ACETONE

EPA/635/R-03/004
www.epa.gov/iris
TOXICOLOGICAL REVIEW
OF
ACETONE
(CAS No. 67-64-1)
In Support of Summary Information on the Integrated Risk

Information System (IRIS)
May 2003
U.S. Environmental Protection Agency

Washington, DC
DISCLAIMER
This document has been reviewed in accordance with U.S. Environmental Protection
Agency policy and approved for publication. Mention of trade names or commercial products
does not constitute endorsement or recommendation for use. Note: This document may undergo
revisions in the future. The most up-to-date version will be made available electronically via the
IRIS Home Page at http://www.epa.gov/iris.
ii
CONTENTS—TOXICOLOGICAL REVIEW OF ACETONE (CAS No. 67-64-1)
FOREWORD . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v
AUTHORS, CONTRIBUTORS, AND REVIEWERS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vi
1. INTRODUCTION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
2. CHEMICAL AND PHYSICAL INFORMATION
RELEVANT TO ASSESSMENTS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
3. TOXICOKINETICS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
3.1. ABSORPTION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
3.1.1. Oral Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
3.1.2. Inhalation Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
3.1.3. Dermal Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
3.2. DISTRIBUTION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
3.3. METABOLISM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
3.4. EXCRETION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
3.5. PBPK MODELING . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
4. HAZARD IDENTIFICATION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
4.1. STUDIES IN HUMANS — EPIDEMIOLOGY AND CASE REPORTS . . . . . . . 22
4.1.1. Cancer Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
4.1.2. Noncancer Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
4.1.2.1. Controlled Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
4.1.2.2. Cohort Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24
4.1.2.3. Case Reports . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
4.2. PRECHRONIC AND CHRONIC STUDIES AND CANCER
BIOASSAYS IN ANIMALS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28
4.2.1. Prechronic Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28
4.2.1.1. Oral Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28
4.2.1.2. Inhalation Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
4.2.2. Chronic Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32
4.2.3. Cancer Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32
4.3. REPRODUCTIVE AND DEVELOPMENTAL STUDIES . . . . . . . . . . . . . . . . . . 32
4.3.1. Reproductive Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32
4.3.1.1. Oral Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32
4.3.2. Developmental Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33
4.3.2.1. Oral Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33
4.4. OTHER STUDIES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
4.4.1. Neurotoxicity Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
4.4.1.1. Oral Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
iii
4.4.2. Isopropanol Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39
4.4.2.1. Oral Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39
4.4.3. Genotoxicity Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
4.5. SYNTHESIS AND EVALUATION OF MAJOR NONCANCER EFFECTS
AND MODE OF ACTION — ORAL AND INHALATION . . . . . . . . . . . . . . . 45
4.6. WEIGHT-OF-EVIDENCE EVALUATION AND CANCER
CHARACTERIZATION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49
4.7. SUSCEPTIBLE POPULATIONS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50
4.7.1. Possible Childhood Susceptibility . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50
4.7.2. Possible Gender Differences . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51
4.7.3. Other Possible Susceptible Populations . . . . . . . . . . . . . . . . . . . . . . . . . . 51
5. DOSE-RESPONSE ASSESSMENTS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52
5.1. ORAL REFERENCE DOSE (RfD) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52
5.1.1. Choice of Principal Study and Critical Effect . . . . . . . . . . . . . . . . . . . . . 52
5.1.2. Methods of Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54
5.1.3. RfD Derivation - Including Application of Uncertainty Factors (UFs) . . 55
5.2. INHALATION REFERENCE CONCENTRATION (RfC) . . . . . . . . . . . . . . . . . . 57

5.3. CANCER ASSESSMENT . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60
6. MAJOR CONCLUSIONS IN THE CHARACTERIZATION
OF HAZARD AND DOSE RESPONSE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60

6.1. HAZARD POTENTIAL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60
6.2. DOSE RESPONSE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62
7. REFERENCES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62
APPENDIX A. SUMMARY OF EXTERNAL PEER REVIEW
AND PUBLIC COMMENTS AND DISPOSITION . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72
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FOREWORD
The purpose of this Toxicological Review is to provide scientific support and rationale
for the hazard and dose-response assessment in IRIS pertaining to chronic exposure to acetone.
It is not intended to be a comprehensive treatise on the chemical or toxicological nature of
acetone.
In Section 6, EPA has characterized its overall confidence in the quantitative and
qualitative aspects of hazard and dose response. Matters considered in this characterization
include knowledge gaps, uncertainties, quality of data, and scientific controversies. This
characterization is presented in an effort to make apparent the limitations of the assessment and
to aid and guide the risk assessor in the ensuing steps of the risk assessment process.
For other general information about this assessment or other questions relating to IRIS,
the reader is referred to EPA’s IRIS Hotline at 202-566-1676.
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AUTHORS, CONTRIBUTORS, AND REVIEWERS
CHEMICAL MANAGERS
Lynn Flowers, Ph.D., DABT
National Center for Environmental Assessment
Washington, DC
Michael W. Broder, Ph.D.
Washington, DC
AUTHORS
Lynn Flowers, Ph.D., DABT
Washington, DC
Michael W. Broder, Ph.D.
Washington, DC
Carol Forsyth, Ph.D.
Oak Ridge National Laboratory
Oak Ridge, TN
REVIEWERS
This document and summary information on IRIS have received peer review both by
EPA scientists and by independent scientists external to EPA. Subsequent to external review
and incorporation of comments, this assessment has undergone an Agency-wide review process
whereby the IRIS Program Director has achieved a consensus approval among the Office of
Research and Development; Office of Air and Radiation; Office of Prevention, Pesticides, and
Toxic Substances; Office of Solid Waste and Emergency Response; Office of Water; Office of
Policy, Economics, and Innovation; Office of Children’s Health Protection; Office of
Environmental Information; and the Regional Offices.
vi
INTERNAL EPA REVIEWERS
David Reisman
National Risk Management Research Laboratory
Office of Research and Development
Cincinnati, OH
Louis Scarano, Ph.D.
Office of Pollution Prevention and Toxics
Washington, DC
Dharm Singh, D.V.M.
Office of Research and Development
Washington, DC
EXTERNAL PEER REVIEWERS
George Daston, Ph.D.
Miami Valley Laboratories
Procter and Gamble
Cincinnati, OH
Robert Dick, Ph.D.
Consultant
Cincinnati, OH
Sam Kacew, Ph.D.
University of Ottawa
Ottawa, ON
Canada
Summaries of the external peer reviewers’ and public comments, and the disposition of their
recommendations are in Appendix A.
vii
1. INTRODUCTION
This document presents background and justification for the hazard and dose-response
assessment summaries in EPA’s Integrated Risk Information System (IRIS). IRIS Summaries
may include an oral reference dose (RfD), inhalation reference concentration (RfC) and a
carcinogenicity assessment.
The RfD and RfC provide quantitative information for noncancer dose-response
assessments. The RfD is based on the assumption that thresholds exist for certain toxic effects
such as cellular necrosis but may not exist for other toxic effects such as some carcinogenic
responses. It is expressed in units of mg/kg-day. In general, the RfD is an estimate (with
uncertainty spanning perhaps an order of magnitude) of a daily exposure to the human
population (including sensitive subgroups) that is likely to be without an appreciable risk of
deleterious noncancer effects during a lifetime. The inhalation RfC is analogous to the oral RfD,
but provides a continuous inhalation exposure estimate. The inhalation RfC considers toxic
effects for both the respiratory system (portal-of-entry) and for effects peripheral to the
respiratory system (extrarespiratory or systemic effects). It is generally expressed in units of
mg/m3.
The carcinogenicity assessment provides information on the carcinogenic hazard

potential of the substance in question and quantitative estimates of risk from oral exposure and
inhalation exposure. The information includes a weight-of-evidence judgment of the likelihood
that the agent is a human carcinogen and the conditions under which the carcinogenic effects
may be expressed. Quantitative risk estimates are presented in three ways. The slope factor is
the result of application of a low-dose extrapolation procedure and is presented as the risk per
mg/kg-day. The unit risk is the quantitative estimate in terms of either risk per :g/L drinking
water or risk per :g/m3 air breathed. Another form in which risk is presented is a drinking water
or air concentration providing cancer risks of 1 in 10,000; 1 in 100,000; or 1 in 1,000,000.
Development of the hazard identification and dose-response assessment for acetone has
followed the general guidelines for risk assessment as set forth by the National Research Council
(1983). EPA guidelines that were used in the development of this assessment may include the
following: Guidelines for the Health Risk Assessment of Chemical Mixtures (U.S. EPA, 1986b),
Guidelines for Mutagenicity Risk Assessment (U.S. EPA, 1986c), Guidelines for Developmental
Toxicity Risk Assessment (U.S. EPA, 1991a), Guidelines for Neurotoxicity Risk Assessment (U.S.
EPA, 1998a), Draft Revised Guidelines for Carcinogen Risk Assessment (U.S. EPA, 1999),
Guidelines for Reproductive Toxicity Risk Assessment (U.S. EPA, 1996), Recommendations for
and Documentation of Biological Values for Use in Risk Assessment (U.S. EPA, 1988a),
(proposed) Interim Policy for Particle Size and Limit Concentration Issues in Inhalation Toxicity
(U.S. EPA, 1994a), Methods for Derivation of Inhalation Reference Concentrations and
Application of Inhalation Dosimetry (U.S. EPA, 1994b), Use of the Benchmark Dose Approach
in Health Risk Assessment (U.S. EPA, 1995), Science Policy Council Handbook: Peer Review
(U.S. EPA, 1998b, 2000a), Science Policy Council Handbook: Risk Characterization (U.S. EPA,
2000b), Benchmark Dose Technical Guidance Document (U.S. EPA, 2000c) and Supplementary
Guidance for Conducting Health Risk Assessment of Chemical Mixtures (U.S. EPA, 2000d).
The initial literature search strategy employed for this compound was based on the
CASRN and at least one common name. However, because the initial literature search for
“acetone” yielded more than 1,500 hits, subsequent searches were limited to review articles (all
years) and articles with acetone in the title. The following data bases were searched: TOXLINE
(all subfiles), MEDLINE, CANCERLIT, TOXNET [HSDB, IRIS, CCRIS, EMIC (1991-
present), and GENE-TOX], and RTECS. The relevant literature was reviewed through February
2003.
2. CHEMICAL AND PHYSICAL INFORMATION

RELEVANT TO ASSESSMENTS
Chemical and physical properties of acetone are listed in Table 1. Acetone is used
primarily as a chemical intermediate and as a solvent in both chemical and pharmaceutical
applications. Acetone is produced endogenously and utilized in intermediary metabolism
(ATSDR, 1994; WHO, 1998). As the data in Table 1 indicate, acetone is completely miscible
with water and is a highly volatile organic compound.
Table 1. Physicochemical Data for Acetone
Parameter Value Reference
Synonyms 2-propanone; dimethyl ketone Budavari et al., 1996
CAS registry no. 67-64-1
Chemical formula CH3COCH3 Budavari et al., 1996
Molecular weight 58.08 Budavari et al., 1996
Physical state liquid Budavari et al., 1996
Vapor pressure at 20°C 181.72 mm Hg ATSDR, 1994; WHO, 1998
(continued on the following page)
Melting point -94°C Budavari et al., 1996
Boiling point 56.5°C Budavari et al., 1996
Solubility in water miscible Budavari et al., 1996
Log KOW -0.24 ATSDR, 1994; WHO, 1998
Henry’s law constant 4.26 × 10-5 atm-m3/mol ATSDR, 1994; WHO, 1998
Conversion factors in air 1 ppm = 2.374 mg/m3 ATSDR, 1994
Odor threshold in air (absolute) 13-20 ppm ATSDR, 1994; WHO, 1998
3. TOXICOKINETICS
The toxicokinetics of acetone has been studied in part because of the role that acetone
plays in normal metabolism and in disease states (ATSDR, 1994; WHO, 1998). Under starvation
conditions, high-fat and low-carbohydrate diets, or uncontrolled diabetes, fat is metabolized to
form acetoacetate, which in turn is converted to acetone (Wieland, 1968; Argiles, 1986). Under
these conditions, a high level of acetyl CoA generated from the beta-oxidation of fatty acids
coupled with a limited supply of oxaloacetate and a lack of dietary carbohydrate can lead to
ketosis. Much of the research on acetone in humans relates to individuals in these states. Mean
concentrations of acetone in “normal,” nonexposed adult humans have been measured as 840
:g/L in blood, 842 :g/L in urine, and 715 ng/L in alveolar air (Wang et al., 1994). Stewart et al.
(1975) reported blood acetone concentrations in male subjects prior to exposure to acetone as
ranging from 0.73 to 1.29 mg% (mg/100 ml) and in female subjects as 2.86 mg%. Acetone
blood concentrations in healthy individuals are reported to range from 0.3 to 2.0 mg/100 ml
(Physicians Desk Reference, 1976). Owen et al. (1982) reported plasma acetone levels in
diabetics ranging from 1.55 to 8.16 mM (9 to 47 mg%). The normal endogenous acetone
turnover rate (mg/kg-day) is not known.
Overall, available data indicate that humans and rodents readily absorb acetone by
inhalation, ingestion, and dermal exposure. Acetone is broadly distributed throughout the body,
particularly in organs with high water content. The toxicokinetics of acetone are dose-related.
Once acetone has been absorbed, it is extensively metabolized; the prevailing metabolic pathway
appears to be dose-related. At low concentrations, the primary pathway appears to be through
the formation of methylglyoxal. On the other hand, as the concentration of acetone increases,
the propanediol pathway becomes more predominant. Although the second pathway may be
involved in gluconeogenesis, it may also be used to facilitate excretion. Acetone excretion is also
dose-related. In this respect, exposure to low levels of acetone lead to small losses through
expiration. Acetone appears in the urine only when exposure concentrations exceed
approximately 15 ppm. The proportion of acetone lost through expiration increases at high
acetone concentrations.
There is evidence of similarity between the toxicokinetic data on human subjects and
rodents. In both cases, metabolism proceeds by a hepatic pathway at low concentrations and by
an extrahepatic pathway followed by excretion at higher concentrations (Casazza et al., 1984;
Thornalley, 1996). Both species appear to eliminate acetone from the body efficiently (Haggard
et al., 1944; Sakami, 1950; Sakami and Lafaye, 1951; Stewart et al., 1975; Reichard et al., 1979;
Casazza et al., 1984; Wigaeus et al., 1981; Kosugi et al., 1986a; Wang et al., 1994).
3.1. ABSORPTION
The absorption of acetone and its distribution is governed by its physicochemical

parameters and related biological factors. Acetone is miscible in water and has a high vapor
pressure (Table 1) and a high blood/air partition coefficient. The low Kow indicates that acetone
selectively partitions into an aqueous phase rather than a lipid phase; however, acetone is also
slightly lipophilic, allowing for some diffusion into tissues. This suggests that although acetone
is readily absorbed into the aqueous compartments of the body the lipid component may affect
the rate of absorption into the body. Collectively, these factors allow for rapid absorption via the
respiratory and gastrointestinal tracts, and broad distribution throughout the body, particularly
into organs with high water content.
3.1.1. Oral Studies
Acetone is readily absorbed via the oral route, as indicated by early research on humans.
Haggard et al. (1944) administered acetone (50 mg/kg acetone diluted in water; final
concentration and total volume not provided) to male subjects and estimated that between 65 and
93% of the acetone was metabolized while the residual material was excreted from the body over
a period of 2 hours. Both the level of metabolism and excretion through the lungs and urine, and
the short period of time in which these occur indicate that acetone is rapidly absorbed in humans.
Anecdotal information concerning the oral absorption of acetone in humans is provided

in case studies. Studies involving the accidental ingestion of acetone, indicate that acetone is
readily absorbed through the gastrointestinal tract. In one case study (Herman et al., 1997) a 17-
month-old girl was accidently given approximately 4.88 mL/kg of acetone through her
gastronomy tube and was found gagging, nonresponsive, and diaphoretic with dilated sluggish
pupils. Clinical chemistry analyses demonstrated elevated levels of serum ketones. A second
case study (Ramu et al., 1978) involved the accidental ingestion of nail polish remover. The
subject became listless and lethargic with a shortened attention span. While both studies
indirectly demonstrate that acetone is readily absorbed via the gastrointestinal tract a quantitative
assessment of absorption cannot be determined.
Rapid absorption of acetone in rodents was demonstrated by Price and Rittenberg (1950).
14
When rats were administered 0.22 mg of C-acetone (1.2 mg/kg) in water, 47% of the acetone
14
was expelled as CO2 within 13.5 hours following administration (29% released within the first
2.5 hours). In a second experiment in which rats were administered pulses of 14C-acetone on a
daily basis over a 7.5-day period, 67 to 76% of the administered acetone was expelled as 14CO2
over the ensuing 24-hour period. The measurements started with short intervals extending to 6
hours during the first day of the study, with 56% of the administered radiolabel appearing during
the initial 6 hours. On subsequent days, measurements were made at 6-hour intervals. The
testing scheme does not support comparison with the initial study, which demonstrated high
levels of absorption and rapid metabolism with successive applications of the chemical on the
rate of absorption. The expiration of acetone was determined following the administration of
14
C-acetone (only 7% of the administered acetone was expelled in the breath, with peak levels
achieved 2 hours following administration). Overall, the data indicate that acetone is rapidly
absorbed into the body upon ingestion. The acetone that is expelled in the breath is in the
unmetabolized form or CO2.
3.1.2. Inhalation Studies
The most important physicochemical parameters relating to exposure to acetone are the
vapor pressure and Henry’s constant. Acetone has a high blood-air partition coefficient (KB/A) of
301 (Dills et al., 1994; Wigaeus et al., 1981; ATSDR, 1994; WHO, 1998). However, a range of
partition coefficients has been reported, including 245 by Sato and Nakijima (1979) and 210 by
Hallier et al. (1981). The high KB/A indicates that acetone is rapidly absorbed into the body via
the inhalation route. During inspiration, acetone passes through the epithelial cells in the nasal
cavity and is dissolved into the bloodstream where it is transported. The fraction of acetone
remaining in the nasal tissue evaporates during expiration into the environment. The differences
between the concentration of acetone during inhalation and exhalation accounts for the portion
that is absorbed and dissolved into the bloodstream (Dahl et al., 1991).
Hallier et al. (1981) conducted studies using rats in desiccators to achieve maximum
saturation for determining the partition coefficients. Rats were placed in a desiccator that was
saturated with acetone. Measurements of acetone in the head space were made over time and the
pharmacokinetic parameters were determined by loss of acetone. The equilibrium constants
(Keq) were determined to be 330, 210, and 220 for urine/air, blood/air, and whole animal/air
ratios, respectively. The Keq for oil/air ratio was 125. By measuring the amount of acetone taken
up by the rats in the desiccator, it was determined that the maximum amount of acetone absorbed
slightly exceeded the water pool of the rodents. The authors proposed that the limited
lipophilicity of acetone contributes to a minor amount of nonmetabolized acetone that moves
into the lipid fraction of tissues.
Biological conditions such as the rate of respiration and the blood/air partition coefficient
have also been shown to be key factors in the uptake of acetone via the inhalation route. Wigaeus
et al. (1981) exposed male volunteers to 1,300 mg/m3 (547.6 ppm) acetone at rest or 700 mg/m3
(294.9 ppm) with exercise for 2 hours. The total amount taken up increased as ventilation
increased with exercise, but the relative uptake ranged from 39-52% for both exposures. Total
uptake was 0.6 and 1.2 g, respectively, for the 1,300 mg/m3 (resting) and 700 mg/m3 (active)
exposures. Acetone concentrations in alveolar air were 30-40% of those in inspiratory air. This
fraction did not change with exposure time or workload. In contrast, acetone concentrations in
blood increased continuously during exposure, with no sign of equilibrium between
concentrations of inspired air and blood. The constant rate of absorption by inhalation, in
contrast to the continuous increase of acetone in the blood, most likely reflects the high KB/A in
conjunction with its movement from the nasal cavity and distribution throughout the body. Given
the high KB/A of acetone, it is surprising that only about 40% of the administered acetone is
absorbed into the body. This may be due to the relatively low lipid solubility of acetone, which
provides resistance to the transfer of acetone from the air through the nasal tissue and into the
bloodstream. At the end of the 1,300 and 700 mg/m3 exposures, acetone concentrations in
arterial blood were 15 and 75 mg/kg, respectively. Acetone concentrations in alveolar air
dropped rapidly during the first 5 minutes following exposure. Half-times for acetone in
alveolar air, arterial blood, and venous blood averaged 4.3, 3.9, and 6.1 hours, respectively.
Elimination of acetone via the lungs was about 20% of total uptake, whereas only about 1% of
uptake was excreted via the urine.
Other studies support the findings of Wigaeus et al. (1981) for the uptake and elimination
of acetone in humans. Mean relative acetone uptake averaged 53% in volunteers exposed to 21-
211 ppm acetone for up to 4 hours at rest or 2 hours with intermittent exercise (Pezzagno et al.,
1986). Acetone concentrations in the capillary blood of male volunteers were shown to increase
steadily during a 2-hour exposure to 231 ppm acetone followed by a monoexponential
postexposure decay curve with a calculated half-life of 4.3 hours (Ernstgård et al., 1999).
Although there are only a few studies specifically measuring the uptake of acetone by
inhalation, a number of studies demonstrate indirectly that acetone is rapidly absorbed via
inhalation. For example, Wigaeus et al. (1982) measured the distribution of acetone in mice
following inhalation exposure to 1,200 mg/m3 (500 ppm) 2-14C-acetone. The study did not
involve a quantitative measure of the total amount of distributed acetone, but did measure the
distribution of acetone in individual organs. The study demonstrated a continuous increase in
the tissue concentration of acetone and total radioactivity during the first 6 hours of exposure.
This study is discussed in greater detail in Section 3.2.
Similar results have been found in dogs and rats. Acetone uptake by the respiratory tract
of the dog was 42-54% for exposure concentrations of 0.36-0.80 :g/mL (150-340 ppm) for an
unspecified duration (Egle, 1973). Blood concentrations of acetone in rats exposed to 150 ppm
increased gradually over a 2-hour period, then reached a plateau at steady state for the remainder
of the 4-hour exposure (Geller et al., 1979a). Plasma acetone levels in female rats corresponded
with increasing exposure concentrations, but were not related to pregnancy status or gestation
time (Mast et al., 1988).
3.1.3. Dermal Studies
Dermal absorption of acetone has been shown to occur rapidly in humans. In a Japanese
study (Fukabori et al., 1990) translated and described by ATSDR (1994) and WHO (1998),
cotton soaked with acetone was applied to the skin of volunteers for 2 hours/day for 4 days (dose
information not provided). Resulting levels of acetone were 5-12 mg/L in blood, 5-12 ppm in
alveolar air, and 8-14 mg/L in urine. When the daily exposure was increased to 4 hours, the
body burden more than doubled. Absorption was immediate, with peak levels occurring at the
end of each application. Concomitant inhalation exposure is expected from dermal exposure.
From the alveolar air and urine concentrations, the study authors calculated that 2- and 4-hour
dermal exposures were equivalent to 2-hour inhalation exposures of 50-150 ppm and 250-500
ppm, respectively.
Studies concerning the level of dermal absorption of acetone in rodents are limited,
although acetone is frequently used as a vector in dermal studies of other chemicals (NTP, 1991,
1995, 1997). However, no available studies included an acetone control and a naive control.
Together, these controls could be used to directly measure acetone uptake into the body via the
dermal route.
3.2. DISTRIBUTION
Tissue distribution of 2-14C-acetone following inhalation exposure to 1,200 mg/m3 (500

