indicate that Vietnam may be a potential
source of DDT in the South Asia region. More importantly, this insec-
ticide poses high bioaccumulation potential in the food chain and
eventually bioaccumulates in the human body to a substantial extent.
Relatively high levels of DDTs found in human breast milk from both
Hanoi and Hochiminh warrant relatively higher risk for breast-fed
children.
• In general, the hot spots of POPs pollution were linked closely with
extensive industrial and human activities. Elevated levels were observed
in the metropolitan areas in Hanoi and Hochiminh as well as urban sites
along the Mekong River. This fact suggests that the fate of persistent
organic pollutants such as DDTs, HCHs, PCBs, and PBDEs in the
Vietnamese environment seems to be governed by human activities
rather than their physicochemical properties such as “long-range
atmospheric transport potency.” In addition, our results clearly
demonstrate that open dumping sites in developing Asian countries,
including Vietnam, are new potential hot spots of pollution by various
toxic chemicals, particularly dioxins and DDTs.
 Organic Pollutants in Vietnam 283

• Several studies have been conducted to understand the levels and impacts
of dioxins in specifific hot spots from southern Vietnam as a result of Agent
Orange spraying during the war. Surveys conducted during the late 1980s/
early 1990s indicate elevated levels of dioxins from these sites. In addition,
our recent studies have raised a concern regarding dioxin contamination
in open dumping sites in developing Asian countries, suggesting their
role as a potential source of dioxins and related compounds. Although
dioxin residues in soils and human milk collected from dumping sites in
Vietnam were generally lower than those in other Asian countries, the
status of dioxin pollution in these dumping sites was much greater than
those in control sites. The role of dumping sites as a source of dioxins,
therefore, is an important environmental issue for future research.
• For temporal trends of POPs pollution, available data demonstrate a
relatively rapid decline in OC insecticide residues such as DDTs and
HCHs in the Vietnamese environment, while PCBs show slower
decreasing trends. In this context, it is important to note that despite the
fact that DDT residues have declined over the last decade, the present
levels are still high and well exceed the environmental quality guideline
values.
• Open dumping sites for municipal wastes in developing Asian countries
are potential sources of many chemical contaminants. Long-term health
impacts of POPs on humans living in and around these dumping sites are
probably a major concern for future research in Vietnam.
• With rapid growth of industrial activities, the use of modern chemicals
such as PBDEs as flame retardants is another issue that needs particular
attention. In this context, the role of “e-waste,” the specifi
fic dumping sites
of used electronic devices and appliances and computers, as a significantfi
source of modern and toxic chemicals such as polybrominated com-
pounds, should be elucidated in developing Asian countries including
Vietnam.
• Considering these critical issues of POPs pollution, it is clear that con-
tinued and constant efforts should be made to deal with environmental
problems. Despite the possible decrease in global contamination by POPs
in the future, developing countries in the Asia-Pacific
fi region may continue
to be a potential sources for certain contaminants such as DDTs and
PCDD/Fs. Comprehensive and long-term monitoring programs are still
urgently needed. In this context, well-designed monitoring of temporal
trends of POP residues in developing countries, including Vietnam, over
an extended period is crucial for tracing the unrevealed sources and
predicting future prospects of their pollution. In addition, the issue of
toxic chemicals exposure to humans, particularly those currently residing
in extremely poor living conditions, such as near open dumping sites for
municipal wastes in developing countries, should be the major strategy
for future research.
284 T.B. Minh et al.

Summary
This review provides a comprehensive overview of the contamination by
persistent organic pollutants (POPs) in Vietnam based on the results of
extensive monitoring studies conducted during the last two decades.
Available data on POP levels in various environmental media and humans
are compiled and discussed to help provide in-depth insights into their
distributions and sources, transport behavior and fate, bioaccumulation
features, temporal trends, and the potential impacts on ecosystems and
human health. Surveys conducted in the framework of the Asia-Pacific fi
Mussel Watch Program during early 1990s indicated widespread
contamination by PCBs and organochlorine insecticides, particularly DDT
and HCH in various environmental compartments such as air, water, soils,
sediments, and fish collected from different parts of Vietnam. Recent
studies continued to reveal elevated contamination by DDTs in fi fish, mussels,
and birds from Vietnam. DDT concentrations in fish and birds from Vietnam
are among the highest reported for countries in the Asia-Pacifi fic region,
suggesting the role of Vietnamese environment as a potential emission
source of DDTs in this region. Although residues of this insecticide in
Vietnam have declined relatively rapidly during the past decade, recent
levels still exceed the proposed guideline values proposed by Canada and
the U.S. Open dumping sites for municipal wastes in some major cities
such as Hanoi and Hochiminh are a matter of concern with regard to
environmental pollution, particularly contamination by DDTs and PCDD/
Fs. Daily intakes of DDTs via seafood estimated for the Vietnamese general
population were among the highest reported for East Asian countries. In
the open dumping sites, intakes of dioxins by residents were signifi ficantly
greater than those living far from dumping sites. Particularly, the estimated
intakes of dioxins via soil ingestion and dermal exposure for children were
higher than those for adults, suggesting greater risk of dioxin exposure for
children near dumping sites. Future studies should be focused on the tem-
poral trends of POPs in biota in Vietnam to predict the future trends of
contamination and to understand possible toxic impacts on organisms. In
addition, human exposure and possible toxic effects, particularly on chil-
dren, should be considered as priority research, because they are the most
susceptible group and have greater exposure to POPs.

Acknowledgments
We thank Annamalai Subramanian, CMES, Ehime University, Japan for
his critical reading of this manuscript. This study was supported by grants
from the Environmental Science and Technology in the Core University
Program between Japan Society for the Promotion of Science (JSPS) and
National Center for Natural Science and Technology, Vietnam (NCST),
Research Revolution 2002 (RR2002) of Project for Sustainable Coexistence
 Organic Pollutants in Vietnam 285

of Human, Nature and the Earth (FY2002) and “21st Century Center of
Excellence (COE) Program” from the Ministry of Education, Culture,
Sports, Science and Technology, Japan.

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Manuscript received April 25; accepted May 1, 2007.

 Index

Abbreviations used in carbamate Aldicarb, metabolic pathway

QSAR, 208 physiological model (illus.), 207
Acetolactate synthase (ALS) Aldicarb/metabolites, water solubility
inhibition, triazolopyrimidine (illus.), 77
herbicides, 31 Aldicarb toxicity, 61, 62
Acetylcholinesterase (AChE), 55 ALS (acetolactate synthase) inhibition,
AChE (acetylcholinesterase), 55 triazolopyrimidine herbicides, 31
AChE available for inhibition of ALS inhibitors, herbicide families, 31
carbamates (table), 105 ALS-inhibiting herbicides, weed
AChE, binding sites (illus.), 99 resistance, 31, 37
AChE, carbamate inhibition kinetic Analytical methods, triazolopyrimidine
constants, 115 herbicides, 38
AChE, determination in crude tissue Aniline ring para-hydroxylation,
preparations, 103 triazolopyrimidine herbicides
AChE, kinetic activity, 111 metabolism, plants, 45
AChE, molecular isoforms (table), 100
AChE, purification
fi methods, 108 ß-esterase, active sites vs tissue
AChE, structure, multiple forms, 98 concentrations, 104
Acronyms used in carbamate QSAR, ß-esterases, described, 55
208 Bagasse fl fly ash, contaminated
ACSL (Advanced Continuous groundwater remediation, 15
Simulation Language), 55 Bagasse pith, dye-contaminated
Activated carbon, contaminated groundwater remediation, 15
groundwater remediation, 13 BChE (butyrylcholinesterase, 55
Activated sludge, contaminated BChE, carbamate inhibition kinetic
groundwater remediation, 14 constants, 116
Acylation of ChEs and CaEs, 109 BChE, structure, multiple forms, 101
Adipose : Blood partition coefficients,
fi Bioaccumulation, organochlorines
carbamates (illus.), 76 biota, Vietnam, 272
Adipose : Plasma partition coefficients,
fi Biological parameters required for
carbamates (illus.), 75 PBPK/PD models (table), 55
Advanced Continuous Simulation Biological parameters used in PBPK/
Language (ACSL), 55 PD models, 68
Agent Orange, 2,4,5-T, Vietnam, 215, Bioremediation, organochlorine-
244 contaminated soils, 11
Agent Orange, TCDD contaminant, Bioventing, organochlorine-
215, 244 contaminated soils, 12
Agent Orange, Vietnam war, 214 Birds, organochlorines, Vietnam
Aldicarb, ChEs inhibition, 116 (illus.), 242
Aldicarb, in vivo metabolism model, Blood, organochlorines, Vietnam
87, 95 (table), 244 ff.