ppm) was studied in mice (Wigaeus et al., 1982). Acetone in blood, lung, kidney, brain,
pancreas, spleen, thymus, heart, testis, vas deferens, muscle, and subcutaneous and
intraperitoneal white adipose tissue reached steady state concentrations within 6 hours and
showed little or no accumulation when exposures were prolonged up to 24 hours or repeated 6
hours/day for 5 days. In contrast, radioactivity in liver and brown adipose tissue continued to
increase during a 24-hour exposure and increased in adipose tissue with repeated exposures. Of
all studied tissues, the liver contained the highest level of radioactivity and the adipose tissues
the lowest. Only about 10% of the radioactivity in the liver was unchanged acetone. Half-times
for acetone elimination were 2-3 hours for blood, kidney, lung, brain, and muscle, and greater
than 5 hours for subcutaneous adipose tissue. By 24 hours after exposure, acetone
concentrations had returned to endogenous levels in all tissues. The study supports older data
(Haggard et al., 1944) that acetone distributes evenly in body water and does not accumulate
with repeated exposure. The accumulation of radioactivity in liver and brown adipose tissue
may reflect high metabolic turnover in these tissues.
Scholl and Iba (1997) measured the distribution of acetone in rats following acetone
inhalation. Male Sprague-Dawley rats were exposed to 1,000 ppm (2,400 mg/m3) of acetone for
3 hr/day for 10 days. Tissue concentrations of acetone were determined 1 h following the final
exposure. Mean concentrations of acetone in the plasma, liver, lung, and kidney were 35.3, 13.2,
11.4, and 21.8 :g/g, respectively. The concentration of acetone in plasma relative to the three
organs approximates the octanol-water partition coefficient (log Ko/w = -0.24) and reflects the
greater solubility in water, which is the major constituent of plasma, compared with lipid
solubility.
3.3. METABOLISM
Acetone has been broadly studied as a metabolic intermediate that is naturally formed in
humans and rodents under normal metabolic conditions and at higher concentrations under
conditions of fasting, ingestion of high-fat, low-carbohydrate diets, and uncontrolled diabetes.
The proposed metabolic pathways for acetone are shown in Figure 1. Reichard et al. (1979)
studied the metabolism of acetone in 15 human subjects undergoing starvation ketosis to
evaluate the role of acetone in starvation-induced ketonemia in humans. Acetone radiolabeled
(14C) on the carbonyl carbon was administered to measure the amount of acetone expired in the
breath, excreted in the urine, and metabolized to CO2. The study used human subjects of both
sexes ranging from 22 to 52 years of age. The subjects were divided by weight into obese and
nonobese categories where the nonobese individuals were -7 to +16% and obese individuals
were +38 to +155% of the ideal body weight (based on Metropolitan Life Insurance tables) at the
start of the study. The obese individuals were starved for either 3 (n=6) or 21 (n=3) days and the
nonobese were starved for 3 days (n=3). Following administration, periodic measurements were
taken of the subjects’ expired air and the amount of 14CO2 was determined. On the day 14C-
acetone was administered, hourly measurements were taken over a six-hour period for total
glucose, acetoacetate, $-hydroxybutyrate, acetone in plasma and urine. The amount of radiolabel
in each constituent was used to determine the metabolism of acetone.
Of the studied components, the radiolabel was found in 14CO2, acetone and glucose.
While the overall concentration of plasma acetone remained constant over the six-hour period,
the amount (specific activity) of 14C acetone decreased and the amount (specific activity) of 14C
glucose and 14CO2 increased indicating that acetone is metabolized into glucose which is
subsequently metabolized to CO2. No radiolabel was found in $-hydroxybutyrate, acetoacetate,
or plasma free fatty acids indicating that while ketoacids are metabolized to acetone, there is no
reverse reaction that accounts for an increase in ketoacids and an exacerbated ketotic state. The
absence of radiolabel in fatty acids indicates that starvation conditions are not conducive to
lipogenesis. The study also found that the ratio of acetone in expired air and urine is proportional
to the plasma concentration in humans. The principal findings in these studies were that,
depending upon the plasma acetone concentration, excretion of acetone from the body in breath
and urine accounts for about 2 to 30% of the endogenous acetone under fasting conditions.
Conversion of acetone to other biological compounds is the primary mode of acetone
elimination. The authors estimate that 50-70% of the acetone that was eliminated in 3-day fasted
subjects was lost through metabolism. On the basis of the specific activity data, approximately 4-
11% of plasma glucose production could theoretically be derived from acetone.
Owen et al. (1982) evaluated the metabolism of ketone bodies and the toxicokinetics of
acetone in diabetics. The study consisted of selecting nine patients that the study authors
characterized as “acutely ill” with acidemia, hyperglycemia, and hyperketonemia. The patients
were dosed with 0.75-1.56 :mol of 2-14C-acetone intravenously. Blood samples were taken
every 1-2 hours for up to 10 hours. Breath and blood samples were taken simultaneously.
During the study, insulin therapy was dictated by the patients’ clinical situation. The mean time
between acetone injection and insulin administration was 3 hours with a range of 1-6 hours.
Analyses were conducted for acetone, $-hydroxybutyrate and acetoacetate in plasma, breath and
urine, and for glucose in plasma.
A mean plasma acetone concentration of 4.96 mM with a range of 1.55-8.91 mM was

determined at the start of the study. The higher acetone concentrations in this cohort are
consistent with patients exhibiting severe ketoacidosis while the lower levels are similar to
values seen in healthy, fasting individuals. The data demonstrates a relationship between the
plasma acetone levels and levels of acetone in the breath (r=0.95, p<0.001) and, to a lesser extent
urine (r=0.68, p<0.05). In one subject 14C-acetone declined at a linear rate while the specific
activity of 14CO2 increased, reaching peak levels at 6 hours following dosing and declining
thereafter. However, overall 14C radiolabel was detected in the protein of all patients and in the
lipids of six of the nine subjects, albeit at low levels. The radiolabel was found in glucose in
only 3/9 subjects; these were the individuals with the lowest levels of plasma acetone at the start
of the study. Although the authors determined an average acetone turnover rate of 265±47
:mol/min/1.73 m2, there was no relationship between the individual acetone turnover rate and
plasma acetone concentration as has been observed in fasting individuals.
Plasma concentrations of acetoacetate and $-hydroxybutyrate fluctuated considerably

while the concentration of acetone remained constant reflecting a steady state condition. This is
apparently achieved through a combination of metabolism and excretion. A positive linear
relationship between the blood concentration of acetone and the amount lost in the breath was
noted, and a negative relationship with in vivo metabolism. For example, at “low” (1-2.5 mM)
plasma acetone concentrations ~20% of the acetone was lost through expired air and ~75% was
lost through metabolism, while at “high” (8-9 mM) concentrations ~80% was lost through
expired air and ~20% was lost through metabolism. Urinary excretion of acetone accounted for
about 7% of the acetone excreted, independent of plasma acetone concentration. These data
show that the percentage of acetone exhaled in breath increases and the percentage metabolized
diminishes as acetone plasma concentration increases. The 14C radiolabel on the carbonyl carbon
10
of acetone was detected in glucose, protein and lipids while no label was detected in $
hydroxybutyrate, acetoacetate or free fatty acids.
The same authors conducted a follow-up study (Reichard et al., 1986) to elucidate the
pathway involved in the metabolism of acetone to glucose. Seven patients with diabetic
ketoacidosis received an infusion of 2-14C-acetone over a 6-hour period. Patients were not
maintained on insulin therapy during the study. The authors monitored the incorporation of the
radiolabel into acetol and 1,2-propanediol, and subsequently into glucose. The data
demonstrated a linear relationship between acetol and propanediol formation and plasma
acetone. In subjects with lower plasma acetone concentrations the acetol concentration was
higher than propanediol, while at higher acetone concentrations, the acetol level was lower than
propanediol. The study demonstrates the incorporation of the label into glucose. Interestingly,
in six of the seven patients the label was incorporated predominantly into the 1, 2, 5, and 6
carbons of glucose while in one patient the label was found predominantly in the 3 and 4
carbons. The data suggest two separate metabolic pathways for converting acetone to glucose.
The patient in which the resulting glucose was predominantly labeled in the 3 and 4 positions
also had the highest level of endogenously produced acetone at the start of the study. The
relative amount of intermediates (acetol and propanediol) may suggest that the predominant
pathway is via the methylglyoxal pathway at low acetone concentrations, while the propanediol
pathway predominates at higher levels. The study does not report changes in glucose levels with
the infusion of acetone.
The average plasma acetone turnover rate was 533 µmol/min/1.73m2, a value twice that
obtained in the Owen et al. (1982) study via the single injection technique. In addition, plasma
acetone turnover rates were directly related to the plasma acetone concentrations, unlike the
results of the Owen et al. (1982) study. The authors state that the single injection technique may
have resulted in inadequate mixing of the labeled acetone in the body pools of acetone and may
have been responsible for the failure to show a linear relationship between plasma acetone
concentrations and acetone turnover rates.
Sakami (1950) conducted a series of rat studies to determine the metabolism of

exogenously administered acetone. The first study involved starving the rats for 2 days followed
by gavaging with 14C-labeled acetone and measuring the generation of 14CO2. The rats were
dosed and placed in metabolic chambers where the gas was sampled on an hourly basis. The
animals received additional dosing of 14C acetone at 3-, 6-, and 10-hour samplings.
Radioactivity trapped by the bicarbonate was 14CO2 indicating acetone is metabolized to CO2.
11
While the study does not describe the method for trapping CO2, it is possible that the
radioactivity that was analyzed was nonmetabolized expired acetone.
In the same study, rats were sacrificed following incubation and necropsied, and the liver
homogenized. Serine, glycogen, methionine, and choline fractions were recovered and assayed
for radioactivity. Of the fractions tested, 14C was predominantly found in the glycogen that was
recovered from the liver, indicating a metabolic process that involves the conversion of acetone
to glucose. Extracts from the viscera of the rats contained radioactivity in serine, methionine, and
choline indicating that formate contributes to their synthesis at the same positions within the
molecule as those found in the labeled constituents. Hence, the authors propose that acetone is
converted to serine, methionine, and choline through the oxidation and decarboxylation of
acetone to acetate and formate. The authors do not speculate on a pathway. Although they may
be correct in stating that the 14C label is incorporated into the amino acids, the authors do not
provide support for the contention that formate is an intermediate in the process. It should be
noted that methionine is an essential amino acid in humans.
As a follow-up to their initial work, Sakami and Lafaye (1951) conducted a study to
elucidate the pathway for acetone metabolism. The authors proposed a method for elucidating
the metabolic pathway based on the locations of 14C in glucose that is synthesized from acetone.
If the metabolism of 2-14C-acetone proceeded through the cleavage of acetone to acetate and
formate, the acetate would form acetyl CoA, which would lead to higher labeling at the 3- and 4-
positions relative to the 2- and 5-positions. Six male rats were fasted for 24 hours and gavaged
with glucose and 2-14C-acetone and placed in a metabolic cage for 4 hours. The amount of 14CO2
generated from acetone increased with time over the course of the incubation period. At the end
of 4 hours the animals were sacrificed, livers were removed, and glycogen was extracted. The
glycogen was digested to glucose and subjected to microbial degradation to identify sites of 14C
labeling on the glucose molecule. The 14C label was found in all positions in the glucose
molecule, although with higher labeling in the 2- and 5-positions than the 1- and 6-positions.
The authors indicate that the level of labeling in the 1- and 6- labeled molecules may be an
artifact of the degradation process.
Although the data from Sakami (1950) and Sakami and Lafaye (1951) provide strong
evidence that exogenous acetone is readily absorbed via the gastrointestinal tract and converted
to CO2, glucose/glycogen, and other metabolites, the studies fail to provide adequate support to
define a metabolic pathway.
12
On the basis of findings that acetone is converted to glucose in the body, Casazza et al.
(1984) set out to describe the metabolic pathway(s) for the conversion of acetone to glucose or
other cellular constituents. They conducted studies that employed a combination of whole
animals, hepatocyte cultures, and microsomes from rat livers to develop a composite pathway for
the metabolism of acetone in the rodent. Pretreatments included rats that received either
nontreated drinking water or drinking water with 1% acetone up to 14 hours prior to sampling,
when the acetone-treated water was replaced with nontreated water. Rats from both groups
received either an intraperitoneal treatment of saline or saline with 5 :mol/g of acetone. Rats
that had been pretreated with 1% acetone in drinking water up to 14 hours prior to sampling and
had received an intraperitoneal injection of acetone produced detectable levels of 1,2-
propanediol and 2,3-butanediol. Rats that received no intraperitoneal injection or received the
intraperitoneal injection with no pretreatment with acetone failed to produce detectable levels of
either of the two diols, indicating the role of enzyme induction in determining a particular
pathway in the metabolism of acetone.
In the same study, elevated levels of lactate were found in rats that received pretreatment
of acetone compared with rats receiving drinking water in the nontreated and treated groups. To
determine the metabolic pathway leading to lactate, acetone was incubated in vitro in a
microsomal suspension and acetol was recovered. When acetol was added to the microsomal
suspension in the presence of glyoxalase I and glutathione, D-lactate was recovered, but in the
absence of glyoxalase I and glutathione, methylglyoxal was recovered. Incubation of rat
hepatocytes with acetol or methylglyoxal produced D-lactate and glucose. However, when rats
were pulsed with 14C-acetone and supplemented with D-lactate the incorporation of 14C into
glucose was not reduced, indicating that the predominant route of gluconeogenesis is not via the
formation of D-lactate, but directly from methylglyoxal through glyoxalase(s). By analyzing the
kinetics of the conversion of acetone or acetol to glucose and D-lactate, the rate of utilization of
acetol could not be accounted for by the formation of glucose and D-lactate. These findings
suggest the presence of an alternate pathway for the conversion of acetone to glucose.
Further studies (Casazza et al., 1984) were performed using hepatocytes from a rat
starved for two days given 1% acetone in drinking water. The hepatocytes were incubated with
L-lactate and L-1,2-propanediol with and without specific inhibitors of aldehyde dehydrogenase
(cyanamide) or alcohol dehydrogenase (pentylpyrazole). The rate of glucose formation from L-
lactate was reduced 12 and 50% in the presence of pentylpyrazole or cyanamide, respectively,
compared with the control. The rate of glucose formation from L-1,2-propanediol in the
13
presence of pentylpyrazole or cyanamide was comparable to the rate of glucose formation in the
controls (without L-1,2-propanediol).
A comparison of the kinetics for L-1,2-propanediol production from hepatocytes with

those observed in the whole rat indicates that the enzymatic activity in the liver could not
account for the total amount of L-1,2-propanediol formed in the whole animal. Additionally,
1,2-propanediol concentrations were comparable in the hepatic artery and vein of perfused liver
containing acetol. The data suggest that most of the metabolism occurring via the formation of
1,2-propanediol is extrahepatic and that 1,2-propanediol production provides a second pathway
for the formation of glucose from acetone.
In summary, Casazza et al. (1984) provide evidence that there are two pathways (hepatic
and extrahepatic) involved in the metabolism of acetone to glucose. The hepatic route involves
the conversion of acetone to acetol mediated by acetone monooxygenase, which has been shown
to be CYP2E1. Acetol in turn may be converted to methylglyoxal via the CYP2E1 enzyme
system or converted to 1,2-propanediol. Methylglyoxal is converted either directly to glucose or
to D-lactate, which is converted to glucose. The data support the direct pathway of
methylglyoxal to glucose as opposed to the D-lactate route. From the data presented in this
study, it is not possible to determine which of the two pathways, i.e., the 1,2-propanediol or
methylglyoxal pathway, predominates.
Although the studies are comprehensive, they have notable limitations. Treatments for
most of the assays were not replicated, and, therefore, provide no measure of variability. In
addition, the authors note that formate and acetate production has been proposed as a pathway
for the metabolism of acetone; however, this issue is not addressed in their paper. It is not clear
whether the authors analyzed for an “active” form of acetate or formate and failed to find it.
Kosugi et al. (1986a) demonstrated that the predominant pathway for the metabolism of
acetone is dose-dependent, with the methylglyoxal pathway predominating at lower
concentrations and the 1,2-propanediol pathway predominating at higher concentrations. These
studies support the human studies in diabetics (Reichard et al., 1986). Rats were fasted for 20-24
hours (except for two controls) and then received either “trace” amounts or 1.6 mmol of 2-14C-
acetone through a tail-vein catheter. 13C-lactate was administered via the same route. “Trace”
amount was defined as the specific activity of the undiluted 2-14C acetone as it was received
from the distributor. The infusion of acetone took approximately four hours to complete.
Following the infusion of radiolabeled acetone, a sample of blood was drawn, the animals were
14
sacrificed, and the disposition of the 14C- label was determined. Rats were either necropsied and
the livers removed to recover the glycogen that was digested to glucose, or they were frozen and
powdered and the glucose recovered. For all samples, the glucose was degraded and analyzed
for the placement of 14C in the glucose molecule. Regardless of whether the rats were fasted or
infused with glucose and acetone, the infusion of trace amounts of 2-14C acetone into the rat
resulted in 5-10% of the radiolabel in the 3 or 4 positions of the glucose molecule. In rats that
received the 1.6 mmol acetone solution, 23-40% of the 14C label was in either the 3 or 4 positions
on the resulting glucose. Differences in the location of 14C indicate that there is more than one
metabolic pathway and that the pathway selection is dose-dependent.
Kosugi et al. (1986b) conducted a second study comparing the metabolism of