291
292 Index

Bolus dose, carbamate QSAR, 68 Carbaryl, biotransformation &

Breast milk, organochlorines, Vietnam elimination paths, 189
(table), 244 ff. Carbaryl, ChEs inhibition, 116
Butyrylcholinesterase (BChE), 55 Carbaryl, in vivo metabolism model,
88, 95
CaE (carboxylesterase), 55 Carbaryl, liver microsomal values, 205
CaE, structure, multiple forms, 102 Carbaryl/metabolites, water solubility
CaEs, carbamylation & (illus.), 78
decarbamylation, 112 Carbaryl toxicity, 61, 62
Carbamate bioavailability classification,
fi Carbofuran, biotransformation &
64 elimination paths, 193
Carbamate hydrolysis, via carboxyl Carbofuran, ChEs inhibition, 117
esterases, 86 Carbofuran, in vivo metabolism model,
Carbamate in vitro metabolism, liver 89, 96
microsomes, 84 Carbofuran, liver microsomal values,
Carbamate insecticides, general 205
description, 56 Carbofuran toxicity, 61, 62
Carbamate metabolic pathways, 187 ff. Carboxylesterase (CaE), 55
Carbamate metabolism, CAS numbers, triazolopyrimidine
glucuronidation & transcellular herbicides, 33
transport, 82 CASE, QSAR model software
Carbamate metabolite chemical package, 65
structures, 142 ff. CE-UV, herbicide analytical methods,
Carbamate metabolites metabolism 38
pathways, 187 ff. Chemical expressions (used in
Carbamate pesticide human risk carbamate QSARs), 210
assessment, 53 ff. Chemical names, triazolopyrimidine
Carbamate pesticide PBPK/PD herbicides, 33
Models, human risk assessment, Chemical structures, carbamate
53 ff. metabolites, 142 ff.
Carbamate pesticide QSAR Models, Chemical structures, carbamates,
human risk assessment, 53 ff. 142 ff.
Carbamate pesticide toxicity, 60 ff. Chemical structures, triazolopyrimidine
Carbamate toxicity models, 64 herbicides, 35
Carbamates, acute dermal toxicity, 61 ChEs, acylation, 109
Carbamates, acute oral toxicity, 60 ChEs, carbamylation &
Carbamates, chemical structures, decarbamylation, 112
142 ff. ChEs, decarbamylation, 109
Carbamates, gastrointestinal Chile, organochlorine-contamination
absorpbion, 68 remediation, 1 ff.
Carbamates, human oral Chile, organochlorine-contaminated
bioavailability, 63 sites, 2
Carbamates, multiple ring, described, Chlordane, Vietnam, 215
58 Chlordane, Vietnam sediments/soils
Carbamates, N-acylated, described, 59 (table), 218 ff.
Carbamates, oxime, described, 59 Chlorinated hydrocarbon industrial
Carbamates, physical parameters, 142 ff. solvents, 4
Carbamates, tissue : partition Chlorinated hydrocarbons, Chilean
coeffificients, 142 ff. contaminated sites, 3
 Index 293

Chloroform, industrial contaminant, 4 Developing countries, organochlorine-

Chlorophenols, Chilean contaminated contamination, remediation,
sites, 3 1 ff.
Chlorophenols, groundwater Dibenzofurans (PCDD/Fs), Vietnam,
contaminants, 4 215
Chlorophenols, physicochemical Dichloromethane (DCM), industrial
properties (table), 10 contaminant, 4
Chlorophenols, regulated by EPA Diclosculam, physicochemical
Cluster Rule, 5 properties, 33
Chlorophenols, vadose zone Dimethylcarbamates, fifirst synthesized
contamination, 11 carbamate insecticides, 57
Cloransulam-methyl, physicochemical Dioxin contamination, Vietnam soils/
properties, 33 sediments, 244
Cluster Rule, chlorophenols, EPA Dioxins (polychlorinated dibenzo-p
- -
regulations, 4 dioxins), Vietnam, 215
COMPACT, QSAR model software Dose-response pharmacokinetics,
package, 65 carbamate QSAR, 69
Compost limit values, organochlorine
contaminants, 20 ELISA, herbicide analytical methods,
Contaminant limit values, 38
organochlorine contaminants, Environmental contamination by
landfifill type (table), 19 POPs, Vietnam, 213 ff.
Corn cobs, contaminated groundwater Environmental fate/occurrence,
remediation, 15 triazolopyrimidine herbicides, 39
Cytochrome P450 enzyme, described, Enzyme activities, rat tissues (table),
83 104
Cytochrome P450, multiple forms, 84 European Union, organochlorine-
contaminated sites, 2, 3
DCM (dichloromethane), industrial
contaminant, 4 Fish, organochlorines levels, Vietnam
DDT, human health implications, (table), 232, 240
Vietnam, 277 Florasulam, photodegradation pathway
DDT, malaria control, amount used, (diag.), 42
Vietnam, 215, 216, 228 Florasulam, physicochemical
DDT soil levels, countries compared, properties, 33
231 Flumetsulam, chemical synthesis
DDT, temporal trends, sediments, scheme (diag.), 36
Vietnam, 275 Flumetsulam metabolism, tolerant
DDT, Vietnam, 215 corn, wheat, barley (diag.), 47
DDT, Vietnam sediments/soils (table), Flumetsulam, physicochemical
218 ff. properties, 33
Decarbamylation of ChEs and CaEs, Fly ash, contaminated groundwater
109 remediation, 14
Decontaminating methods, Formetanate, biotransformation &
organochlorines, 1 ff. elimination paths, 195
DEREK, QSAR model software Formetanate, ChEs inhibition, 117
package, 65 Formetanate, in vivo metabolism
Developing countries, organochlorine model, 90, 96
contamination, 1 ff. Formetanate toxicity, 61, 62
294 Index