radiolabeled 2-14C-lactate with that of 2-14C-acetone. The study authors proposed that if acetone
is metabolized prior to any randomization of the carbons in which lactate serves as an
intermediate, as postulated by Casazza et al. (1984), the resultant glucose molecule should be the
same as that seen with lactate, with most of the 14C in carbons 1, 2, 5, and 6 of the glucose
molecule. Therefore, differences in 14C distribution in the glucose molecule between the two
levels of acetone administration indicate that in low concentrations acetone is metabolized
through a pathway that involves lactate as an intermediate, but at high levels acetone is
metabolized through a pathway that does not involve lactate as an intermediate or byproduct.
The authors postulate that the catabolism of glucose most likely proceeds through the formation
of acetate as proposed by Sakami (1950).
Gavino et al. (1987) conducted studies to clarify the proposed pathways by which
acetone is utilized at millimolar concentrations. Sprague-Dawley rats were starved for 48 hours
prior to the start of the study. Livers were perfused with glucose and bovine serum albumin for
30 minutes, after which 2-14C-acetone was added to the perfusate. The data demonstrate that as
the acetone concentration decreased, the concentration of acetoacetate increased. The perfusate
was analyzed for the incorporation of 14C into potential metabolites including citrate, lactate, $
hydroxybutyrate, acetate, 1,2-propanediol, and acetol. The 14C radiolabel was found in glucose,
CO2 and volatiles, but not in $-hydroxybutyrate indicating that acetone is not carboxylated.
Although the authors acknowledge that considering the proximity of elution there might be some
incorporation of 14C into 1,2-propanediol, most of the 14C that was introduced into the perfusate
as 2-14C-acetone was found in acetate with the radiolabel in the 1 position.
Casazza et al. (1984) proposed two main pathways of acetone metabolism: one
proceeding via methylglyoxal and a second through 1,2-propanediol. Kosugi et al. (1986a)
15
provide evidence that the route of metabolism differs depending on the amount of acetone. For
example, at lower concentrations metabolism proceeds mostly through the methylglyoxal
pathway, but as the concentration of acetone increases catabolism proceeds to a correspondingly
greater degree via the 1,2-propanediol pathway. Results of the Gavino et al. (1987) study
indicate that the catabolism of acetone proceeds via a pathway that generates acetate but not via
1,2-propanediol. However, a significant difference between the two studies is that Kosugi et al.
(1986a) administered acetone via a tail cannula, which subjected the acetone to systemic
metabolism. Gavino et al. (1987), on the other hand, conducted a hepatic perfusion study. The
differing results are consistent with the mechanism proposed by Casazza et al. (1984), who
stated that the methylglyoxal pathway was largely a hepatic pathway and the 1,2-propanediol
pathway was largely extrahepatic.
Therefore, the metabolic pathway(s) for acetone is proposed to have three possible
routes, all of which start with the conversion of acetone to acetol. From acetol the pathway
diverges to the formation of either methylglyoxal or 1,2-propanediol. A third pathway that has
been proposed involves the production of formic acid which is the metabolite that is mostly
responsible for the toxic effects resulting from consumption of methanol. Following ingestion,
methanol is readily converted to formaldehyde and subsequently to formic acid. Under
conditions of methanol poisoning, formic acid inhibits the formation of cytochrome c oxidase,
which causes histotoxic hypoxia leading to several adverse effects. The most sensitive effect is
amblyopia and amaurosis, a condition of the optic nerve causing impaired vision or blindness.
At higher levels, formic acid affects other organs, particularly those with high oxygen
consumption rates such as the brain, heart, and kidneys. At high enough levels, methanol
poisoning will cause death, presumably due to the formation of formic acid (Liesivuori and
Savolainen, 1991).
Data supporting the formation of formic acid as a product of acetone metabolism are
sparse. Hallier et al. (1981) found that rats placed in a desiccator saturated with acetone
demonstrated low (4.7% of the absorbed acetone) amounts of formate in the urine. In a recent
review of the literature concerning the metabolism of acetone, Kalapos (1999) indicates that
enzyme systems have not been identified to mediate the formation of formate from acetone, and
that the toxic effects attributed to acetone are inconsistent with those of formic acid.
Methylglyoxal is an "-oxoaldehyde formed from the metabolism of acetone, and has

been shown in rodents to be catabolized to lactic acid through glyoxalase I and II and
glutathione, or metabolized directly into a gluconeogenesis route by an undetermined pathway.
16
Methylglyoxal has been shown to have potentially adverse effects at levels higher than normally
seen in the body, including: genotoxicity (Barnett and Munoz, 1998), depletion of glutathione
(Ankrah and Appiah-Opong, 1999), and the induction of apoptosis (Thornalley, 1998).
Methylglyoxal has also been shown to induce sex-linked recessive lethal responses in
Drosophila melanogaster germinal cells, although only at high concentrations. Depletion of
glutathione resulting from the catabolism of methylglyoxal to lactate may be significant. Ankrah
and Appiah-Opong (1999) demonstrated that perinatal exposure to methylglyoxal altered the
tolerance of mice to glucose, while postnatal blood analyses demonstrated a decreased amount of
glutathione-S-transferase and a decreased ability of red blood cells to tolerate oxidative stress.
Finally, methylglyoxal has been shown to arrest growth and induce apoptosis in human leukemia
cells (Kang et al., 1996).
17
H3C CH3
Acetone
acetone
monooxygenase
CYP2E1
H no system OH
O acetol kinase, L1,2 propanediol C OH HC
H3C C
OH dehydrogenase, glycerol P phosphatase OHH identified
H3C C 2 O
H2 L-1,2-Propanediol Formic acid
Acetol alcohol
lactaldehyde dehydrogenase,
acetol reductase O
monooxygenase
CYP 2E1
H H3C OH
O C O
methylglyoxal reductase C Acetic acid
H H3C
OH H
H3C
Lactaldehyde
O
Methylglyoxal lactaldehyde
dehydrogenase
methyl acyl CoA
glyoxalase I synthase
GSH alpha-oxoaldehyde (thiokinase)
dehydrogenase H OH
C
H H3C
C OH
SG O
H3C
OH L-lactic acid
S-D-Lactoylglutathione L-lactate
dehydrogenase
methyl O
glyoxalase II OH O
H3C
O
OH Pyruvic acid H3C CoA
pyruvate dehydrogenase acetyl-CoA
C OH complex
H3C H D-2-hydroxyacid
dehydrogenase
O
D lactic acid
Citrate
O O
HO
C OH TCA cycle
gluconeogenesis H2
Glucose O
Oxaloacetic acid
(after Kalapos, 1999)
18
Figure 1. Pathway for the metabolism of acetone.

Bondoc et al. (1999) demonstrated the role of CYP2E1 in the metabolism of
endogenously-produced acetone under ketogenic conditions using a knockout mouse model
lacking the ability to express CYP2E1. When mice were fed ad libitum, plasma acetone levels in
strain-matched CYP2E1-competent mice, and CYP2E1-null mice had comparable levels of
plasma acetone. After fasting for 48 hours, the competent strain had a 2.5- to 4.4-fold increase in
plasma acetone while the CYP2E1-null mice had a 48-fold increase. This study provides strong
evidence for the role of CYP2E1 in the catabolism of acetone.
In brief, based on human and animal studies and in vitro studies, the metabolism of
acetone may occur via at least two routes (Figure 1). The principal metabolic pathways are
dependent on the site of metabolism and on the concentration of acetone. The metabolites are
incorporated into glucose and other substrates of intermediary metabolism that ultimately
produce CO2. In the first metabolic step, common to all potential pathways, acetone is oxidized
to acetol by acetone monooxygenase, an activity associated with CYP2E1. This step requires O2
and NADPH (Casazza et al., 1984). In the first pathway, acetol is converted to methylglyoxal,
which in turn is metabolized to glucose through a lactate intermediate. The conversion of
acetone via the methylglyoxal pathway is mediated by acetone monooxygenase (CYP2E1) and
acetol monooxygenase (CYP2E1) to form methylglyoxal. The conversion of methylglyoxal to
lactate is mediated by glyoxylase I and II and glutathione-S-transferase. This pathway is
primarily a hepatic pathway. In the second pathway, the acetol intermediate is converted to L-
1,2-propanediol by an extrahepatic mechanism that has not been fully characterized. The
metabolism of acetone via the 1,2-propanediol pathway to lactate is mediated by alcohol
dehydrogenase and aldehyde dehydrogenase (Dietz et al., 1991). Gluconeogenesis may proceed
through the formation of an active form of acetate. 1,2-Propanediol may be converted to glucose
through a series of intermediates including lactate. As noted above, Sakami (1950) proposed a
pathway by which acetone is converted to formate and acetate.
The data also demonstrates that the pathways for acetone metabolism are concentration-
dependant. At lower concentrations, acetone is metabolized in the liver through the
methylglyoxal pathway similar to biological conditions of fasting or exertion where the acetone
is formed from fatty acids to produce glucose. Thus, at low plasma concentrations acetone serves
as a gluconeogenic substrate. At higher concentrations an alternate pathway predominates and
mediates the conversion of acetone to 1,2-propanediol. Although some studies indicate that 1,2-
propanediol serves as an intermediate in the production of glucose, it is conceivable that the
conversion from acetone to the diol diverts acetone from gluconeogenesis and facilitates the loss
of acetone via urine.
19
Enzymes involved in the metabolism of acetone are inducible. The metabolism of

acetone through the methylglyoxal route is mediated largely by CYP2E1, which can be induced
by fasting, experimental diabetes, or exposure to ethanol or acetone; therefore, acetone induces
its own metabolism (ATSDR, 1994; Mandl et al., 1995; WHO, 1998). Inhibition of CYP2E1
activity resulted in an increase in endogenous acetone levels in rats (Chen et al., 1994). Acetone
significantly increased both the microsomal protein content and the activity of CYP2E1 in rat
liver 18 hours after a single oral dose of 15 mmol/kg body weight (Brady et al., 1989) and in
mouse liver 24 hours after a single oral dose or administration of 1% in the drinking water for
eight days (Forkert et al., 1994). Treatment with acetone or starvation conditions leads to
increases in protein content and enzyme activity in the rat kidney (Ronis et al., 1998). Acetone
inhalation exposure has also been shown to potentiate enzyme induction by the solvents toluene
and xylene (Nedelcheva, 1996).
3.4. EXCRETION
In a previously described study, Wigaeus et al. (1981) exposed male subjects to two
concentrations of acetone, with and without exercise. Individuals at rest were exposed to 1,300
mg/m3 and on a second occasion to 700 mg/m3 either with light exercise or with increasingly
strenuous exercise for two hours. As noted earlier, inhalation uptake increased with exertion.
The exposure treatment was followed by a 4-hour monitored period of elimination consisting of
intermittent walking and rest during which expiratory samples and urine were collected and
analyzed for nonmetabolized acetone. The highest concentration of acetone in urine was found
between 3 and 3.5 hours following exposure. Absorbed acetone was lost through the lungs in the
form of nonmetabolized acetone (16, 20 and 27% for resting, light exercise, and more strenuous
exercise, respectively). For all three exposure scenarios, approximately 1% of the absorbed
acetone was lost through the kidneys, the remainder was presumed to be metabolized. The
higher concentration of acetone that was lost through the lungs corresponds to a greater amount
of acetone absorbed by the body, suggesting saturation of acetone metabolism.
In male volunteers given oral acetone doses of 40-80 mg/kg, an estimated 65-93% of the
dose was metabolized, with the remainder being eliminated in the urine and expired air in about
two hours, indicating rapid and extensive absorption by the gastrointestinal tract (Haggard et al.,
1944).
Wang et al. (1994) found that workers with a mean occupational exposure to acetone of
141.8 ppm had blood and urine concentrations at the end of a shift of 23 mg/L and 22 mg/L,
20
respectively; acetone concentrations remained slightly elevated 16 hours after the end of the shift
and the blood half-life was calculated to be 5.8 hours. A positive linear correlation has been
shown between acetone concentrations in the breathing zone of workers and urinary (Kawai et
al., 1992), blood (Wang et al., 1994), and alveolar concentrations (Wang et al., 1994). Breath
decay curves from experimentally-exposed individuals have been shown to be highly
reproducible, and the narrow range of acetone in the breath at a specific postexposure time
indicates that breath analysis could be a reliable method to estimate the magnitude of recent
acetone exposure by inhalation (Stewart et al., 1975). Urine concentrations, on the other hand,
were shown to increase only when workers were exposed to acetone concentrations greater than
15 ppm (Kawai et al., 1992). Differences between the findings of Wang et al. (1994) and
Wigaeus et al. (1981) may reflect dose-related differences, given that the subjects in the Wigaeus
et al. (1981) study were exposed to acetone concentrations that were roughly five-fold higher
than those in the Wang et al. (1994) study.
Haggard et al. (1944) injected 11 rats intraperitoneal and followed the fate of acetone
over a 4- to 6-hour period. Blood acetone levels and the amount of acetone lost through the
lungs and urine were compared with that lost by metabolism. The data indicate that the percent
of acetone lost by elimination in the urine and expiration is directly proportional and that lost via
metabolism is inversely proportional to the blood acetone content. At higher blood
concentrations acetone is predominantly lost via elimination, whereas at low concentrations it is
metabolized. Based on their data, the critical point is approximately 100 mg acetone/L blood.
A principal finding in these studies is that the mode of acetone excretion appears to be
dose-related. At low concentrations acetone appears to be excreted primarily through expiration.
At concentrations above 15 ppm acetone appears in the urine at about 1% of the exposure level.
Levels of acetone lost through expiration increase disproportionately at higher concentrations.
21
3.5. PBPK MODELING
Information on the toxicokinetics of acetone and isopropanol, whose major metabolite is

acetone (Nordmann et al., 1973), has been used to develop physiologically-based,
pharmacokinetic models to compare the uptake, distribution and metabolism of the chemicals in
rats and humans by different routes of exposure (Clewell et al., 2001; Gentry, et al., 2002;
Kumagai and Matsunaga, 1995). The tissues described in the models include those associated
with uptake (lungs and skin), metabolism (liver) and fat storage with slowly- and rapidly
perfused compartments. The models have been validated for human exposure for the inhalation
pathway, but not the oral pathway.
4. HAZARD IDENTIFICATION
4.1. STUDIES IN HUMANS — EPIDEMIOLOGY AND CASE REPORTS
The available data on the human health effects of acetone exposures are limited. Most of
the available studies involve exposures in occupational settings.
4.1.1. Cancer Studies
Ott et al. (1983a,b) conducted a retrospective cohort study to evaluate the effects of
methylene chloride on the hematologic and circulatory systems of the workers at a plant that
manufactured cellulose diacetate and cellulose triacetate. The study monitored several causes of
death, including deaths from malignant neoplasms. The control cohort selected for the study was
a second plant that also manufactured cellulose triacetate fibers but used acetone as a solvent.
The two plants were in close proximity to each other and were operated by the same company.
The study involved production employees who worked in areas of high exposure to either
methylene chloride and acetone (as the exposed cohort) or just acetone (for the control cohort)
between January 1, 1954, and January 1, 1977. Employment ranged from three months to 23
years with time-weighted-average acetone concentrations of 380 to 1,070 ppm depending on the
job category. In the study, 948 acetone-exposed workers were the reference cohort for
comparison to workers exposed to acetone plus methylene chloride; comparisons to unexposed
controls were not made. For the acetone-exposed workers, the total number of deaths observed
from all causes was 24 and 3 for men and women, respectively, compared with the total expected
of 53.8 and 6.7 for men and women, respectively. There were no deaths among the 107 nonwhite
women in the exposed cohort. Among the acetone-exposed workers the incidence of “malignant
22
neoplasms” was 5 and 2 compared with an expected incidence of 10 and 2.3 for men and
women, respectively.
This study has several shortcomings that limit its use for assessing health effects in
humans. The focus was on deaths resulting from cardiovascular effects from exposure to
methylene chloride; other health effects related to acetone exposure were secondary. Workers
exposed only to acetone were selected as the referent cohort. The incidence of death was
compared with expected deaths calculated from U.S. death rates for white men, nonwhite men,
and white women. Also, the acetone-exposed cohort is smaller than the methylene chloride
cohort. Finally, the study lists deaths by malignant neoplasms without noting the cell type or
target organ.
4.1.2. Noncancer Studies
4.1.2.1. Controlled Studies
Groups of four men were exposed to 0, 200, 1,000, or 1,250 ppm acetone for 3 or 7.5
hours/day for 4 consecutive days on 4 successive weeks with one week of fluctuating
concentrations (750-1,250 ppm) for 3 days (Stewart et al., 1975). Two groups of 2-4 women
were exposed to 0 or 1,000 ppm for 3 or 7.5 hours/day on 4 consecutive days over the course of
one week. During the first week and on the first day of each week all subjects were exposed to
ambient air and measurements of relevant parameters were determined to establish baseline
conditions and concentrations, followed by four subsequent days of exposure to acetone at the
designated concentration. Male subjects were exposed to concentrations of 200, 1,000 or 1,250
ppm and fluctuating concentrations (750-1,250 ppm for 3 days) during weeks 2, 3, 4, and 6,
respectively. Female subjects were exposed to 1,000 ppm during week 5.
The concentration of acetone in the breath, blood, and urine was directly related to
exposure concentration, but decreased steadily postexposure. Odor intensity increased with
exposure concentration, but no exposure-related adverse subjective signs were reported. Clinical
chemistry analyses, hematological analyses, urinalyses, electroencephalograms,
electrocardiograms, and cognitive and pulmonary function tests remained normal and did not
vary from preexposure levels. No neurological abnormalities occurred and the modified
Romberg test and the heel-to-toe test remained normal. The visual evoked response
measurements were taken following exposures on the second and fourth days of exposure during
each week of exposure. Three of four males exposed to 1,250 ppm had a statistically significant
23
increase in total visual evoked response amplitude on individual days, but no consistent pattern
was observed. An early menstrual period was reported by three of four women after four days of
exposure to 1,000 ppm for 7.5 hours. The significance of a premature menstrual period was not
clear, and the study authors stated that additional research was needed. This study is
characterized as a six-week study by ATSDR (1994) and is used to derive a minimum risk level
for intermediate and chronic exposure. As presented in its protocols, this study was conducted
over a six-week period; however, the subjects were only exposed to acetone for 4 days/week for
four weeks (males) and four days for 1 week (females). The visual evoked response
measurements were made after the second and fourth days of exposure which constitutes a
relatively short exposure period.
A group of 11 male and 11 female volunteers were tested for neurobehavioral