Gastrointestinal absorption, Mathematical expressions, used in

carbamates, 68 carbamate QSARs, 210
GC-MS, herbicide analytical methods, Mekong River Delta, Vietnam,
38 contaminants, 214
Glucuronidation & transcellular Methiocarb, biotransformation &
transport, carbamate metabolism, elimination paths, 196
82 Methiocarb, ChEs inhibition, 117
Greenfifields, defi
fined, 2 Methiocarb, in vivo metabolism model,
Groundwater remediation, 90, 96
organochlorine contamination, 13 Methiocarb toxicity, 61, 62
Methomyl, biotransformation &
Halogen-substituted phenyl-, thio-, elimination paths, 197
nitrophenylcarbamates, Methomyl, ChEs inhibition, 118
descriptions, 58 Methomyl, in vivo metabolism model,
Hanoi, Vietnam, organochlorine 91, 96
contaminants, 229 Methomyl toxicity, 61, 62
HCH (hexachlorocyclohexane) Metosulam, physicochemical
isomers, Vietnam, 215 properties, 33
HCHs, Vietnam sediments/soils Mode of action, triazolopyrimidine
(table), 218 ff. herbicides, 31
Heavy metals, contaminated MULTICASE, QSAR model software
groundwater remediation, 15 package, 65
Herbicides, sulfonamides, fate/ Multiple ring carbamates, described, 58
chemistry, 31 ff. Mussel contamination, PCBs, Chile, 8
Hexachlorocyclohexane (HCH) Mussels, organochlorines
isomers, Vietnam, 215 contamination, Vietnam (table),
Hochiminh City, Vietnam, 232
organochlorine contaminants, 229
Human adipose, organochlorines, N-acylated carbamates, described, 59
Vietnam (table), 244 ff.
Human exposure to POPs, Vietnam, ONCOLOGIC, QSAR model software
213 ff. package, 65
Human risk assessment, carbamate Organochlorine compounds, treatable
pesticides, 53 ff. with Fe0 reactive wall (table), 16
Organochlorine soil levels, countries
Imidazolinone herbicides, ALS compared, 231
inhibitors, 31 Organochlorine-contaminated sites,
Insecticide-contaminated sites, European Union, 2, 3
remediation methods, 1 ff. Organochlorine-contaminated sites,
remediation methods, 1 ff.
LC-MS/MS, herbicide analytical Organochlorine-contaminated soils,
methods, 38 remediation (diag.), 18
LC-UV, herbicide analytical methods, Organochlorines, bioaccumulation,
38 aquatic mammals, Vietnam, 273
Lindane, temporal trends, water/ Organochlorines, bioaccumulation,
sediments, Vietnam, 274 birds, Vietnam, 273
Liver microsomal P450 & CYP Organochlorines, bioaccumulation,
hydroxylation models, 66 terrestrial animals, Vietnam, 273
 Index 295

Organochlorines, bioaccumulation, PBPK/PD (physiologically based

tropics, Vietnam, 269 pharmacokinetic/
Organochlorines, biota pharmacodynamic) models, 54
bioaccumulation, Vietnam, 272 PBPK/PD models, biological
Organochlorines, daily intake paramaters required (table), 55
(mussels), Asia-Pacific fi region PCB reductive dechlorination, using
(table), 281 zero-valent iron , 15
Organochlorines, defined,
fi 3 PCB usage, amount, Vietnam, 215
Organochlorines, environmental PCBs, amounts in Chilean regions
behavior, tropics (Vietnam), 269 (table), 7
Organochlorines, human exposure, PCBs, Chilean contaminated sites, 3
Vietnam, 243 PCBs, global production, 5
Organochlorines, human health PCBs, mussels contamination, Chile, 8
implications, Vietnam, 277 PCBs, physicochemical properties
Organochlorines, human samples, (table), 6
Vietnam (table), 238, 244 ff. PCBs soil levels, countries compared,
Organochlorines, transport behavior, 231
tropics (Vietnam), 269 PCBs, temporal trends, sediments,
Organochlorines, Vietnam biological Vietnam, 275
samples (table), 232, 244 ff. PCBs, Vietnam, 215
Organochlorines, Vietnam birds PCBs, Vietnam biological samples
(illus.), 242, 273 (table), 244 ff.
Organochlorines, Vietnam fi fish (table), PCBs, Vietnam, breast milk (table),
232, 240 244 ff.
Organochlorines, Vietnam foods, 242 PCBs, Vietnam, human adipose
Organochlorines, Vietnam Hau River (table), 244 ff.
(map), 228 PCBs, Vietnam sediments/soils (table),
Organochlorines, Vietnam mussels 218 ff.
(table), 232 PCBs, Vietnam soils/sediments (table),
Organochlorines, Vietnam sediments/ 244 ff.
soils (table), 218 ff. PCDD/Fs (dibenzofurans), Vietnam,
Organochlorines, Vietnam water 215
(table), 217 PCDD/Fs, Vietnam biological samples
Oxamyl, biotransformation & (table), 244 ff.
elimination paths, 199 PCDD/Fs, Vietnam, breast milk
Oxamyl, ChEs inhibition, 118 (table), 244 ff.
Oxamyl, in vivo metabolism model, 92, PCDD/Fs, Vietnam, human adipose
96 (table), 244 ff.
Oxamyl toxicity, 61, 62 PCDD/Fs, Vietnam soils/sediments
Oxime carbamates, described, 59 (table), 244 ff.
PCE (perchloroethylene), industrial
P450 & CYP hydroxylation models, 66 contaminant, 4
P450, cytochrome enzyme, described, Penoxsulam, anaerobic metabolism,
83 flooded soils (diag.), 46
fl
Partition coeffi
ficients spreadsheet, Penoxsulam, phtodegradation
aldicarb acid, 80 pathways, 41, 43
PBDEs (polybrominated biphenyl Penoxsulam, physicochemical
ethers), Vietnam, 215 properties, 33
296 Index

Perchloroethylene (PCE), industrial Pirimicarb toxicity, 61, 62

contaminant, 4 Polybrominated biphenyl ethers
Persistent organic pollutants (POPs), (PBDEs), Vietnam, 215
integrated remediation, 16 Polychlorinated biphenyls, Chilean
Persistent Organic Pollutants (POPs), contaminated sites, 3
See also Organochlorines, 217 Polychlorinated dibenzo-p - -dioxins
Persistent Organic Pollutants (POPs), (dioxins), Vietnam, 215
See also Persistent POPs (Persistent organic pollutants),
Organochlorines, 217 integrated remediation, 16
Persistent Organic Pollutants in POPs (Persistent Organic Pollutants)
Vietnam (POPs), 213 ff. in Vietnam, 213 ff.
Persistent organochlorines, Vietnam POPs, soil washing remediation, 17
biological samples (table), 232 ff. POPs, wastewater treatment
Persistent organochlorines, Vietnam remediation, 17
human samples (table), 238 POPs, windrow composting
Persistent organochlorines, Vietnam remediation, 20
sediments/soils (table), 218 ff. Propoxur, biotransformation &
Persistent organochlorines, Vietnam elimination paths, 200
water (table), 217 Propoxur, ChEs inhibition, 118
Persistent Organochlorines, See also Propoxur, in vivo metabolism model,
Persistent Organic Pollutants, 213 93, 97
Persistent Organochlorines, See also Propoxur toxicity, 61, 62
Organochlorines, 217 Pump and treat, organochlorine-
Pesticide-contaminated sites, contaminated groundwater, 13,
remediation methods, 1 ff. 15
Pesticides, carbamates, human risk Purification
fi of enzymes, 108
assessment, 53 ff. Pyrimidinyl thiobenzoate herbicides,
Phenylmethylcarbamates, early ALS inhibitors, 31
investigations, 57 Pyroxsulam, physicochemical
Photodegradation pathway, florasulam
fl properties, 33
(diag.), 42
Photodegradation, triazolopyrimidine QSAR (Quantitative structure-activity
herbicides, 40 relationship), 54
Physical parameters, carbamates, QSAR models for predicting model
142 ff. biological parameters, 62
Physicochemical properties, PCBs QSPBPK/PD (Quantitative structure
(table), 6 physiologically based
Physicochemical properties, pharmacokinetic/dynamic models),
triazolopyrimidine herbicides, 32 54
Physiologically based pharmacokinetic/ Quantitative structure-activity
pharmacodynamic (PBPK/PD) relationship (QSAR), 54
models, 54
Physostigmine, first
fi known carbamate, Rate of ingestion, carbamate QSAR,
57 68
Pirimicarb, biotransformation & Red River Delta, Vietnam, 214
elimination paths, 200 Remediation methods, organochlorine-
Pirimicarb, ChEs inhibition, 118 contaminated sites, 1 ff.
Pirimicarb, in vivo metabolism model, Remediation, organochlorine-
92, 97 contaminated soils (diag.), 18
 Index 297