performance before, during, and after a 4-hour exposure to 250 ppm acetone (Dick et al., 1988,
1989). Initial analyses of all behavioral tests indicated that sex differences did not exist, with the
exception of the profile of mood states (POMS). Dual task performance measurements showed
mild but statistically significant increased response times and false alarm percent both during
and postexposure. Measurements of postural sway were also slightly increased from acetone
exposure, but statistical significance was not reached. Only men had a statistically significant
drop on the anger hostility scale of the POMS test. No effects were found on visual vigilance,
choice reaction time, and memory scanning.
4.1.2.2. Cohort Studies
Satoh et al. (1996) examined the neurotoxic effects of acetone in 110 male workers at an
acetate fiber plant. A total of 67 nonexposed male workers at the same plant served as controls.
Mean worker age and length of acetone exposure were 37.6 and 14.9 years, respectively. Tests
used to assess narcosis included finger tapping, simple reaction time, and choice reaction time.
Memory tests consisted of the Benton visual retention test and forward and backward digit span.
Workers were classified into highly exposed (>500 ppm), moderately exposed (250-500 ppm),
and less exposed (<250 ppm) groups as determined by the acetone level in the breathing zone.
Acetone levels in alveolar air, urine, and blood were directly correlated with exposure levels,
indicating that an equilibrium is reached under continuous exposure resulting in absorption of
acetone into the cardiovascular system. During or after work symptoms of eye irritation, tearing,
acetone odor, and nausea were reported by 13.7-45.1% of exposed workers vs. 3.9-23.5% of
unexposed controls. Over the previous six months, heavy or faint feelings in the head, nausea,
and weight loss were reported by 23.6-25.8% of exposed workers vs. 2.9-9.8% of controls. The
24
symptoms showed a dose-response relationship. The authors speculated that they were probably
the result of peak exposure during a single day. No differences between exposed workers and
controls were observed on the Manifest Anxiety and Self-rating Depression scales or for
electrocardiogram, phagocytic activity, hematology, and clinical chemistry. A statistically
significant decrease in simple reaction time and digit span activity was observed among exposed
workers aged 30-44 years, but not in workers aged <30 or $45 years. The study authors
questioned whether the differences in only one age group were real or chance findings.
In a similar study, 71 factory workers with a mean age and length of exposure of 36 and
14 years, respectively, were evaluated for both central and peripheral nervous system effects
(Mitran et al., 1997). Exposure concentrations over an 8-hour shift ranged from 416 to 890 ppm
acetone. Mood disorders; irritability; memory difficulties; sleep disturbances; headache;
numbness of the hands or feet; eye and/or nose irritation; bone, joint, and/or muscle pain;
nausea; and abdominal pains were reported slightly more frequently in exposed workers as
compared with controls. However, the time during the work shift when the symptoms occurred
or were reported was not stated. Results of motor nerve conduction tests on the median, ulnar,
and peroneal nerves indicated statistically significant reductions in latency, amplitude, and/or
duration of both proximal and distal responses, but no consistent pattern of effect.
A statistically significantly reduction in the nerve conduction velocity of all three nerves was
observed in exposed workers as compared with controls. Statistically significant delayed
reaction times for the visual test and a lower mean distributive attention score were identified in
exposed workers when compared with the controls.
Although the results of the Mitran et al. (1997) study indicate a possible neurotoxic effect
from acetone exposure, the study has several shortcomings. For example, there is too little
information for a critical appraisal of the study design such as selection of controls, parameters
used for matching for age and other variables, experimental procedures, i.e., blind versus non-
blind determinations of neurotoxicity, and temperature control during neurological performance
procedures. Age-matching of the subjects and consistent temperature control are critical
parameters in nerve conduction velocity measurements. Some information regarding the
selection of matched controls was reported by Mitran (2000) in a reply to an analysis of the
Mitran et al. (1997) study made by Graham (2000). Additional confounding factors include: (1)
no dose-response relationship was established; and (2) it is not possible to rule out co-exposure
to other toxicants as the factory was a coin and metal plant and exposure to metals and other
contaminants may be likely. Some of these issues are discussed in an EPA memorandum by
Boyes and Herr (2002).
25
Eight occupationally-exposed and eight nonexposed workers were studied for subjective
symptoms on nine shift days (during work) and during leisure at 8 hours following termination
of exposure (Kiesswetter et al., 1994). The authors recorded the feeling of well-being at the
start, middle and end of each shift, and at eight hours following termination of exposure for nine
separate days. Personal measurements indicated that on average the workers were exposed to
2,730 mg/m3 (1,150 ppm) during the first half of the shift and to 1,720 mg/m3 (725 ppm) during
the second half of the shift. For each determination of well-being measurement the subjects
reported on tension, tiredness, complaints, and annoyance. The only endpoint demonstrating a
relationship between ambient acetone concentrations and ratings of well-being was in the
“annoyance” category which tended to occur during the middle and at the end of the shift.
Semen parameters were evaluated for 25 workers at a reinforced plastic production plant
(Jelnes, 1988). Breathing zone measurements indicated mean acetone concentrations of 69.6
(range of 21.9-246.4) ppm, 162 (55.7-562.9) ppm, and 94.5 (28.3-189.9) ppm approximately 28,
15, and 10 weeks, respectively, prior to semen analyses. Concurrent exposures to slightly higher
levels of styrene also occurred. Semen from the reference group was collected from samples
deposited at a fertility clinic. Semen volume, sperm concentration, and serum concentrations of
follicle stimulating and luteinizing hormones were not different from age-matched controls. The
percent of live sperm was significantly higher (p # 0.01; 80% vs. 68%) whereas the percent of
immobile sperm was significantly lower (p # 0.01; 30% vs. 40%) in the exposed group as
compared with controls. The exposed men also had a decrease in the percent of normal sperm
(47 vs. 60%) due to increased percentages of amorphous and pyriform sperm head shapes. This
study is limited by several factors, including the fact that the semen samples used in the control
group were collected from a fertility clinic, and subjects were concurrently exposed to styrene.
4.1.2.3. Case Reports
Clinical signs and symptoms of toxicity were reported for workers at three manufacturing
plants where acetone was used along with other solvents (Parmeggiani and Sassi, 1954; as
reviewed in OECD, 1998). Drowsiness, eye and throat irritation, dizziness, inebriation, and
headache were complaints from six employees exposed to 309-918 ppm for up to three hours
over a 7-15 year period. At the second plant where acetone concentrations ranged from 84 to
147 ppm, four workers reported nausea, abdominal pain, headache, vertigo, loss of appetite,
vomiting, and other debilitating symptoms. At the third site, eye, nose, throat, and bronchi
irritation, along with central nervous system disturbances were documented for 11 workers
exposed to 13-86 ppm acetone in conjunction with “high” concentrations of carbon disulfide.
26
Filter press operators were evaluated for clinical signs and symptoms of toxicity over a
two year period (Raleigh and McGee, 1972). During the process of cleaning presses, filter cloths
saturated with cellulose acetate dissolved in acetone were removed and replaced, which caused
short-term (about 2-3 hours) exposure to much higher acetone concentrations than normally
present in the work area. During the first year of the study, average acetone concentrations in
the breathing zone of workers while pulling filters was 2,300 ppm (326-5,548 ppm) and while
dressing presses was 300 ppm (69-904 ppm). In contrast, acetone concentrations in the general
air were 110 ppm (29-286 ppm). Similar concentrations were measured during the second year
of the study. Of the nine workers monitored, seven complained of eye irritation, four of throat
irritation, and two of nasal irritation, three reported headaches, and three noted lightheadedness.
Generally, the symptoms were transient, intermittent, and occurred when the concentrations of
acetone “greatly” exceeded 1,000 ppm. No indications of central nervous system effects were
found as assessed by gait, the finger-to-nose test, and Romberg sign (loss of joint position sense).
Effects from oral exposure to acetone are limited to case reports. In one case report, a
17-month-old girl was given approximately 4.88 mL/kg (3,850 mg/kg bolus dose) of acetone
through her gastronomy tube (Herman et al., 1997). The child was found gagging, unresponsive,
and diaphoretic with dilated sluggish pupils, right arm tonic-clonic activity, and left eye
deviation, and she was unresponsive to verbal or painful stimuli. Serum ketones were still
present at a 1:32 dilution and the abdomen was distended and firm. Following intubation and
supportive therapy, the child recovered fully. Another case report described a 53-year-old
woman admitted to the hospital after ingestion of nail polish remover (Ramu et al., 1978). Vital
signs were generally normal, but neurological examination showed that even though she was
oriented, the patient was lethargic but arousable and had a shortened attention span. Her blood
acetone concentration was 0.25 g/dL. The woman was admitted for observation and her
condition gradually improved as blood acetone levels declined.
27
4.2. PRECHRONIC AND CHRONIC STUDIES AND CANCER

BIOASSAYS IN ANIMALS
4.2.1. Prechronic Studies
4.2.1.1. Oral Studies
Groups of five male and five female F344/N rats were administered acetone in drinking
water at concentrations of 0, 5,000, 10,000, 20,000, 50,000, or 100,000 ppm for 14 days (NTP,
1991; Dietz et al., 1991). Time-weighted average doses for males were 0, 700, 1,600, 2,600,
4,300, and 6,900 mg/kg-day, respectively, and for females 0, 750, 1,500, 2,300, 4,350, and 8,600
mg/kg-day, respectively. All animals survived to termination. Water consumption was reduced
in both sexes at 50,000 and 100,000 ppm. Final body weights of males given 50,000 and
100,000 ppm were 87 and 63%, respectively, of controls, and final body weights of females
given 100,000 ppm were 87% of controls. Increases in relative liver and kidney weights were
observed in both sexes at $20,000 ppm, relative testes weights were increased at $50,000 ppm,
and thymus weights were decreased in “exposed” animals; further details of organ weight data
were not included. Bone marrow hypoplasia was noted in all high-dose males, but in none of the
controls. More detailed hematology and clinical chemistry studies were not included.
Groups of five male and five female B6C3F1 mice were administered acetone in drinking
water at concentrations of 0, 5,000, 10,000, 20,000, 50,000, or 100,000 ppm for 14 days (NTP,
1991; Dietz et al., 1991). Time-weighted average doses for males were 0, 960, 1,600, 3,900,
6,300, and 10,000 mg/kg-day, respectively, and for females 0, 1,600, 3,000, 5,500, 8,800, and
13,000 mg/kg-day, respectively. Water consumption was reduced in both sexes at 50,000 and
100,000 ppm and final body weights of high-dose males and females were slightly less than the
controls. All animals survived to termination. Kidney weights were increased at $50,000 ppm
for males and females, and liver weights were increased at $5,000 ppm in males and $20,000
ppm in females. Dose-related increases in the incidence and severity of centrilobular
hepatocellular hypertrophy were observed in males at $20,000 ppm and in females at $50,000
ppm. Hematology and clinical chemistry evaluations were not conducted.
Groups of 10 male and 10 female F344/N rats were administered acetone in the drinking
water at concentrations of 0, 2,500, 5,000, 10,000, 20,000, or 50,000 ppm for 13 weeks (NTP,
1991; Dietz et al., 1991). Time-weighted average doses for males were 0, 200, 400, 900, 1,700,
and 3,400 mg/kg-day, respectively, and for females 0, 300, 600, 1,200, 1,600, and 3,100 mg/kg
day, respectively. No deaths occurred in any group. Water consumption was decreased in high-
28
dose males and in females given 20,000 and 50,000 ppm acetone. Mean final body weight of the
high-dose males was 81% of the controls; body weights of the females were unaffected by
treatment. No clinical signs of toxicity or ophthalmic abnormalities were observed in any group.
At necropsy, statistically significant (p # 0.01 or 0.05) increases in the following organ weights
were noted: relative kidney weights were 114% of controls for 20,000 ppm females and 126 and
123% of controls for 50,000 ppm males and females, respectively; relative liver weights were
110 and 112% of controls for 20,000 ppm males and females, respectively, and 115 and 105% of
controls for 50,000 ppm males and females, respectively; and relative testis weights were 119%
of controls at 50,000 ppm. Liver weight changes were not associated with microscopic lesions
and were thought to result from enzyme induction. In high-dose males, depressed sperm
motility, caudal weight, epididymal weight and an increased incidence of abnormal sperm were
seen (data for testicular effects were given only for the 0, 2,500, 10,000, and 50,000 ppm groups;
see also Section 4.3.1.1). Males given the two highest concentrations of acetone had increases in
the incidence and severity of nephropathy, indicating early onset and enhanced progression of
the disease. In males given 0, 2,500, 5,000, 10,000, 20,000, or 50,000 ppm acetone, nephropathy
was observed in all treatment groups including the controls. As such, the incidence of
nephropathy rated as mild was taken as the indicator of toxicity. For the 0, 2,500, 5,000, 10,000,
20,000, and 50,000 ppm doses the incidence of minimal nephropathy was 5, 8, 8, 9, 1, and 1 (out
of 10 animals) and for mild nephropathy 1, 0, 0, 0, 9 and 9 (out of 10 animals), respectively. The
authors of the study identify the kidney effects as the most prominent chemically-related effect.
Nephropathy was not observed in females. Pigment deposition in the spleen was observed in
10/10 males in the 20,000 and 50,000 ppm groups compared with 0/10 controls.
Other endpoints were noted at 20,000 and 50,000 ppm doses of acetone, including
statistically significant (p # 0.01 or 0.05) changes in hematology in males. For the 20,000 and
50,000 ppm groups, leukocytes were 125 and 133% of controls, erythrocyte counts 92% and
90% of controls, reticulocyte counts 75 and 68% of controls, hemoglobin levels 97% of controls
in both groups, mean corpuscular hemoglobin was 102 and 108% of controls, and mean cell
volume was 105 and 109% of controls, respectively. Changes in red blood cell parameters of
20,000 and 50,000 ppm males were consistent with mild macrocytic normochromic anemia with
a depressed regenerative response. Mild leukocytosis was also observed in high-dose females,
but this single difference was not considered biologically significant. Clinical chemistry
parameters were not measured. In summary, the testis, kidney, and hematologic system were
identified by the study authors as target organs for male rats, with a LOAEL of 1,700 mg/kg-day
and a NOAEL of 900 mg/kg-day. A LOAEL for female rats was not identified.
29
Groups of 10 male and 10 female B6C3F1 mice were administered acetone in the
drinking water at concentrations of 0, 1,250 (males only), 2,500, 5,000, 10,000, 20,000, or
50,000 (females only) ppm for 13 weeks (NTP, 1991; Dietz et al., 1991). Time-weighted
average doses for males were 0, 380, 600, 1,400, 2,300, and 4,900 mg/kg-day, respectively, and
for females 0, 900, 2,000, 4,200, 5,900, and 11,000 mg/kg-day, respectively. No deaths occurred
and no clinical signs of toxicity were observed in any group. Water consumption was not
affected in males; however, dose-related decreases in water consumption were seen in all treated
females. Body weight and growth of the treated animals were not affected in either sex.
Hematology parameters, sperm morphology, and vaginal cytology were not affected by acetone
treatment. Organ weights from the treated males were similar to the controls. However,
statistically significant (p # 0.01 or 0.05) increases in high-dose female absolute and relative
liver weights were 113 and 110% of controls, respectively. Statistically significant (p # 0.05)
decreases in absolute and relative spleen weights were 89 and 88% of controls, respectively.
The only microscopic lesion seen in mice was centrilobular hepatocellular hypertrophy, observed
in two high-dose females and considered due to enzyme induction. Mild hepatic changes were
observed in males exposed to $20,000 ppm for 14 days (see above) but did not persist after 13
weeks of exposure, suggesting the development of tolerance toward acetone. In summary, the
liver was identified as the target organ in male and female mice. The reference to this effect as
an adverse effect is uncertain because the morphological changes may reflect induction of
enzymes rather than an untoward effect on the liver. Effects that were noted in the rat,
particularly males, were not evident with the mice. The LOAELs for males and females were
4,900 and 11,000 mg/kg-day, respectively, and the NOAELs were 2,300 and 5,900 mg/kg-day,
respectively. It should be noted that the LOAEL for male mice was selected by the study authors
on the basis of the transient findings in the 14-day study.
Groups of 30 male and female Sprague-Dawley rats were administered acetone by oral
gavage at doses of 0, 100, 500, or 2,500 mg/kg-day for 90 days; 10 animals/sex/group were
designated for interim sacrifice at 46-47 days (American Biogenics Corp., 1986). Survival, body
weights, food consumption, ophthalmology examinations, and gross necropsy findings were
similar between the treated and control groups. Clear salivation was observed between day 27
and study termination in a total of 21 males and 24 females at the high dose. Red blood cell
parameters (hemoglobin, hematocrit, mean cell volume, and/or mean cell hemoglobin) in the
high-dose groups increased in a statistically significant (p # 0.01 or 0.05) manner for males at
interim sacrifice and for males and females at final sacrifice. However, the study author did not
consider the magnitude of the increases to be biologically significant. One animal in the control
group, one in the low dose group, and four in the high dose group died prematurely; the deaths
30
were attributed to dosing errors. Differences in clinical chemistry parameters were not dose-
related and were not consistent over time or between sexes. Statistically significant (p # 0.01 or
0.05) increases in the absolute and/or relative liver and kidney weights were observed in the mid-
dose females and in the high-dose males and females when compared with their respective
controls. Relative (to brain and/or body weights) liver and kidney weights of the high-dose
males were 111-117% of the controls. Absolute kidney weights of mid-dose females were 110-
112% of controls and absolute and relative kidney weights of the high-dose females were 114-
118% and 111-123%, respectively, of control levels. Absolute and relative liver weights of mid-
dose females were 115 and 113%, respectively, and of high-dose females were 121 and 115-
125%, respectively, of the controls. Although nephropathy incidence rates were similar between
the treated and control groups, an increase in the severity of tubular degeneration of the kidneys
in mid- and high-dose males and females, and hyaline droplet accumulation in mid- and high-
dose males was observed. Statistical comparisons were not conducted for the increased severity
of the kidney effect. However, the nephropathy exhibited a dose-response with respect to the
numbers of animals affected. The numbers of male rats exhibiting tubular degeneration
characterized as mild or moderate (in comparison with minimal) were 0, 1, 9, and 17 out of 30
animals for the 0, 100, 500, and 2,500 mg/kg-day group, respectively. Based on organ weight
changes and kidney lesions in males and females, the LOAEL for this study is 500 mg/kg-day
and the NOAEL is 100 mg/kg-day.
4.2.1.2. Inhalation Studies
Male Sprague-Dawley rats were exposed to 19,000 ppm (45,106 mg/m3) acetone for 3
hours/day, 5 days/week, for 8 weeks (Bruckner and Peterson, 1981a). Although body weight
gains of the treated animals were slightly less than air-exposed controls, statistical significance
was not reached at any time. There were statistically significant (p # 0.01) decreases in the
kidney weights of treated animals when compared with the controls after 4 weeks of exposure,
but which returned to control levels after 8 weeks. Serum SGOT activities were not markedly
altered in treated animals at weeks 2, 4, and 8; however, LDH activity, and BUN and liver
triglyceride levels were not affected during the study. No microscopic lesions were observed in
the liver, brain, heart, and kidneys of the acetone-exposed animals. Females were not included
and no other concentrations of acetone were tested.
31
4.2.2. Chronic Studies
No studies on the chronic toxicity of acetone to laboratory animals are available.
4.2.3. Cancer Studies
No studies are available on the carcinogenicity of acetone to animals. Acetone has been
extensively used as a vehicle to dissolve test chemicals in dermal studies in mice (NTP, 1991,
1995, 1997; Ward et al., 1986; Zakova et al., 1985). Generally mice received one to two
applications per week for up to two years without an increase in neoplasia or any other toxic
response. However, no studies contained a naive control to provide a basis for comparison.
4.3. REPRODUCTIVE AND DEVELOPMENTAL STUDIES
4.3.1. Reproductive Studies
Ten male rats (Mol/Wis., SPF) were administered 0.5% acetone (5,000 ppm) in the
drinking water for six weeks and then bred to untreated females of the same strain (Larsen et al.,
1991). An additional group of 10 males was treated for six weeks followed by a 10-week
recovery period prior to breeding. Doses to the animals were not calculated. No effects were
seen in either group for number of pregnancies, number of fetuses/litter, testis weights, or testis
histopathology. However, when combined with 0.13-0.5% 2,5-hexanedione (2,5-HD), acetone
potentiated the testicular atrophy induced by 2,5-HD. The single concentration used in this
study prevents identification of a LOAEL or NOAEL.
In a previously discussed study, groups of 10 male rats were administered 0, 2,500,

5,000, 10,000, 20,000, or 50,000 ppm acetone in the drinking water for 13 weeks (NTP, 1991;
Dietz et al., 1991 [see Section 4.2.1.1]). Data for testicular effects were given only for the 0,
2,500, 10,000, and 50,000 ppm groups. The high-dose group (3,400 mg/kg-day) had: increased
relative testis weights (119% of controls), decreased caudal and right epididymal weights (71-
80% of controls, P<0.05 for both), depressed sperm motility (66.8% vs. 75.7% for controls,
P<0.05), and an increased incidence of abnormal sperm (3.42% vs. 0.68% for controls, P<0.05).
Data on the relative testis weights are difficult to interpret. Normally, reproductive toxicants
generate a decrease in testis weights. However, the change in relative weights of the testis may
be a result of overall decrease in body weight. There were no dose-related statistically
32
significant differences in absolute testis weight. The NTP (1991) study authors state that the
decrease in sperm motility at the highest dose was consistent with mild reproductive effects.
These effects
are characterized as consistent with mild toxic effects on spermatogenesis. The authors also
noted that there were no morphological or histological effects seen microscopically.
Data on the toxicity of acetone on male reproductive organs suggest that at high doses
there is a mild testicular effect, as indicated by diminished sperm motility and malformed sperm.
Larsen et al. (1991) indicate that at drinking water doses of 5,000 ppm there is no effect on male
reproductive capacity. At comparable doses in the NTP (1991) and Dietz et al., (1991) studies,
there were no detectable reproductive effects. It is unknown if the reduced sperm motility and
higher percentage with malformations noted at 50,000 ppm translates into impaired reproductive
ability.
Reproductive toxicity studies following the inhalation route are not available.
4.3.2. Developmental Studies
Developmental toxicity studies following the oral route of exposure are not available.
Presumed pregnant Sprague-Dawley rats (26-29/group) were exposed to 0, 440, 2,200, or

11,000 ppm acetone for 6 hours/day on gestation days 6-19 (Mast et al., 1988). Maternal body
weights of the high-concentration dams were statistically significantly (p # 0.05) less than
controls on gestation days 14, 17, and 20 and cumulative weight gain was reduced from gestation
days 14 until termination. No clinical signs of toxicity or maternal deaths were observed.
Treatment did not affect maternal liver and kidney weights, number of implantations, mean
percent of live pups/litter, mean percent of resorptions/litter, and fetal sex ratios (Table 2).
Statistically significant (p # 0.05; 84%-86% of control) reductions in fetal body weights from the
high-concentration group were observed when compared with controls (Table 3). Since the
study does not report food consumption, the reduction in maternal and fetal weight as a function
of reduced food consumption is not known. The incidence of fetal malformations was not
33
statistically significantly increased in any exposed group. However, the percent of litters
containing at least one pup with a malformation was 11.5% in the 11,000-ppm group (3/26
litters) compared with 3.8% for the control group (1/26 litters). Fetal malformations occurring at
a single incidence in the high-concentration group varied and included cleft sternum, ectopic
heart, major vessel malformation, edema, arrhinia, and microstomia; in addition, two fetuses
from one litter had a missing tail. Although there are several types of developmental effects
noted, there is no effect that predominates. Maternal plasma acetone levels measured 30 minutes
postexposure were increased: by 17 hours postexposure, acetone levels in the dams exposed to
440 and 2,200 ppm were similar to control levels, but remained elevated in the dams exposed to
11,000 ppm.
Table 2. Rat body, uterine, and extragestational weight gain of pregnant dams
Parameter Acetone Concentration (ppm)
0 ppm 440 ppm 2,200 ppm 11,000 ppm
Number of Dams 26 27 29 26
Gestational Day Maternal Weight (grams)
0 272.6±17.7 273.6±15.7 273.1±18.6 274.5±19.9
6 295.4±22.3 290.5±28.5 298.5±19.2 299.0±23.9
10 312.5±18.8 311.2±20.0 314.7±21.1 304.4±24.9
14 337.6±19.8 333.8±21.5 334.8±22.1 322.1±28.9
17 362.8±21.5 362.0±23.8 359.8±24.5 348.7±26.1
20 401.2±29.5 398.5±27.9 390.8±28.1 371.3±29.1*
Uterine Weight 83.2±19.0 79.7±14.5 74.5±16.3 67.1±13.9*

(grams)
Extra-gestational 45.3±16.7 45.1±13.2 43.2±15.2 29.7±14.4*

Weight Gain
(grams)
*P<0.05
34
Table 3. Average fetal weights in rats

0 ppm 440 ppm 2,200 ppm 11,000 ppm
Litters Examined 26 27 29 26
Fetal Weight 3.6±0.4 3.7±0.4 3.5±0.3 3.1±0.3*

(M&F)
Male Weight 3.7±0.4 3.8±0.2 3.6±0.3 3.1±0.3*

(grams)
Female Weight 3.5±0.3 3.6±0.2 3.4±0.3 3.0±0.3*

(grams)
Percent Male 51.6±13.7 49.9±14.4 51.9±15.5 49.5±13.2

Fetuses
* P<0.05
Presumed pregnant Swiss (CD-1) mice (26-31/group) were exposed to 0, 440, 2,200, or
6,600 ppm acetone for 6 hours/day on gestation days 6-17 (Mast et al., 1988). No clinical signs
of toxicity, maternal deaths, or effects on maternal body weights were observed (Table 4). Fetal
weights in the 6,600 ppm exposure group were about 8% lower than controls (1.3 g±0.1 and 1.2
g ±0.1, for control and 6,600 ppm groups, respectively, p #0.05) (Table 5). Statistically
significant increases in the absolute and relative liver weights of the 6,600 ppm group were
observed when compared with the controls, which may be indicative of enzyme induction.
Maternal kidney weights, number of implantations, mean percent of live pups/litter, percent of
total intrauterine deaths, and fetal sex ratios were not affected by treatment. A statistically
significant increase in the percent of late resorptions/litter was observed in the high-
concentration group as compared with the controls (7.8% vs. 3.2%, respectively). However, this
increase was not sufficient to result in a decrease in the number of live fetuses/litter. The
incidence of fetal malformations and variations in mice was not affected by exposure at any level
with the exception of a statistically significant increase in the percent of fetuses (on a litter basis)
with reduced ossification of the sternebrae in the high dose group (1.5 % ±4.0 and 9.9 % ±17.6
for controls and 6,600 ppm groups, respectively) (Table 6). The authors stated that this effect
might not be biologically significant since the incidence was <10%. No differences in the
percent of live fetuses per litter with at least one malformation were seen between the control
group (28.2% ±28.6) and the 6,600 ppm group (25.4% ±27.7) when all malformations were
combined.
35
Table 4. Mean body, uterine, and extragestational body weight gain of pregnant mice
0 ppm 440 ppm 2,200 ppm 6,600 ppm
Number of Dams 26 28 29 31
Gestation Day Maternal Body Weight (grams)
0 27.7±2.6 27.3±2.7 27.5±3.3 27.5±2.5
6 29.9±2.6 29.7±2.7 30.0±3.8 30.0±2.7
9 31.5±2.7 illegible 32.2±3.5 32.2±2.7
12 37.1±3.1 illegible 37.5±4.1 37.6±3.3
15 44.6±3.9 44.7±4.2 45.1±5.0 45.5±3.7
18 52.5±5.0 53.0±5.9 52.1±8.4 53.0±6.6
Uterine Weight (grams) 18.7±3.4 18.4±3.8 17.9±3.5 17.6±2.7
Extra-gestational 6.1±2.1 7.3±2.8 6.7±4.9 7.7±3.0

Weight Gain (grams)
Table 5. Average fetal mouse weight and percent male fetuses

0 ppm 440 ppm 2,200 ppm 6,600 ppm
Litters Examined 26 28 29 31
Fetuses Examined 292 307 318 344
Fetal Weight (grams) 1.3±0.1 1.4±0.1 1.3±0.1 1.2±0.1*
Male Fetuses (grams) 1.4±0.1 1.4±0.1 1.3±0.1 1.2±0.1*
Female Fetuses (grams) 1.3±0.1 1.3±0.1 1.3±0.1 1.2±0.1*

* P<0.05
36
Table 6. Mean percent of live fetal mice per litter with malformations and variations
Parameters Acetone Concentrations (ppm)
0 ppm 440 ppm 2,200 ppm 6,600 ppm
Malformation
Folded Retina 0.6±3.3 0 0 0.5±3.0
Exencephaly 0 0 0 0.4±2.2
Limb Flexure 1.3±3.9 0.6±3.4 0.5±2.7 0.2±1.4
Fused Ribs 0 0 0.3±1.9 0
Fused Sternebrae 0 0 0.3±1.5 0
Kinked Tail 0 0.3±1.5 0 0
Percent Fetuses/Litter with 1.6±4.1 0.9±3.7 1.1±3.5 0.9±2.9

$1 malformation
Other Variations
Supernumerary Ribs 23.3±27.3 29.5±28.0 20.7±21.8 12.9±17.1
Misaligned Sternebrae 5.1±10.0 8.2±12.2 5.0±7.7 9.4±14.5
Extra Sternebral 0.4±2.0 3.2±16.8 0.2±1.3 0

Ossification Sites
Dilated Ureter 0 0 0 1.2±4.6
Reduced Sternebral 1.5±4.0 2.4±4.9 2.7±6.6 9.9±17.6*

Ossifications
% Fetuses/Litter with $1 28.2±28.6 38.1±29.4 26.5±24.3 25.4±27.7

Variation or Reduced
Ossification
* P<0.05
4.4. OTHER STUDIES
4.4.1. Neurotoxicity Studies
Ladefoged et al. (1989) compared the effects of exposure to ethanol, acetone, and 2,5-
hexanedione alone or in combination for six weeks. From the third week on, mice were
monitored for nerve conduction velocity and rotarod performance. No effects on nerve
conduction velocity or on balance time with the rotarod test were observed in male Wistar rats
administered 0.5% acetone in the drinking water for six weeks. Peripheral distal axonopathy
37
was not observed in Sprague-Dawley rats (sex not specified) given 0.5% acetone for eight weeks
followed by 1% acetone for four weeks in the drinking water (Spencer et al., 1978). While water
consumption was reported in graphical form by Ladefoged et al. (1989), it was not reported by
Spencer et al. (1978) and the animal doses are unknown.
In a subchronic schedule-controlled operant behavior (SCOB) study-rats were trained to

press a lever to obtain food in standard behavioral test chambers (CMA, 1997). Operant sessions
were conducted 5 days/week for 9 weeks prior to acetone exposures. The operant set included
fixed ratio response rate, fixed ratio pause duration, fixed interval response rate, and fixed
interval index of curvature. Forty male Crl:CDBR rats (10 per group) were exposed to
concentrations of 0, 1,000, 2,000 or 4,000 ppm acetone for 6 hours/day, 5 days/week for 13
weeks. Of the four measures, only the mean fixed ratio pause duration demonstrated a response
at the two higher levels, starting with week eight and continuing through weeks 13 and 15 for the
two highest levels, respectively. The NOAEL and LOAEL for the mean fixed ratio pause
duration measure are 1,000 and 2,000 ppm, respectively. The author stated that the differences
observed were the result of unusual performance by the control group and were not treatment-
related. The control group exhibited an unexplained increase in the pause duration between week
seven and the end of the study. The other SCOB measures failed to demonstrate exposure-
related effects.
Acetone was evaluated for effects on a delayed match-to-sample task in male juvenile
baboons (Geller et al., 1979b). Animals were continuously exposed in inhalation chambers to
500 ppm for 7 days. Even though accuracy was not affected by exposure, two of four animals
showed marked changes in the number of extra responses made during exposure compared with
their own baseline; for one animal, extra responses increased at all test times, whereas for the
other, extra responses increased for the first two days of exposure and consistently decreased
thereafter.
Male rats were exposed by inhalation to 19,000 or 25,300 ppm acetone for 3 hours and
subjected to a series of performance/reflex tests both during and after exposure. Concentration-
dependent central nervous system depression was evident during exposure as measured by a
battery of simple tests of unconditioned performance and reflexes. Recovery was apparent 9
hours after exposure to 19,000 ppm and 21 hours after exposure to 25,300 ppm (Bruckner and
Peterson, 1981b).
38
Female CFE rats were exposed by inhalation to 3,000, 6,000, 12,000, or 16,000 ppm
acetone for 4 hours/day, 5 days/week, for 10 exposures (Goldberg et al., 1964). Growth rate was
not affected during the study. The two highest concentrations produced ataxia in several animals
after the first exposure, however, tolerance developed and this was not observed on subsequent
days. Avoidance behavior was inhibited at concentrations of 6,000 ppm and higher.
Male Sprague-Dawley rats, trained on a fixed ratio-fixed interval (FR-FI) schedule of

reinforcement, were exposed to 150 ppm acetone for 0.5, 1, 2, or 4 hours. No effects were
observed during the 0.5-hour exposure. Both FR and FI rates increased during the 1-hour
exposure and decreased during the 2-hour exposure as compared with controls. During the 4-
hour exposure, FI responses approximated control levels for two rats and were above the control
level for the third animal, whereas FR rates were below controls for two of the three animals
(Geller et al., 1979a).
4.4.2. Isopropanol Studies
Considering that isopropanol is metabolized mainly to acetone (Brugnone et al., 1983;

Slauter et al., 1994; Nordmann et al., 1973), toxicity studies on isopropanol may provide
additional information on the toxicity of acetone. The kinetics of acetone formation from
isopropanol varies among the available studies and depends on the species and concentration of
parent compound. The study descriptions for isopropanol, as indicated below, are not intended to
be an exhaustive review of the scientific literature but may inform toxicological issues raised by
data limitations for acetone.
Bevan et al. (1995) conducted a two-generation reproduction toxicity study on rats with
isopropanol. Thirty male and thirty female Sprague-Dawley rats (designated as the P1
generation) were dosed by oral gavage with 0, 100, 500, and 1,000 mg/kg-day for at least 10
weeks prior to mating. The females were dosed during mating, gestation and lactation while
males were dosed during mating and delivery. The second generation parental (P2) were
selected from the offspring of the first generation (F1) and were dosed for 10-13 weeks before
mating to produce a single litter.
Findings in the P1 group include increased liver weight in the P1 males and females, and
P2 females. The authors reported that no adverse effects of treatment were evident from the
39
gross postmortem examination. Histopathology revealed kidney effects in the mid- and high-
dose P1 males and in all treated groups of the P2 male rats. The effects were characterized by an
increased number of hyaline droplets in the epithelial cells of the proximal convoluted tubules,
increased incidence and severity of epithelial degeneration and hyperplasia, increased incidence
of proteinaceous casts in the renal tubules, and increased mononuclear infiltration. With the
exception of six male rats (out of 26) that exhibited centrilobular hypertrophy of the liver, there
were no other treatment-related microscopic changes observed in the reproductive organs, liver,
or kidneys, nor were gross abnormalities evident.
The authors reported that the F1 and F2 offspring that survived to scheduled termination
had no treatment-related abnormalities. There was a statistically significant decrease in survival
of the F1 offspring in the high dose group from postnatal day 0-4 for the high dose group and on
postnatal day 4 only for the 500 mg/kg-day group. A statistically significant decrease in survival
index was also seen in the F2 offspring in the mid and high dose groups on postnatal days 1 and
7. Statistically significant decreases in F1 male body weights from the high dose group were
found, when compared with controls on PND 0 and 1 and in male and female offspring in the F2
generation at PND 0, 1 and 4 compared with the controls. The lower fetal body weight may
reflect the larger litter sizes seen with the treatment (F1 12.4 pups/litter for controls vs. 14.4 in
the high dose group; F2 13.2 vs. 14.4 in the controls and high dose group, respectively). There
was an increased incidence of postweaning mortality in the high dose group of the F1 generation
which coincided with the onset of gavaging with isopropanol. The authors speculate that this
may be due to an immature metabolic system that is incapable of effectively metabolizing
isopropanol.
The only reproductive parameter that appeared to be affected by isopropanol treatment

was a statistically significant decrease in male mating index in the P2 high dose group (73.1%
vs. 93.3% for controls P<0.05). This is “slightly” below the historical control values for P2 male
mating index (range 80-96.6%, mean 91%) (study authors characterization). In interpreting this
study the authors questioned the significance of this finding: they noted no similar effect in the
P1 generation, no treatment-related effect in the P2 female fecundity index or fertility index, the
absence of adverse effects on litter size, and the lack of histopathological findings in the male
reproductive organs. The authors also note that all of the females that mated became pregnant.
Tyl et al. (1994) conducted developmental studies on rats and rabbits. In the rat study, 25
dams were dosed by gavage at levels of 0, 400, 800, and 1,200 mg/kg-day from gestation days 6
through 15. One dam in the mid-dose and two dams in the high-dose group died during
40
treatment. Maternal body weights were statistically equivalent across treatments but maternal
weight change was statistically reduced at the high dose group for the gestational period PND 0-
20 (141.8 vs. 157.8 for controls). The authors speculate that the difference may be due to the
reduced uterine weight. The fetal weights were reduced at the mid- (97.3, 94.7, and 94.3%) and
high- (92.1, 91.9, and 95.4%) dose group for all fetuses, male fetuses, and female fetuses,
respectively. There were no late fetal deaths observed in this study and there was no evidence of
treatment-related external, visceral, skeletal, or organ malformations or variations in the
offspring. The study authors reported that isopropanol, when administered by gavage to rats, is
maternally toxic at 800 and 1,200 mg/kg-day including lethality. The study authors note that
there is no evidence of teratogenicity at doses that are maternally toxic by the gavage route of
administration.
New Zealand white rabbits (15 per group) were dosed by gavage at doses of 0, 120, 240,
and 480 mg/kg-day from gestation day 6 through 18. Four does in the high dose group died
during the dosing period following dosing. The mortality was considered treatment-related.
Mean maternal body weight was lower in the high-dose group compared with those in the other
groups from the onset of dosing through delivery. A statistically significant reduction in
maternal body weight change was observed in the high dose group compared with other groups.
This coincided with lower food consumption in that group. The study authors reported that dams
in the high dose group exhibited flushed or warm ears early in the dosing. They noted that this is
indicative of typical adult mammalian alcohol intoxication.
There was a statistically significant reduction in mean fetal body weight per litter in the
high dose group for all fetuses (93.5%), male fetuses (92.7%), and female fetuses (86.0%)
compared with controls. There were no other statistically significant, treatment-related effects
on any gestational parameters. There were no treatment-related changes in the incidence of
external, visceral, skeletal, or total malformations or variations in any dose group. The authors
established maternal and developmental NOAELs of 240 and 480 mg/kg-day, respectively, for
the rabbits. Overall, the authors concluded that the only expression of toxicity is the reduction of
fetal weight and there were no signs of teratogenicity in either rats or rabbits. The study authors
concluded that isopropanol was not teratogenic.
Bates et al. (1994) conducted a developmental neurotoxicity study on Sprague-Dawley

rats. Four groups of 64 mated dams were administered isopropanol by gavage at doses of 0, 200,
700, and 1,200 mg/kg-day from gestation day 6 through postnatal day (PND) 21. On PND 4 the
litters were culled to 4:4 sex ratio, when possible, or 5:3. One male and female set of pups from
41
each litter was tested for motor activity using a figure-eight maze (PND 13, 17, 21, 47 and 58); a
second set was tested for auditory startle (PND 22 and 60); and a third set for learning and
memory in an active avoidance test (5 days at PND 60-64). One dam in the high dose group
died on PND 15. No other exposure-related clinical signs were observed in the dams. There
were no treatment-related effects on maternal or pup weight, sex ratio, pup developmental
landmarks or pup survival. None of the neurodevelopmental assays revealed any treatment-
related developmental effects in the offspring. The total motor activity increased to a maximum
on day 47 for all groups (including the control) which decreased for all groups in day 58
compared with day 47. The study authors state that this is a normal occurrence and, therefore,
does not constitute a treatment-related effect. Maximum startle amplitude and latency to
maximum amplitude were unaffected by isopropanol treatment. Similarly, examination of
learning and memory performance, as indicated by the active avoidance test, revealed no
treatment-related effects. The pathology assessment of the brains revealed no treatment related
abnormality in either the brain weights or histology. The authors note that a consistent finding in
both the developmental and neurodevelopmental studies (both studies were conducted at the
same facility) was the almost complete lack of clinical signs in the pups even at maternally toxic
(lethal) doses.
Burleigh-Flayer et al. (1994) conducted an inhalation neurotoxicity study on CD-1 mice

and Fischer rats. Five groups of rats or mice (10/sex) were exposed to isopropanol at
concentrations of 0, 100, 500, 1,500 and 5,000 ppm for 6 hours/day, 5 days/week for 13 weeks.
The study authors report that during exposure, some animals demonstrated narcosis, ataxia, and
hypoactivity. This effect was not seen in rats following the first week of the study. Female rats
demonstrated a 57% increase in motor activity in the 5,000 ppm group at weeks nine and 13.
Otherwise, there were no changes in any of the parameters of the functional observational
battery. No effects were seen with males. The authors report an increase in body weight and
body weight gain in female rats in the 5000 ppm group which corresponded with increased food
and/or water consumption.
In addition to neurotoxicity, the authors conducted hematological and histological

examinations of the animals. Hematological effects in male and female rats in the form of
reduced erythrocyte counts that were observed at six weeks in the high-exposure group were no
longer present at 13 weeks. In female mice at the high exposure, similar blood effects were
evident at 13 weeks. No blood effects were evident in male mice. The only organs affected at
42
the end of the 13-week exposure were the liver and kidney. The liver weights of male and
female rats were increased 5 and 8% in the high exposure group, respectively, compared with
controls. Ar necropsy, there were no gross lesions determined to be exposure related. The only
microscopic change observed was hyaline droplets within the kidneys of all male rats (including
controls). The size and frequency of the hyaline droplets were increased for the isopropanol
exposure groups compared to the control group. These differences were not clearly
concentration-related, although the changes were most pronounced in the high dose group.
Neuropathologic examination revealed no exposure-related lesions in the central or peripheral
nervous system of exposed rats. Thus, neurological effects were noted at the highest dose and
kidney changes were noted at all doses.
In a subchronic study, Burleigh-Flayer et al. (1998) conducted neurotoxicity evaluations

on CD-1 mice and Fisher F344 rats. The animals were exposed to concentrations of 0, 100, 500,
1,500, or 5,000 ppm of isopropanol for 6 hours/day and 5 days/week for 13 weeks. The study
authors reported ataxia, narcosis, hypoactivity, and/or a lack of a startle response following
exposure in some rats or mice in the 5,000 ppm group and in some mice in the 1,500 ppm group.
In the functional observational battery, none of the males exhibited statistically significant
effects compared with the control group while motor activity increased in the female group
following either 9- or 13-week exposures in the 5,000 ppm group. In a follow-up study, two
groups of 30 female Sprague-Dawley rats were exposed to concentrations of either 0 or 5,000
ppm of isopropanol. The two groups were divided into either 9- or 13-week sessions, and
spontaneous motor activity was assessed after 4, 7, 9, 11, and 13 weeks of exposure.
A statistically significant increase in motor activity was observed after 4 weeks of exposure
compared with the control group. The increased activity was not evident 2 days after cessation
of the 9-week exposure or 2 weeks after cessation of the 13-week exposure.
Nelson et al. (1988) conducted a developmental study on groups of 15 pregnant SD rats