Remediation techniques, Thiodicarb, ChEs, inhibition, 118

organochlorine-contaminated sites, Thiodicarb, in vivo metabolism model,
8 93, 97
Thiodicarb toxicity, 61, 62
Sample filtration
fi schematic, carbamates Tissue : Blood partition coefficients,
fi
QSAR, 72 carbamates, 66, 73
Sawdust, contaminted groundwater Tissue partition coefficients,
fi
remediation, 15 carbamates, 142 ff.
Skin absorption, carbamate QSAR, 68 Tissue partition coefficients/
fi
Skin : Air partition coefficients,
fi distribution, carbamates, 71
carbamates, 69 TOPKAT, QSAR model software
Skin : Blood partition coefficients,
fi package, 54, 65
carbamates, 69 Toxicity, carbamate pesticides, 60 ff.
Skin : Vehicle partition coefficients,
fi Toxicity equivalent (TEQ), 268, 281
carbamates, 69 Toxicological properties,
Soil remediation, organochlorine triazolopyrimidine herbicides, 33
contamination, 11 Trade names, triazolopyrimidine
Soil washing, organochlorine- herbicides, 33
contaminated soils, 12, 17 Triazolopyrimidine herbicides, aerobic
Soil washing remediation, POPs, 17 soil metabolism, 44
Soils, contaminated groundwater Triazolopyrimidine herbicides,
remediation, 14 analytical methods, 38
Solfonamide herbicides, fate/chemistry, Triazolopyrimidine herbicides, animal
31 ff. metabolism, 43
South America, organochlorine- Triazolopyrimidine herbicides, CAS
contamination remediation, 1 ff. numbers, 33
Substrate selectivities, AChE, BChE, Triazolopyrimidine herbicides,
CaE, 102 chemical structures, 35
Sugar beet pulp, contaminated Triazolopyrimidine herbicides,
groundwater remedication, 15 environmental fate/occurrence, 39
Sugar cane bagasse, contaminated Triazolopyrimidine herbicides, fate/
groundwater remediation, 15 chemistry, 31 ff.
Sulfonylurea herbicides, ALS Triazolopyrimidine herbicides,
inhibitors, 31 microorganism metabolism, 43
Triazolopyrimidine herbicides,
T 2,4,5-T, Agent Orange, Vietnam, 215 photodegradation, 40
TCA (trichloroethane) industrial Triazolopyrimidine herbicides,
contaminant, 4 physicochemical properties, 32
TCDD (tetrachlorodibenzo-p - -dioxin), Triazolopyrimidine herbicides, plant
Agent Orange, Vietnam, 215, 244 metabolism, 43
TCE (trichloroethylene), industrial Triazolopyrimidine herbicides,
contaminant, 4 toxicological properties, 33
Temporal trends, organochlorines Triazolopyrimidine herbicides, trade
environment, Vietnam, 274 names, 33
TEQ (toxicity equivalent), 268, 281 Triazolopyrimidine sulfonamide (TSA)
Tetrachlorodibenzo-p- -dioxin (TCDD), herbicides, fate/chemistry,
Agent Orange, Vietnam, 215, 244 31 ff.
Thiodicarb, biotransformation & Triazolopyrimidine sulfonamide
elimination paths, 203 herbicides, mode of action, 31
298 Index

Trichloroethane (TCA), industrial Wastewater limit values,

contaminant, 4 organochlorine contaminants,
Trichloroethylene (TCE), industrial 19
contaminant, 4 Wastewater treatment remediation,
TSP-LC-MS, herbicide analytical POPs, 17
methods, 38 Water solubility, aldicarb/metabolites
(illus.), 77
UNEP (United Nations Environment Water solubility, carbaryl/metabolites
Program), Vietnam, 215 (illus.), 78
Weed resistance, ALS-inhibiting
Vietnam, Persistent Organic Pollutants herbicides, 31, 37
(POPs), 213 ff. Windrow composting remediation,
Vietnamese exposure to POPs, 213 ff. POPs, 20