exposed to isopropanol at concentrations of 0, 3,500, 7,000, or 10,000 ppm for 7 hours/day on
gestation days 1-19. The study authors report that at the two highest exposures, dams exhibited
signs of narcosis, an unsteady gait after exposure, a statistically significant reduction in food
consumption and reduced weight gain. With repeated exposure, the symptoms were less evident
indicating adaptation to isopropanol. Animals at the lowest exposure exhibited no abnormal
symptoms. Animals in the mid- and high-exposure group exhibited reduced food intake and
reduced weight gain during the first two weeks of the study at 7,000 ppm and for the entire study
at 10,000 ppm. Of the 15 dams in the high exposure group 6 animals were not pregnant, 4 had
total resorptions of the fetuses, and the remaining 5 dams had an increased number of resorptions
43
compared with controls. Offspring in the 7,000 and 10,000 ppm group had an increased
incidence of rudimentary cervical ribs. Overall, the incidence of litters with skeletal or visceral
malformations were 4/15, 1/14, 12/13, and 5/5 for the 0, 3,500, 7,000, and 10,000 ppm groups,
respectively. The malformations and variations seen at the higher concentrations in this study
were reported at maternally-toxic exposures. The only concentration at which maternal toxicity
was not observed was 3,500 ppm. At this exposure level, the only effect observed was a 3.8%
decrease in fetal weight in females (3.00 g ±0.38 vs. 3.12 g ±0.29 for controls) and a 4.3%
decrease in males (3.13 g ±0.36 vs. 3.27 g ±0.27 for controls).
4.4.3. Genotoxicity Studies
The genotoxicity of acetone has been well studied in in vitro assays, with the results
almost entirely negative (ATSDR, 1994; OECD, 1998; U.S. EPA, 1988b; WHO, 1998). All
studies cited in the GENE-TOX data base were negative, with the exception of one study for
which no conclusion was drawn.
Neither sister chromatid exchange (SCE) nor chromosome aberrations were induced in
Chinese hamster ovary cells by acetone at a concentration not exceeding 1% in the culture flask
with or without metabolic activation (Loveday et al., 1990). Acetone was also negative for
inducing sister chromatid exchanges in human (Tucker et al., 1993) and nonhuman (Latt et al.,
1981) cell types in the absence of metabolic activation. Acetone did not induce chromosome
aberrations in vitro (Preston et al., 1981).
Concentrations of acetone up to 0.6% did not change the background DNA synthesis
rate, i.e., induce unscheduled DNA synthesis, in cultured human epithelial cells. Higher
concentrations (up to 10%) inhibited background synthesis in a concentration-related manner
(Lake et al., 1978). The chemical was negative for reverse mutations at concentrations up to 10
mg/plate in the Ames reversion test with five strains of Salmonella typhimurium in the presence
or absence of a metabolic activation system (NTP, 1991; Kier et al., 1986; De Flora et al., 1984).
Cell transformation was not seen in Syrian hamster embryo cells at acetone concentrations up to
8% (Heidelberger et al., 1983). Acetone was not mutagenic to Arabidopsis at concentrations up
to 500 mM (Rédei, 1982). Male and female hamsters did not show an increase in micronuclei in
polychromatic erythrocytes in the bone marrow following injection with 865 mg/kg (Basler,
1986).
44
In contrast to the above reports, acetone, at concentrations of 6.98-7.83%, produced

aneuploidy in an inconsistent manner, but did not induce recombination or point mutation in
Saccharomyces cerevisiae. However, overnight storage on ice of cells in growth medium
containing acetone resulted in strong induction of aneuploidy (Zimmermann et al., 1985). The
significance of this study is unknown.
As an "-ketoaldehyde, methylglyoxal, a potential metabolite of acetone, is chemically

reactive in Schiff-base type reactions with amines or thiol groups. It has tested positive in some,
but not all, genotoxicity studies including S. typhimurium and E. coli reverse mutation assays
(Mitchell, 1993; Kranendonk et al., 1996), Chinese hamster lung cell HGPRT assays (Cajelli et
al., 1987), and SCE induction assays (Tucker et al., 1989; Nishi et al., 1989; Migliore et al.,
1990). It should be noted that no genotoxicity studies for acetone have included enzyme
activation systems specific for the production of methylglyoxal.
4.5. SYNTHESIS AND EVALUATION OF MAJOR NONCANCER EFFECTS

AND MODE OF ACTION — ORAL AND INHALATION
Potential exposure to acetone occurs through normal endogenous production and from
potential exogenous sources. Following the ingestion of food, glucose, starches, and other
carbohydrates are used to generate essential cellular components, energy, glycogen, or fatty
acids that may ultimately be stored as fat. Under normal conditions, the body uses glucose from
consumed foods and glycogen from the liver to provide energy. In a study designed to measure
the levels of acetone in the blood, urine and breath of occupationally-exposed and nonexposed
workers, Wang et al. (1994) reported a mean value of 840 :g/L of acetone in the blood of the
nonexposed cohort. Stewart et al. (1975) reported blood acetone concentrations in male subjects
ranging from 0.73 to 1.32 mg% and in females ranging from 2.86 to 3.58 mg% prior to initial
acetone exposure. Acetone blood concentrations in the range of 0.3 to 2.0 mg/100 ml are
reported to be normal in healthy individuals (Physicians Desk Reference, 1976). However, under
conditions of stress such as fasting and exertion, the metabolism of fat can result in higher levels
of acetone in the body. In particular, the brain and reticulocytes, which are incapable of using fat
as an energy source, rely on the regeneration of glucose, where acetone serves as an intermediate
in the metabolic process, as the energy source. As such, acetone is a normal cellular constituent
that the body is capable of metabolizing at low concentrations. Acetone also occurs
endogenously at elevated levels under some disease conditions, most prominently with diabetes.
The normal endogenous turnover rate (mg/kg-day) for acetone is not known.
45
Several studies in humans and animals suggest that acetone has a short persistence in the
bloodstream. Following exposure, acetone is rapidly absorbed and distributed throughout the
body and is rapidly lost. The ability to compensate for fluctuations in the concentration of
acetone is a function of both the chemical’s physicochemical properties and the body’s
biological capabilities, including: the absorption, metabolism, and excretion components of
toxicokinetics. Therefore, the body appears capable, under normal circumstances, of controlling
the concentration of acetone in the blood and tissues. At low concentrations, such as those
generated endogenously under normal conditions, acetone is largely retained and serves as an
intermediate in gluconeogenesis. Under high-level exposure conditions, excess acetone is
eliminated. Acetone has a high KB/A (blood-air partition coefficient), thereby suggesting a high
capacity for absorbing exogenous acetone vapors during inhalation while contributing to the
retention of acetone in the bloodstream. However, under high levels of exposure only about
45% of the inhaled acetone is actually dissolved in the bloodstream, a level considerably lower
than what is predicted based solely on the KB/A. The limited number of studies on the absorption
of acetone in humans via the gastrointestinal tract suggest that most ingested acetone is absorbed
into the body (Haggard et al., 1944; Price and Rittenberg, 1950).
Metabolism affords another mechanism for compensating fluctuating acetone levels in

the body. As proposed by Casazza et al. (1984) and based on several rodent studies, two
possible pathways for the metabolism of acetone have been characterized. The methylglyoxal
pathway leads to the formation of glucose and is mediated largely by enzymes from the hepatic
microsomes. A second pathway appears to be extrahepatic and proceeds through the formation
of 1,2-propanediol. Studies by Kosugi et al. (1986a,b) indicate that the methylglyoxal pathway
predominates at low acetone concentrations and serves as a mechanism for gluconeogenesis. At
higher concentrations acetone is also metabolized via the extrahepatic 1,2-propanediol pathway,
which is believed to occur largely in the kidney. This pathway may contribute to the production
of glucose, but could also be used as a means to readily eliminate acetone from the body.
A third mechanism that allows the body to adjust for elevated exposure to acetone is
excretion. The fate of acetone in the body appears to be dose- or concentration-dependent. At
low levels, human data indicate that acetone is mostly lost through metabolism to other cellular
constituents, whereas at higher levels, nonmetabolized acetone is detected in the expired air
(Wigaeus et al., 1981). Exposure to acetone at ambient air concentrations below 15 ppm
produces no detectable acetone in the urine (Kawai et al., 1992), whereas at higher
concentrations acetone is detected in urine at approximately 1% of the blood plasma
concentration (Wang et al., 1994; Wigaeus et al., 1981). At significantly higher concentrations,
some studies have found that the relative amount of acetone excreted increased relative to the
46
amount ingested, thereby increasing the rate at which acetone is lost from the body. As such, the
body maintains feedback systems that both actively and passively control the amount of acetone
taken up and retained.
Similarly, Reichard et al. (1979) working with fasting individuals, found that following
intravenous administration of 14C-acetone, the specific activity of 14C-acetone decreased while
the plasma concentration of acetone remained stable. Overall, the data indicate that excess
acetone may be readily lost from the body.
Based on laboratory studies and human data, there are primarily five toxicological
endpoints associated with exposure to acetone through inhalation or ingestion in humans and/or
rodents, including: eye and respiratory irritation, nephrotoxicity, hematological effects,
neurotoxicity, and male reproductive effects. Effects in humans via inhalation exposure have
been studied and generally appear during exposure to concentrations of 500 ppm or above (Dick
et al., 1988, 1989; Satoh et al., 1996; Mitran et al., 1997). Some of the effects are largely
subjective, including: eye and nose irritation, mood swings and, at higher levels, nausea. The
irritating effects of acetone may be due to its lipid solvent properties. Considering that the effects
appear during exposure and dissipate following cessation of exposure indicates that they are
caused by the parent material rather than a metabolite. Data on the neurotoxic effects of acetone
are limited to inhalation exposure. The neurotoxic effects noted in humans are reported to be
both mild and transient. Mitran et al. (1997) reported statistically significant reductions in nerve
conduction velocity in workers. Animal data are generally consistent with the effects noted in
humans. Other studies demonstrate transient impaired learning or response following inhalation
exposure in rodents (CMA, 1997). This is consistent with the relatively nonpolar, lipophilic
properties of acetone, which enhances its ability to cross the blood-brain barrier. As acetone is
lost from the system, either by excretion or metabolism, irritation and other neurotoxic effects
subside.
The most extensive animal studies conducted on acetone are drinking water and gavage
studies. Both rats and mice in the Dietz et al. (1991) and NTP (1991) studies had higher levels of
hemoglobin, lower red blood cell count (male rats only), and higher mean cell volume at the
highest concentrations. The conditions are symptomatic of megaloblastic anemia. In humans,
the condition is associated with folic acid and vitamin B12 deficiencies. In the 14-day study at
the highest concentration (100,000 ppm) the study authors (Dietz et al., 1991) reported bone
marrow hypoplasia which supports the concern that bone marrow could be a target organ for
acetone at high doses. A second possible explanation is toxicity associated with depletion of
GSH from elevated levels of methylglyoxal. Ankrah and Appiah-Opong (1999), in a mouse
47
drinking water study, found that perinatal exposure to methylglyoxal resulted in depletion of
GSH stores in red blood cells and lower ability to refract oxidative stress.
Nephrotoxicity was noted in both the gavage and drinking water studies. In both cases
the effects were mild; however, the effect also occurred at a lower dose and was more
pronounced in the gavage study. Also in the gavage study, American Biogenics Corp. (1986)
noted tubular degeneration, an effect that was not found in the drinking water study. A possible
explanation is related to the metabolism of the chemical. At lower concentrations acetone is
metabolized mainly through the hepatic pathway; at higher concentrations the extrahepatic
pathway becomes more significant. Bolus administration of acetone in the gavage study can be
expected to lead to an acute increase in the amount of acetone in the blood. At higher
concentrations, acetone can be expected to reach target organs including the kidney, causing the
noted effect at lower concentrations than that seen following administration of acetone via
drinking water. This is consistent with the interpretation offered by the authors of the NTP
(1991) study, who state that a possible mechanism of action for chronic progressive nephropathy
is based on the effect of higher concentrations of protein in the serum resulting in higher blood
flow and glomerular filtration rate, leading to the burdened glomeruli becoming sclerotic (NTP,
1991). This effect could be exacerbated by lower water consumption that was seen at the higher
acetone concentrations.
Male reproductive effects were also identified in the rodent studies, including: testicular
higher relative testis weight, reduced sperm motility, and more abnormal sperm at the higher
doses compared with controls (NTP, 1991; Dietz, 1991). The study authors note that the data are
difficult to interpret because testicular toxicants typically decrease testes weights and therefore,
the relative increase in weight may be more indicative of the reduced body weights in the treated
animal studies. Other statistically significant reproductive effects indicate that the effects are
minimal. An increase in abnormal sperm from <1 to 3.5%, and decrease in sperm motility from
75.7 to 66.8% were seen at the highest doses. Although studies concerning the distribution of
acetone indicate that radiolabeled material reaches the testis (Wigaeus, 1982), there is no
evidence demonstrating that the radiolabel present in the testis is due to acetone or a metabolite.
The one study reporting reproductive effects in humans is inconclusive (Jelnes, 1988). While
there are no two-generation reproductive toxicity studies for acetone, data from isopropanol,
which is metabolized primarily to acetone, may provide useful information. A two-generational
gavage study (Bevan et al., 1995) in rats indicates that the only reproductive effect was a
statistically significant reduction in the P2, but not P1, male mating index in the high dose group
(1,000 mg/kg-day) which the study authors characterized as slightly below historical controls.
48
Data on developmental effects resulting from oral exposure to acetone are not available
but may be informed by the results of an inhalation study on acetone and from oral exposure data
on isopropanol. Inhalation studies by Mast et al. (1988) reported an increased incidence in
skeletal malformations in rats and delayed skeletal development in mice at high exposures,
although the types of malformations are inconsistent. The most consistent finding with exposure
to acetone was decreased fetal weight at high exposure (86-92% of control at 6,600 ppm for
mice, and 84-86% of control at 11,000 ppm for rats). The developmental effects of isopropanol
have been studied by the oral route of exposure. Bevan et al. (1995) showed there was a decrease
in the survival of F1 offspring at high doses of isopropanol (1,000 mg/kg-day), a decrease in
body weight in high dose F1 male rats and F2 male and female rats, and no treatment-related
abnormalities. Tyl et al. (1994) showed maternal toxicity at 800 and 1,200 mg/kg-day in rats and
no evidence of developmental effects in offspring. The authors also reported decreases in fetal
body weight at 480 mg/kg-day in rabbits with no other developmental effects noted. Gavage
studies (Tyl et al., 1994) reported NOAELs of 400 and 480 mg/kg-day for rats and rabbits,
respectively, based on reduced fetal weight. No effects were observed at doses up to 1,200
mg/kg-day on postimplantation loss, litter size, sex ratio, or external, visceral, or skeletal
abnormalities for either model. Similarly, Bates et al. (1994) found no evidence of
neurobehavioral or morphological abnormalities from the prenatal administration of isopropanol
by gavage to pregnant rats.
4.6. WEIGHT-OF-EVIDENCE EVALUATION AND CANCER CHARACTERIZATION
In accordance with the Draft Revised Guidelines for Carcinogen Risk Assessment (U.S.
EPA, 1999) data are inadequate for an assessment of the human carcinogenic potential of
acetone. This weight-of-evidence determination is based on the availability of one human study
of limited utility, no chronic animal studies, and no additional information on structural
analogues with known carcinogenic potential. Acetone has tested negative in almost all
genotoxicity studies. The previous IRIS entry included a weight of evidence classification of
Group D - not classifiable as to human carcinogenicity - based on the Guidelines for
Carcinogenic Risk Assessment (U.S. EPA, 1986a).
Acetone has a long history of industrial use as a solvent. To date there are no
epidemiological studies demonstrating an association between exposure to acetone and increased
risk of cancer. The only human study available is an epidemiological study of workers in a
cellulose acetate plant where the workers were exposed to acetone concentrations of 380-1,070
ppm (time-weighted average) (Ott et al., 1983a,b). In this study, 948 workers served as the
reference cohort for a comparison to workers exposed to a mixture of methylene chloride and
49
acetone. For the acetone-exposed workers, the total number of deaths observed from all causes
was 24 and 3 for men and women, respectively, compared with the total expected of 53.8 for
men and 6.7 for women. Among the acetone-exposed workers the incidence of “malignant
neoplasms” was 5 and 2 compared with an expected incidence of 10 and 2.3 for men and
women, respectively. Limitations of this study are discussed in Section 4.1.1. Although a
chronic bioassay has not been conducted for oral or inhalation exposure routes, acetone has
frequently been used as a solvent or vehicle control to dissolve test chemicals in dermal studies
in animals (NTP, 1991, 1995, 1997) with no evidence of increased tumor incidence.
Nevertheless, without a naive control the ability to determine the background incidence of cancer
following these exposures is limited. Chronic and less than lifetime studies on methylglyoxal, a
potential metabolite of acetone, reported no signs of cancer in rats, although both studies are
limited by methodology and reporting (Fujita et al., 1986; Takahashi et al., 1989). In addition, a
cancer bioassay (Burleigh-Flayer et al., 1997) in F344 rats for isopropanol (which is metabolized
primarily to acetone) by the inhalation route of exposure indicated an increase in interstitial cell
adenomas of the testes in male rats. The adenomas were found at an unusually low incidence in
control rats in this study which may confound the results. There were no increased frequencies of
neoplastic lesions noted for female rats of male or female mice in any exposure group.
4.7. SUSCEPTIBLE POPULATIONS
4.7.1. Possible Childhood Susceptibility
Little information is available on the potential for increased susceptibility to acetone

exposure in children. Acute oral LD50 values for 14-day-old rats (16-50 g), young adult rats (80-
160 g), and older adult rats (300-470 g) were 5.6, 9.1, and 8.5 ml/kg, respectively, indicating
little age-related variability (Kimura et al., 1971).
There are no available developmental studies on acetone by the oral route of exposure.
However, two-generational studies on isopropanol, which is mainly metabolized to acetone, may
provide some information on the potential effects of acetone [Section 4.4.2.]. The administration
of isopropanol caused a reduced survival index in both F1 and F2 generations in the high-dose
groups and decreased fetal body weight in F1 males and F2 males and females through gestation
day 4 (Bevan et al., 1995). Lehman et al. (1945) conducted a drinking water study on
isopropanol at doses up to 2,500 ppm which also reported retarded weight in the F1 generation
during the first week with the effect diminishing over time. The authors reported no other
remarkable findings.
50
Developmental studies by the inhalation route of exposure indicate that reduced fetal
body weight in high-dose exposure groups is the most consistent effect seen in rats and mice
(Mast et al., 1988) [Section 4.3.2.2.].
No other studies are available which examine age-related differences in effects of acetone
exposure. In addition, the endogenous production rate of acetone in children under normal (or
any other) conditions is not known.
4.7.2. Possible Gender Differences
No studies indicating differences in the uptake or retention of acetone between men and
women are available. In a measure of subjective effects, Dick et al. (1988, 1989) reported that of
volunteers exposed to 250 ppm acetone vapor for 4 hours, only males exhibited a statistically
significant reduction in the anger hostility scale of the profile of mood states. In animal studies,
male rats exposed to 20,000 and 50,000 ppm acetone in drinking water exhibited an increase in
severity of progressive nephropathy compared with controls. A similar effect was not seen in
females (Dietz et al., 1991; NTP, 1991). In addition, male rats were more sensitive than female
rats to hematological, hepatic and renal effects (Dietz et al., 1991; NTP, 1991, American
Biogenics Corp., 1986).
4.7.3. Other Possible Susceptible Populations
The role of acetone in fasting individuals and diabetics is complicated and not well
understood (Reichard et al., 1979, 1986; Owens et al., 1982). Acetone is produced
endogenously; higher concentrations are produced during fasting due to an increase in fat
catabolism, which can result in ketosis (Reichard et al., 1979). This implies that individuals on
fasting diets who are also exposed to exogenous acetone will have a higher body burden of
acetone than nondieters exposed to the same amount of exogenous acetone, perhaps making
them more susceptible to any adverse effects. Individuals with impaired kidney function, most
notably diabetics, may also constitute a susceptible population. Data from Dietz et al. (1991) and
NTP (1991) indicate kidney effects in male rats in which members of all dosing groups,
including the control group, exhibited mild progressive nephropathy. The possibility that
acetone exposure may accelerate this condition cannot be ignored. In addition, diabetics may be
more susceptible to acetone exposure due to the production of the methylglyoxal metabolite
which may contribute to the formation of glycated reaction products that are a complication of
long-term diabetes. Methylglyoxal has also been shown to affect glucose tolerance indicating
potential concerns for individuals with diabetes (Ankrah and Appiah-Opong, 1999).
51
To the degree that acetone exposure may contribute to hematological or kidney effects,
individuals on high protein/low carbohydrate diets as well as diabetics may also comprise a
susceptible population due to an already elevated endogenous acetone level.
One of the most studied effects of acetone is the induction of microsomal enzymes,
particularly CYP2E1. Acetone thereby induces its own metabolism, and potentiates the toxicity
of numerous other chemicals (e.g., 1,1-dichloroethylene, 1,2-dichlorobenzene, carbon
tetrachloride) that are metabolized by CYP2E1 by enhancing their metabolism to reactive
intermediates (Morgott, 2001; Traiger and Plaa, 1974; Hewitt and Plaa, 1983; Brondeau et al.,
1989). The induction of CYP2E1 by acetone poses a potential concern for humans exposed to
acetone and those chemicals whose toxicity may be augmented by acetone.
5. DOSE-RESPONSE ASSESSMENTS
5.1. ORAL REFERENCE DOSE (RfD)
5.1.1. Choice of Principal Study and Critical Effect
There are no human studies or chronic animal studies available for the derivation of an
RfD. Two shorter duration studies are available, including a short-term exposure (14 days) study
using 5 animals per dose group (NTP, 1991; Dietz et al., 1991) that is not considered suitable for
the derivation of an RfD due to the acute nature of the dosing regimen. The principal study
identified for derivation of the oral RfD is a subchronic drinking water study (NTP, 1991; Dietz
et al., 1991). Acetone was administered in the drinking water (e.g., 0, 200, 400, 900, 1,700, and
3,400 mg/kg-day for male rats) of mice and rats for 13 weeks. Male rats appeared to be the most
sensitive species, with the kidney, hematologic system, and testes identified as target organs.
Enhanced progression of mild nephropathy and effects consistent with macrocytic
normochromic anemia with a depressed regenerative response were found at a high-dose of
1,700 mg/kg-day. In addition, depressed sperm motility, caudal and epididymal weights, and an
increased incidence of abnormal sperm occurred at 3,400 mg/kg-day. A LOAEL of 1,700
mg/kg-day and a NOAEL of 900 mg/kg-day were identified for mild nephropathy.
One additional 90-day study is available (American Biogenics Corp., 1986) in which rats
were administered 0, 100, 500, or 2,500 mg/kg-day acetone by gavage. Organ weight changes
and kidney lesions were identified at a dose of 500 mg/kg-day. While the gavage study included
clinical chemistry analyses, the data failed to show dose-related effects consistent with the
nephropathy noted in the histology, thereby raising questions about the significance of the effect.
Differences in the observed effect level in the drinking water study versus the gavage study may
52
relate to the method of administration. As noted earlier, acetone is readily absorbed through the
gastrointestinal tract (Section 3.1). Under conditions of short-term elevated exposure levels such
as those produced in gavage or bolus experiments, more acetone appears to be shunted to the
kidney, producing higher concentrations in the urine and higher rates of metabolism through the
propanediol pathway compared with the more gradual administration through drinking water.
This could account for differences in the nephropathy severity levels observed with drinking
water compared with gavage administration. For this reason, the gavage study was not chosen as
the principal study.
Mild nephropathy was chosen as the critical effect, and was seen in male rats only in the
Dietz et al. (1991) and NTP (1991) study. The choice of critical effect is supported by the report
of tubular degeneration of the kidneys in male and female rats and hyaline droplet accumulation
in males at 500 and 2,500 mg/kg-day in the American Biogenics Corp. (1986) study. An oral
gavage study on isopropanol (Bevan et al., 1995), which is metabolized primarily to acetone,
also supports nephropathy as the critical effect. In this two-generation reproductive toxicity
study, kidney effects (including an increased number of hyaline droplets in epithelial cells of the
proximal tubules and an increase in severity of epithelial degeneration) were noted at 500 and
1000 mg/kg-day P1 male rats and also at 100 mg/kg-day in P2 male rats. Changes in
hematological parameters (erythrocyte and leukocyte counts and hemoglobin levels) in male rats,
but not mice, were noted in the Dietz et al. (1991) study. Red blood cell parameters only were
significantly affected at 2,500 mg/kg-day in male rats in the American Biogenic Corp. (1986)
study.
EPA has examined general issues related to hyaline droplet formation and concluded that
a mode of action for renal pathology involving the formation of hyaline droplets containing "2u-
globulin is probably unique to male rats in certain instances and may not be relevant to human
risk assessment if certain criteria can be met (U.S. EPA, 1991b). In brief, these criteria include:
(1) an increase in the number and size of hyaline (protein) droplets in kidney proximal tubule
cells of treated male rats; (2) immunohistochemical evidence of "2u-globulin accumulating
protein in the hyaline drops; and (3) histopathological evidence of kidney lesions associated with
"2u-globulin nephropathology. In the NTP (1991) and Dietz et al.(1991) study in F344 rats, the
nephropathy is characterized by proximal tubule degeneration and intracytoplasmic droplets of
granules (hyaline droplets) in the proximal tubular epithelium with the severity of these lesions
showing a dose-related increase in males only. However, tubular degeneration of the kidneys in
mid- and high-dose males and females, and hyaline droplet accumulation in mid- and high-dose
males was observed in the American Biogenics (1986) study in SD rats. Furthermore, although
immunohistochemical analysis of hyaline droplet protein has not been performed following
53
acetone exposure, hyaline droplet histochemistry following inhalation exposure to isopropanol,

which is primarily metabolized to acetone, has been examined (Fowler and Martin, 1994). In this
study kidney sections were examined by immunohistochemical staining with "2u-globulin
following nine day inhalation exposure in F344 rats to 0, 5,000 and 10,000 ppm isopropanol.
There were no substantive differences between the exposure groups in intensity or distribution of
"2u-globulin. Appropriate studies to ascertain the issue of human relevance related to the
observed kidney nephropathy of acetone have not been performed. Thus, the above criteria have
not been met for acetone and the relevance of the observed kidney nephropathy remains
unknown. The renal lesions seen in both the drinking water (NTP, 1991; Dietz et al., 1991) and
gavage (American Biogenics, 1986) studies may represent an enhancement by acetone of the
chronic progressive nephropathy commonly seen in aging rats (Baylis and Corman, 1998).
5.1.2. Methods of Analysis
The data were analyzed using the NOAEL/LOAEL approach. Based on the findings of
NTP (1991) and Dietz et al. (1991), the critical effect is nephropathy. The LOAEL is 1,700
mg/kg-day (20,000 ppm dose group) and the NOAEL is 900 mg/kg-day (10,000 ppm dose
group) based on an increased incidence of mild nephropathy. The values were calculated as
time-weighted average doses based on body weights, water consumption, and nominal drinking
water concentrations. The available PBPK models for oral exposure were not used for the
derivation of the human equivalent dose because they have not been validated against human
data.
Minimal or mild nephropathy was present in all groups. The authors characterized mild
nephropathy as an adverse effect due to a progression of the condition with treatment. The
incidence and severity of nephropathy are provided in Table 7. One out of ten animals in the
control group was rated with a mild degree of nephropathy. No animals in groups dosed at
intermediate levels of 10,000 ppm or lower were rated with mild nephropathy. In contrast, nine
out of ten animals dosed at both 20,000 and 50,000 ppm were rated with mild nephropathy. This
type of response is not amenable to benchmark dose modeling, since a graded dose-response
curve is lacking (a response of 1/10 animals for controls compared with a response of 9/10
animals at the lowest response level) and the lowest response is much higher than what might be
considered a low, e.g., 10%, benchmark response. For this reason, the NOAEL/LOAEL
approach was used.
Table 7. Incidence and severity of nephropathy in F344 male rats following

drinking water administration of acetone
54
Severity of Control 2,500 ppm 5,000 ppm 10,000 ppm 20,000 ppm 50,000 ppm
Nephropathy
Minimal 5/10 8/10 8/10 9/10 1/10 1/10

Mild 1/10 0/10 0/10 0/10 9/10 9/10
5.1.3. RfD Derivation - Including Application of Uncertainty Factors (UFs)
The NOAEL used for RfD derivation is 900 mg/kg-day. The following UFs are applied
to the effect level: 10 for consideration of intraspecies variation (UFH; human variability), 3
(101/2) for interspecies differences (UFA; animal to human), 3 to account for extrapolation from
subchronic studies to chronic exposure conditions (UFs; subchronic to chronic), and 10 to
account for a deficient data base (UFD). The total UF = 10 × 101/2 × 101/2 × 10 = 1000.
An UF of 10 was applied for intraspecies uncertainty to account for susceptible

subpopulations. This factor accounts for humans who may be more susceptible to acetone
exposure than the general population but for whom data are not available. This may include
individuals who have elevated levels of endogenous acetone due to high-fat low-carbohydrate
diets, fasting conditions, or uncontrolled diabetes. In addition, the production of glycated
endproducts may be increased due to acetone exposure. These endproducts have been shown to
be responsible for many of the complications associated with diabetes.
An UF of 3 was used to account for laboratory animal-to-human interspecies differences.

This UF accounts for differences in the toxicokinetics and toxicodynamics between the model
species and humans. The data indicate that the toxicokinetics in the rat and humans are similar
and that both species eliminate acetone from the body efficiently (Haggard et al., 1944; Sakami,
1950; Sakami and Lafaye, 1951; Stewart et al., 1975; Reichard et al., 1979; Casazza et al., 1984;
Wigaeus et al., 1981; Kosugi et al., 1986a; Wang et al., 1994). In both humans and rodents
metabolism proceeds by a hepatic pathway at low concentrations and by an extrahepatic pathway
followed by excretion at higher concentrations but qualitative toxicokinetic comparisons
between rats and humans are not available (Haggard et al., 1944; Casazza et al., 1984; Wigaeus
et al, 1981; Kosugi et al., 1986b; Gavino et al., 1987; Kawai et al., 1992). Thus, the
toxicokinetic component of the UF for interspecies extrapolation is 3. The critical effects
identified from the principal study are kidney-related (Dietz et al., 1991; NTP, 1991). Male rats
given the two highest doses of acetone had increases in the incidence and severity of
nephropathy; the severity rating increased from minimal at low doses to mild at high doses. It is
not known with certainty whether the observed nephropathy is a result of male rat-specific
55
hyaline droplet formation due to "2u-globulin accumulating protein (see Section 5.1.1).
However, nephropathy was observed in all treatment groups including controls. The authors
report that the effects are morphologically similar to the spontaneously occurring and long-term
progression of nephropathy (chronic progressive nephropathy) found among aging rats. Acetone
exposure may serve to enhance this effect. However, the nephropathy was increased only from a
severity rating of “1, minimal” to “2, mild” on a scale of 1 to 5. For these reasons, the
toxicodynamic component of the UF for interspecies extrapolation is 1 indicating that humans
are not anticipated to be more susceptible than animals to the nephrotoxic effects of acetone
exposure.
An UF of 3 was used to account for extrapolation from subchronic studies to chronic

exposure conditions. The principal study is a subchronic study. No chronic studies are available.
However, acetone is endogenously produced in the human body. Several reports note the
presence of acetone in normal nonfasting individuals indicating that humans are routinely
exposed to acetone (Stewart et al., 1975; Physicians Desk Reference, 1976; Wang et al., 1994).
Toxicokinetic studies in both humans and animals indicate that acetone elimination occurs
through excretion, exhalation and metabolism by various routes.
In addition, acetone does not accumulate in the body nor are its metabolites considered
significantly toxic. Acetone is metabolized to acetol which in turn, is metabolized via two
potential pathways to glucose. Intermediates include methylglyoxal, 1,2-propanediol,
lactaldehyde, and lactate, none which have been demonstrated to be overtly toxic.
An UF of 10 was used to account for data base uncertainty. The available data base for
acetone includes subchronic gavage and drinking water studies in mice and rats, including
measurements of several reproductive parameters. There is one neurotoxicity study in rats which
evaluated effects on nerve conduction velocity and rotarod performance and one reproductive
toxicity study with a single dose regimen. The data base lacks a multigenerational study and
adequate studies of the oral neurotoxicity, developmental and developmental neurotoxicity of
acetone.
The only available neurotoxicity study in mice demonstrates no effects on nerve

conductivity and rotarod performance. Nevertheless, human inhalation studies on acetone, while
inadequate, indicate potential neurotoxic effects. This raises concern for neurodevelopmental
effects from oral exposure because the nervous system undergoes developmental processes
unique to early life stages.
56
There are no developmental toxicity studies for acetone by the oral route of exposure;
however, information on this endpoint is possibly informed by inhalation studies and studies on
isopropanol. Inhalation studies on acetone reported a slight increase in the incidence of skeletal
malformations in rats, although the types of malformations did not demonstrate a consistent
effect. The most consistent finding was decreased fetal weight that was not associated with any
other observable adverse effect (Mast et al., 1988). Developmental toxicity studies on
isopropanol following gavage administration to rats and rabbits indicated reduced fetal weight at
doses of 800 and 1200 mg/kg-day but no other effects at any dose (Tyl et al., 1994). In addition,
a two-generation gavage study on isopropanol in rats (Bevan et al., 1995) indicated a statistically
significant reduction in the P2, but not P1, male mating index at 1,000 mg/kg-day that the study
authors characterized as slightly below historical controls. On the other hand, a developmental
neurotoxicity study (Bates et al., 1994) on isopropanol in rats indicated no toxicity at doses as
high as 1,200 mg/kg-day. It is difficult to draw firm conclusions on the potential developmental
effects from oral exposure to acetone based on inhalation studies following acetone exposure and
oral studies using isopropanol. However, evaluated collectively, the data may indicate potential
developmental and reproductive effects resulting from ingestion of acetone.
The RfD is based on a NOAEL which obviates the need for an UF due to LOAEL to
NOAEL extrapolation.
A modifying factor was not needed.
RfD = NOAEL ÷ UF
900 mg/kg-day ÷ 1000 = 0.9 mg/kg-day
The RfD is applied to ingested acetone only and is in addition to the acetone formed
endogenously by catabolism of fat. The turnover rate (mg/kg-day) for the endogenous production
of acetone under normal conditions is not known. The RfD generated in this assessment differs
from the previous RfD (0.1 mg/kg-day). This difference is accounted for, in part, by a change in
the principal study. The previous RfD is based on the gavage study conducted by American
Biogenics Corp. (1986). The administered doses were 0, 100, 500, or 2,500 mg/kg-day. The
critical effect noted was kidney pathology, and the NOAEL was 100 mg/kg-day. The RfD
invoked uncertainty values of 10 for intraspecies and interspecies extrapolation, and 10 for
extrapolation from a subchronic to a chronic exposure scenario. Although the point of departure
noted in the gavage study is lower, the drinking water route more closely mimics potential long-
term human exposure scenarios and the study is considered more thorough.
5.2. INHALATION REFERENCE CONCENTRATION (RfC)

57
Several controlled and cohort studies from inhalation exposure to acetone are available.
Dick et al. (1988) reported effects from short-term exposure to 250 ppm acetone including a mild
statistically significant increase in dual task performance measurements and in the identification
of false alarms during and immediately following exposure. Other neurological measurements
demonstrated no treatment-related effects. In a cohort study of workers occupationally exposed
to acetone at levels exceeding 500 ppm, Satoh et al. (1996) found an increase in reports of eye
irritation and nausea compared with a nonexposed control cohort. The authors speculate that the
reports may be the result of peak exposures during the course of the day. They found no
differences between the exposed and nonexposed cohort for the Manifest Anxiety scale or the
Self-rating Depression scale. The only reported neurological effect was a statistically significant
decrease in simple reaction time and digit span among workers aged 30-44. No differences were
reported in workers in younger and older age groups. The study authors questioned whether the
differences in only one age group were chance findings. Kiesswetter et al. (1994) report in a
study of workers exposed to acetone concentrations of 725 and 1,150 ppm during the morning
and afternoon shifts, respectively, that the only category of well-being related to exposure of
acetone was in the “annoyance” category.
In a cohort study, 71 factory workers with mean age and length of exposure of 36 and 14
years, respectively, were evaluated for both central and peripheral nervous system effects
(Mitran et al., 1997). Exposure concentrations over an 8-hour shift ranged from 416 to 890 ppm
acetone. Mood disorders, irritability, memory difficulties, sleep disturbances, headache,
numbness of the
hands or feet, eye and/or nose irritation, bone, joint and/or muscle pain, nausea, and abdominal
pain were reported slightly more frequently in exposed workers as compared with controls. The
time during the work shift when the symptoms occurred or were reported was not stated.
Although the results of motor nerve conduction tests on the median, ulnar and peroneal nerves
indicated statistically significant reductions in latency, amplitude and/or duration of both
proximal and distal responses, no consistent pattern of effect was observed. Statistically
significant reductions in nerve conduction velocity in all nerves studied was reported in exposed
workers as compared with controls. For the exposed workers, statistically significant delays in
reaction time were observed for the visual test and a lower mean distributive attention score
when compared with the controls.
The Mitran et al. (1997) study presents minimal study information which confounds a
meaningful appraisal of the study design. This includes a lack of information regarding the
selection of controls, parameters used for age-matching and other variables, experimental
procedures, i.e., blind versus nonblind determinations, and temperature control. Age-matching
and consistent temperature control are known critical parameters in nerve conduction velocity
58
measurements. Additional potential confounding issues associated with this study include no
establishment of a dose-response relationship, and an inability to rule out coexposure to other
toxins as the factory was a coin and metal plant where exposure to other toxins might be
considered likely. Some of these issues are discussed in Boyes and Herr (2002). For these
reasons, the study is not considered appropriate for the establishment of an RfC for acetone.
Stewart et al. (1975) exposed 20 adults of both sexes to acetone vapor concentrations of
0, 200, 1,000, 1,250 ppm or varying concentrations (males) and 0 or 1,000 ppm (females) for
either 3½ or 7 hours for 4 days/week on successive weeks. One of four subjects exposed to
1,000 ppm and two of four subjects exposed to 1,250 ppm for 7 hours demonstrated a
statistically significant effect on the visual evoked response. The effect was reported following
either two or four successive days of exposure. The exposure regime indicates that the exposure
duration at each concentration did not extend beyond 4 days for each dose with a 3 day interim
period without dosing. While this study may be appropriate for establishing exposure limits for
short-term exposure, it is not suitable for deriving a reference value for chronic exposure.
Mast et al. (1988) conducted developmental studies in rats and mice by the inhalation
route of exposure. Statistically significant decreases in maternal weight and weight gain in rats
were observed at the highest (11,000 ppm) exposure when compared with the controls. The
effect was not observed in mice. The authors reported that there were no overt signs of
developmental toxicity in either rats or mice. There were single incidences of fetal abnormalities
in the high-exposure rats. Statistically significant changes in the incidence of fetal
malformations in mice were not observed following exposure at any level with the exception of
an increase in the percent of fetuses (on a litter basis) with reduced ossification of the sternebrae.
The authors stated that this might not be biologically significant since the incidence was <10%.
The offspring of rats and mice had a small, but statistically significant increase in late term
resorptions compared with controls but the increase was not considered to be enough to account
for a reduction in live fetuses. Similarly, there was a small but statistically significant decrease
in fetal weight of offspring in both rats and mice as described by the study authors. The
significance of the fetal body weight effect is questioned in light of the minimal severity of the
effect, and the negative findings of other parameters including resorptions, number of live births,
and number of births per litter, which were comparable to controls.
Overall, the most pronounced effect of acetone reported in human inhalation studies is
irritation of the eyes and respiratory tract. Additionally, human data indicate that exposure to
acetone may produce neurobehavioral effects. Studies that report responses over time note that
the most pronounced effects occur during initial exposure and dissipate over time. Similarly,
59
animal data also indicate that neurological effects from less than lifetime inhalation exposure are
mild and transient. Although the available data base may be sufficient to support concerns for
short-term exposure (based largely on irritation) extrapolation to chronic exposure is not
recommended. Available human and animal studies on acetone exposure provide insufficient
information for the generation of an RfC. The available PBPK models are not amenable for
route-to-route extrapolation from the oral route to the inhalation route of exposure since the oral
exposure models have not been validated.
More research is needed on the toxicity of acetone following inhalation exposure. It

should be noted that the previous IRIS assessment for acetone did not derive an RfC. Additional
information can be found in Sections 4.1 and 4.2.1.2 of this review.
5.3. CANCER ASSESSMENT
Studies necessary to support a quantitative assessment of the carcinogenicity of acetone

in humans are not available. One epidemiological study published in the literature considers the
incidence of cancer in occupationally exposed individuals (Ott et al., 1983a,b; and as reviewed in
ATSDR, 1994); however, this study has several limitations: acetone-exposed individuals served
as the referent cohort and not the study cohort, the study was limited in size, and cancer was not
the principal focus of the study. There are no available lifetime cancer studies on animals via
either the inhalation or ingestion route. Acetone has been used as a vehicle for dissolving test
chemicals for several dermal studies in animals with no observed tumor incidences (NTP 1991,
1995, 1997); however, in the absence of a naive control the data are not adequate to evaluate the
potential for the induction of cancer. There are no cancer studies in which low molecular weight
saturated ketones that are similar in structure to acetone are shown to be carcinogenic. In vitro
genotoxicity studies with acetone are almost uniformly negative.
Neither IARC nor NTP have a human cancer classification for acetone.
6. MAJOR CONCLUSIONS IN THE CHARACTERIZATION

OF HAZARD AND DOSE RESPONSE
6.1. HAZARD POTENTIAL
Acetone is used industrially as a solvent and feedstock; biologically it serves a role in

human metabolism during fasting/starvation. Commercial acetone is used in the production of
high-volume chemicals including methacrylates, bisphenol A and other ketones, and as a
solvent. Small amounts are used in the pharmaceutical industry. In humans, acetone is formed
60
endogenously under conditions of starvation, uncontrolled diabetes, or with high fat/low

carbohydrate diets. Under conditions where glucose stores are depleted acetone provides a
means of supplying glucose to tissues that are incapable of metabolizing fatty acids.
Exogenous acetone is readily absorbed via inhalation, ingestion, and dermal exposures.
The water solubility of acetone allows for broad distribution to the water compartments of the
body. At relatively low levels of exposure acetone may be lost through expired air and
metabolized through the methylglyoxal pathway. At higher levels a second pathway producing
1,2-propanediol becomes more active and nonmetabolized acetone is lost through the urine.
Earlier work on acetone metabolism proposed a metabolic pathway that produced an “active”
form of acetate and formate. Although the active form of acetate may be acetyl CoA, the
evidence to support the production of formate from acetone is sparse.
No human studies following oral exposure to acetone are available. Studies on rodent
exposure to orally-administered acetone have identified several treatment-related health effects.
Subchronic oral exposure resulted in kidney, testis, and hematologic system effects; however,
the effects were characterized as mild. Although the nephrotoxic effects noted in rodents have
been identified as the most critical effects, they tend to occur in male rats only and at high levels
of exposure (20,000 and 50,000 ppm in drinking water).
Inhalation studies in humans have been conducted on both volunteers and

occupationally-exposed individuals (Dick et al., 1988,1989; Kiesswetter et al., 1994; Stewart et
al., 1975). These studies have examined, almost exclusively, either the toxicokinetics or
neurological effects of acetone. The effects reported in these studies appear to be mild and
transient. Clinical studies and case reports suggest slight neurological effects, mostly of the
subjective type, in individuals exposed to varying concentrations of acetone. In most studies the
subjects report discomfort, irritation of the eyes and respiratory passages, mood swings, and
nausea following exposure to acetone vapor at concentrations of 500 ppm or higher. The fact
that the effects subside following termination of exposure indicates that acetone may be the
active compound, rather than a metabolite. Clinical chemistry analyses conducted in several
studies demonstrated no exposure-related effects. Data on nerve conductivity are inconclusive.
Case reports of accidental poisoning also indicate that the effects (e.g., lethargy and drowsiness)
are short-lived. Transient effects are seen in male rats exposed to acetone by inhalation.
Inhalation exposure to pregnant rats and mice did not cause statistically significant
malformations in the offspring, but did result in lower fetal body weights in both species.
There is one human epidemiological study for cancer incidence available that has several
61
limitations. There are no chronic animal studies that address the issue of the human carcinogenic
potential of acetone.
6.2. DOSE RESPONSE
Quantitative estimates of human risk as a result of oral exposure to acetone are based on
laboratory animal exposures because no human data are available. The reference dose for
chronic oral exposure is 0.9 mg/kg-day. The RfD is based on a subchronic oral study in the rat
(NTP, 1991; Dietz et al., 1991) with a NOAEL of 900 mg/kg-day and a composite UF of 1,000.
Although complete histopathological analyses were performed (both sexes were used, and both
mice and rats were studied) confidence in the study is medium because the study is a subchronic
rather than a chronic study. Confidence in the overall data base is medium because there are two
well-conducted subchronic studies and considerable supporting data on humans, but no chronic
or multigenerational oral studies on acetone are available. Additional information on the toxicity
of acetone is provided by studies on isopropanol, a chemical that is metabolized primarily to
acetone.
A reference concentration (RfC) has not been determined. There are no studies
demonstrating conclusive effects either in humans or animals arising from chronic exposure.
Confidence in the overall data base is low because little supporting data and no chronic or
reproductive inhalation studies are available.
Lack of sufficient evaluation of reproductive toxicity by either oral or inhalation

exposure is considered a data base deficiency. Changes in testicular weight were observed in
male rats following oral exposure and a premature menstrual period occurred in 3 of 4 women
acutely exposed by inhalation. The significance of these endpoints of reproductive toxicity in
men and women is unknown at this time.
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APPENDIX A. SUMMARY OF EXTERNAL PEER REVIEW

AND PUBLIC COMMENTS AND DISPOSITION
The support document and IRIS summary for acetone have undergone both internal peer review
by scientists within EPA and a more formal external peer review performed by scientists in
accordance with EPA guidance on peer review (U.S. EPA, 1998b, 2000a). Comments made by
internal reviewers were addressed prior to submitting the documents for external peer review and
are not part of this appendix. The external peer reviewers were tasked with providing written
answers to general questions on the overall assessment and on chemical-specific questions in
areas of scientific controversy or uncertainty. Comments were also received from the public. A
summary of significant comments made by the external reviewers and the public and EPA’s
response to these comments follows.
Comments from External Peer Review
1. Overall quality of the assessment.
Comment: The reviewers stated that overall the assessment is well conducted and well written
and that an adequate effort was made to integrate the findings of different toxicology studies.
One reviewer indicated that the animal studies were better characterized than the human studies.
Response: The human studies have been revised and more extensively characterized in Section
4.1.
72
2. The use of secondary data in the assessment.
Comment: The reviewers stated that the assessment appropriately uses secondary data and that
the studies are well characterized. Several reviewers recommended the inclusion of several
omitted studies and indicated that PBPK models for isopropanol and acetone (Gentry et al.,
2002; the abstract for this publication was cited by the reviewer) are available.
Response: The recommended studies for inclusion in the assessment were considered. The
available PBPK models are now included in the text in Section 3.5.
3. RfD derivation (selection of the principal study, critical effect, and generation of the
dose-response).
The external review draft assessment identified the Dietz et al. (1991) and NTP (1991)
subchronic drinking water study as the principal study. Kidney, hematological and testicular
effects were identified as the critical effects and a NOAEL/LOAEL approach was used for the
point of departure. The draft assessment used a NOAEL of 900 mg/kg-day (based on the time-
weighted average) for nephrotoxicity and applied a total UF of 3000. The data were determined
to not be amenable to benchmark dose analysis due to poor model fit. The individual UFs in the
draft assessment were applied as follows: intraspecies UF=10, interspecies UF=3, subchronic to
chronic UF=10, data base UF=10.
(A) Comment: All reviewers indicated the drinking water study is the best choice for the
principal study. All reviewers agreed that nephrotoxicity is the best available chemically-related
response to use as the critical effect although all stated that the significance of the effect and its
relevance to humans is questionable in that the effect was male rat-specific and may be due to
"2u-globulin-induced hyaline droplet formation. In addition, the change was similar to a
naturally-occurring nephropathy seen in aging F344 rats. Thus, at best this effect might represent
an earlier onset of a condition that would have occurred anyway. One reviewer recommended a
stronger rationale for the decision to apply a NOAEL/LOAEL point of departure in lieu of a
benchmark dose based on modeling. One reviewer indicated that toxicity information on acetone
may be available from studies on isopropanol, which when metabolized produces acetone.
Response: The renal effects chosen for the critical effect have been re-examined and significant
changes to the text have been made. The study descriptions in Section 4.2.1.1 have been
augmented with more detailed information on the observed effects. In addition, significant
changes have been made in Section 5.1.1 to justify and clarify the use of nephropathy as the
73
critical effect, particularly in reference to EPA Guidelines on Alpha2u-globulin: Association with

Chemically-Induced Renal Toxicity and Neoplasms in the Male Rat (U.S. EPA, 1991b). The
rationale for using a NOAEL/LOAEL approach in lieu of benchmark dose modeling has been
augmented in Section 5.1.2. In brief, the type of response observed is not amenable to
benchmark dose modeling since a graded dose response curve is lacking (a response of 1/10
animals for controls compared with a response of 9/10 animals at the lowest response level).
References to attempts at benchmark dose modeling have been omitted. A review of the data
base for isopropanol toxicity for oral and inhalation exposures has been included in Section
4.4.2. The data base has been taken into consideration in the acetone assessment, primarily with
respect to the application of UFs as indicated in Section 5.1.3.
(B) Comment: Two reviewers commented on the choice of UFs for the derivation of the RfD.
The comments were mainly directed at the use of a 10-fold UF for extrapolation from a
subchronic to chronic exposure duration. One reviewer questioned whether environmental
exposures to acetone at the level of the proposed RfD (0.3 mg/kg-day in the external peer
review draft) would present a negligible increase in the body burden compared with the acetone
that a healthy individual generates on a daily basis. This reviewer indicated that humans may
have an ability to equilibrate and metabolize excess acetone. Along these same lines, another
reviewer indicated that the pharmacokinetic behavior of acetone indicates that it does not
accumulate and is eliminated efficiently through metabolism and via partitioning into expired air
and urine. This reviewer stated that accumulation would not occur even with chronic daily
exposure to high levels. At physiological levels of acetone, the prevalent elimination pathway
would be through intermediary metabolism as indicated in Figure 1 of the Toxicological Review.
In addition, the statement was made that there appeared to be little difference in acetone doses
that were adverse in the two week study versus the 13 week study by NTP (1991). This reviewer
also questioned the use of a 10-fold UF for data base uncertainty given that inhalation studies
indicated little or no developmental hazard. The reviewer stated that the UF for data base
insufficiency could be reduced by supplementing the data base with the developmental studies
available in the inhalation data base.
Response: The UF for subchronic to chronic extrapolation has been changed from a 10 to a 3.
The text in Section 5.1.3 has been significantly augmented to reflect the rationale for this change.
The change is primarily a result of the comments of the external peer reviewers.
An UF of 10 has been retained to account for data base uncertainty. The available data base for
acetone includes subchronic gavage and drinking water studies in mice and rats, including
measurements of several reproductive parameters. There is one neurotoxicity study in rats which
74
evaluated effects on nerve conduction velocity and rotarod performance and one reproductive
toxicity study with a single dose regimen. The data base lacks a multigenerational study and
adequate studies of the oral neurotoxicity, developmental and developmental neurotoxicity of
acetone. The only available neurotoxicity study in mice demonstrates no effects on nerve
conductivity and rotarod performance. Nevertheless, human inhalation studies on acetone, while
inadequate, indicate potential neurotoxic effects. This raises concern for neurodevelopmental
effects from oral exposure because the nervous system undergoes developmental processes
unique to early life stages. It is difficult to draw firm conclusions on the potential developmental
effects from oral exposure to acetone based on inhalation studies following acetone exposure and
oral studies using isopropanol. The available inhalation studies on acetone and the oral and
inhalation data base for isopropanol (which is metabolized primarily to acetone) has been
included in the text and taken into consideration to inform the oral data base for acetone.
4. The decision to not derive an RfC.
The draft IRIS assessment of acetone does not derive an RfC.
Comment: All reviewers agreed with the decision to not derive an RfC. One reviewer indicated
it would be useful to augment the discussion of why the available studies are not adequate to
support an RfC. One reviewer stated that there may be enough toxicokinetic data available to do
a route-to-route extrapolation for oral to inhalation exposure. Another reviewer indicated that an
examination of potential route-to-route extrapolation should be discussed but that this endeavor
would still be insufficient to derive an RfC.
Response: Additional text has been added to Sections 4.2.1.2, 4.3.1.2, 4.3.2.2, 4.4.1.2 and 5.2 to
better describe the results of the available inhalation studies and the rationale for not deriving an
RfC. In addition, text has been added (Section 3.5) to indicate that the available PBPK models
have not been validated in humans for oral exposure making route-to-route extrapolation tenable.
5. Cancer weight of evidence determination.
The cancer weight of evidence determination in the external review draft assessment was based
on the Guidelines for Carcinogen Risk Assessment (1986) and the Proposed Guidelines for
Carcinogen Risk Assessment (1996). Since the release of the external review draft assessment,
EPA has begun to use the Draft Revised Guidelines for Carcinogen Risk Assessment (1999).
The 1996 and 1999 guidelines apply different descriptors for weight of evidence. The external
review draft classifies acetone as category D - not classifiable as to human carcinogenicity and
75
uses the weight of evidence descriptor that human carcinogenic potential cannot be determined
based on the existing (1986) and proposed (1996) guidelines, respectively.
Comment: The reviewers agreed with the weight of evidence classification and descriptor. One
reviewer recommended including classifications from other agencies, e.g., IARC, NTP.
Response: The weight of evidence classification according to the 1986 guidelines that was
included in the external review draft has been removed. Under the Draft Revised Guidelines for
Carcinogen Risk Assessment (1999), the weight of evidence descriptor for acetone has been
changed to indicate that “data are inadequate for an assessment of human carcinogenic
potential.”
The assessment has been revised (see Section 5.3) to reflect that IARC and NTP have not
assessed the human carcinogenicity of acetone.
6. Additional comments.
(A) Comment: All reviewers made editorial comments.
Response: Editorial comments have been taken into consideration and edits to the text have been
made in numerous instances.
(B) Comment: One reviewer noted that information from other agencies such as ATSDR,
NIOSH and WHO should be included in the assessment.
Response: Information used in the assessment of acetone by various agencies has now been
taken into account. The studies that were used in the evaluation by ATSDR, NIOSH and WHO
have been discussed in the text of the Toxicological Review. For example, the studies used by
ATSDR in their quantitation of inhalation toxicity (e.g., Stewart et al., 1975) have been
described in detail. The shortcomings of this study for the purposes of deriving a chronic RfC
have been described in Sections 4.1.2.1 and 5.2. WHO (1998) derived an RfD for acetone based
on the same principal study and point of departure (900 mg/kg-day) as that used in this
assessment. The WHO applied an UF of 100 (no explanation of individual uncertainty factors)
for an RfD of 9 mg/kg-day.
Comments from the Public
(A) Comment: One reviewer indicated there is very little discussion in the draft document of
normal levels of acetone found in healthy individuals as a result of endogenous production. The
76
reviewer pointed out that the document emphasized endogenous production of acetone only
under conditions of stress (i.e., diabetes or fasting conditions). The comparison of endogenous
levels under normal conditions with the RfD was stated to be an important consideration. The
reviewer indicated that a published study (Reichard et al., 1979) suggests that in normal healthy
adults the rate of acetone production is in the range of 41 mg/kg-day which can be doubled by
merely fasting. The reviewer indicated that the Reichard et al. (1979) study did not derive this
value directly but that it had been derived in Patty’s Toxicology (cited as Morgott (2001) in this
document). This background production would be more than 135-fold the RfD (0.3 mg/kg-day in
the external review draft). The reviewer stated that this information needs to be considered and
the apparent discrepancy between the wide range of normal endogenous production in healthy
adults and the proposed RfD needs to be reconciled.
Response: It is agreed that if the endogenous production of acetone (i.e., turnover rate expressed
as mg/kg-day) under normal conditions were known, it would be useful to provide comparisons
between this value and the RfD. However, the proposal to extrapolate this value from the
Reichard et al. (1979) study does not seem feasible. In this study acetone turnover was
monitored in lean and obese subjects under conditions of starvation ketosis (3 and 21 day
starvation periods). Nonfasting individuals were not monitored and acetone turnover rates under
normal conditions were not determined. The proposed derivation (Morgott, 2001) from the
Reichard et al. (1979) data utilizes the y-intercept of a graphical representation showing the
relationship between acetone turnover rates, expressed as micromoles per square meter per
minute, and the plasma acetone concentration from the data corresponding to individuals
undergoing starvation ketosis. The y-intercept (20 µmol/min-m2) is then converted to the
turnover rate of 41 mg/kg-day by utilizing the conversion factors of 70 kg body weight and 1.73
m2 surface area. It is not evident that extrapolation from data on individuals in starvation ketosis
would yield the acetone turnover rate in normal individuals. No other studies were located in
which the acetone turnover rate could be derived.
(B) Comment: One reviewer indicated the 10-fold UF for subchronic to chronic extrapolation
was not necessary. The reviewer stated that acetone exhibits low subchronic toxicity and that
there is evidence of acetone’s ready metabolism and elimination from the body. This reviewer
also stated that the 10-fold UF for data base insufficiencies also needs to be re-examined in light
of the existence of a negative oral one-generation reproduction study on acetone and the fact that
only minimal effects on selected male reproductive organs in other subchronic studies have been
reported. In addition, inhalation developmental toxicity studies in two species have been
conducted and resulted in high NOAELs.
77
Response: The UF for subchronic to chronic extrapolation has been changed from a 10 to a 3.
The text in Section 5.1.3 has been significantly augmented to reflect the rationale for this change.
The UF for insufficiencies in the data base remains at 10 (see response to Comment 3B above).
(C) Comment: One reviewer contended that EPA should state expressly that acetone is unlikely
to cause cancer in humans. The reviewer stressed that available information on acetone supports
a weight of evidence classification of “E” (evidence of noncarcinogenicity for humans) by
applying the Guidelines for Carcinogen Risk Assessment (1986) and “not likely to be
carcinogenic to humans” according to the Proposed Guidelines for Carcinogen Risk Assessment
(1996).
Response: The cancer weight of evidence determination is based on the lack of formal studies
that specifically address the potential for human cancer from exposure to acetone. In addition, in
determining the weight of evidence descriptor for acetone, EPA has considered such factors as
the absence of data on low molecular weight ketones causing cancer in humans or laboratory
animals; no identified mode of action data to support concern for the carcinogenicity of acetone;
routine use of acetone as a vehicle in dermal cancer studies (albeit in the absence of naive
controls); and the genotoxicity data which are largely negative. A search of the scientific
literature identified one human study that assessed cancer incidence and no chronic animal
studies. The limitations of the available studies are discussed in the Toxicological Review. The
absence of studies limits the ability to reach an affirmative determination of evidence of
noncarcinogenicity for humans.
Since the release of the external draft Toxicological Review and IRIS Summary for acetone,
EPA has begun to apply the Draft Revised Guidelines for Carcinogen Risk Assessment (1999) in
lieu of the Proposed Guidelines for Carcinogen Risk Assessment (1996) and the 1986 Cancer
Guidelines. Section 2.6.2 of the 1999 guidelines identify criteria for use of descriptors for weight
of evidence. The guidance states that application of the descriptor “not likely to be carcinogenic
to humans . . . is used when the available data are considered robust for deciding that there is no
basis for human hazard concern.” Examples provided in the guidelines concerning the strength
of data include extensive human experience that demonstrates a lack of carcinogenic effect, or
animal evidence that demonstrates lack of carcinogenic effect in at least two well-designed, well-
conducted studies. For these reasons, data are inadequate for an assessment of the human
carcinogenic potential of acetone using the Draft Revised Guidelines for Carcinogenic Risk
Assessment (1999).
78

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EPA/635/R-03/004

(CAS No. 67-64-1)

In Support of Summary Information on the Integrated Risk

U.S. Environmental Protection Agency

AUTHORS, CONTRIBUTORS, AND REVIEWERS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vi

2. CHEMICAL AND PHYSICAL INFORMATION

3.1.1. Oral Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4

3.1.2. Inhalation Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5

3.1.3. Dermal Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7

3.5. PBPK MODELING . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22

4.1. STUDIES IN HUMANS — EPIDEMIOLOGY AND CASE REPORTS . . . . . . . 22

4.1.1. Cancer Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22

4.1.2. Noncancer Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23

4.1.2.1. Controlled Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23

4.1.2.2. Cohort Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24

4.1.2.3. Case Reports . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26

4.2. PRECHRONIC AND CHRONIC STUDIES AND CANCER

4.2.1. Prechronic Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28

4.2.1.1. Oral Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28

4.2.1.2. Inhalation Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31

4.2.2. Chronic Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32

4.2.3. Cancer Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32

4.3. REPRODUCTIVE AND DEVELOPMENTAL STUDIES . . . . . . . . . . . . . . . . . . 32

4.3.1. Reproductive Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32

4.3.1.1. Oral Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32

4.3.1.2. Inhalation Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33

4.3.2. Developmental Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33

4.3.2.1. Oral Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33

4.3.2.2. Inhalation Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33

4.4. OTHER STUDIES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37

4.4.1. Neurotoxicity Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37

4.4.1.1. Oral Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37

4.4.1.2. Inhalation Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38

4.4.2.1. Oral Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39

4.4.2.2. Inhalation Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42

4.4.3. Genotoxicity Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44

4.5. SYNTHESIS AND EVALUATION OF MAJOR NONCANCER EFFECTS

AND MODE OF ACTION — ORAL AND INHALATION . . . . . . . . . . . . . . . 45

4.6. WEIGHT-OF-EVIDENCE EVALUATION AND CANCER

4.7. SUSCEPTIBLE POPULATIONS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50

4.7.1. Possible Childhood Susceptibility . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50

4.7.2. Possible Gender Differences . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51

4.7.3. Other Possible Susceptible Populations . . . . . . . . . . . . . . . . . . . . . . . . . . 51

5.1. ORAL REFERENCE DOSE (RfD) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52

5.1.1. Choice of Principal Study and Critical Effect . . . . . . . . . . . . . . . . . . . . . 52

5.1.2. Methods of Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54

5.1.3. RfD Derivation - Including Application of Uncertainty Factors (UFs) . . 55

5.2. INHALATION REFERENCE CONCENTRATION (RfC) . . . . . . . . . . . . . . . . . . 57

6. MAJOR CONCLUSIONS IN THE CHARACTERIZATION

OF HAZARD AND DOSE RESPONSE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60

APPENDIX A. SUMMARY OF EXTERNAL PEER REVIEW

AND PUBLIC COMMENTS AND DISPOSITION . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72

Lynn Flowers, Ph.D., DABT

U.S. Environmental Protection Agency

National Center for Environmental Assessment

Michael W. Broder, Ph.D.

U.S. Environmental Protection Agency

National Center for Environmental Assessment

Lynn Flowers, Ph.D., DABT

U.S. Environmental Protection Agency