Seblework Final PHD Dissertation 22aug2016

Human Exposure and Consumer Risk Assessment to Pesticide Use in Ethiopia
Seblework Mekonen Shegen
Dissertation in the fulfillment of the requirements for degree of Doctor (PhD) in Applied
Biological Sciences
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Promoters:
Prof. dr. ir. Pieter Spanoghe
Department of Crop Protection, Ghent University, Belgium
Dr. Argaw Ambelu
Department of Environmental Health Science and Technology, Jimma University, Ethiopia
Dean: Prof. dr. ir. Marc Van Meirvenne
Rector: Prof. dr. Anne De Paepe
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Members of the Jury:
Prof. dr. Ann Dumoulin (Chairman)
Department of Industrial Biological Sciences, Ghent University
Prof. dr. ir. Marijke D’haese (Secretary)
Department of Agricultural Economics, Ghent University
Prof. dr. ir. Patrick Van Damme
Department of Plant Production, Ghent University
Prof. dr. ir. Liesbeth Jacxsens
Department of Food Safety and Food Quality, Ghent University
Prof. dr. Luc Hens
Department of Human Ecology, Vrije Universitiet Brussel
3
Dutch translation of the title: Humane blootstelling en consumenten risicoanalyse van het gebruik van
bestrijdingsmiddelen in Ethiopië
Cover Photo taken and designed by author
Cited as Mekonen S. Human exposure and consumer risk assessment to pesticide use
in Ethiopia, PhD dissertation, Ghent University, 2016
ISBN 978-90-5989-911-7
Printing Universitypress,Zelzate:[email protected]
Copy right © 2016 Seblework Mekonen Shegen
The author and the promoters give the authorization to consult and to copy part of this work for personal
use only. Every other use is subject to copy right laws. Permission to reproduce any material contained in
this work should be obtained from the author.
4
Acknowledgments
Acknowledgments
i
Acknowledgments
ii
Acknowledgments
First of all, I would like to thank and glorify to the almighty God, for taking care of me all the
time and giving me health, rigor and courage to embark on the PhD work. I would like to express
my deepest gratitude to my promoter Prof.dr.ir. Pieter Spanoghe, at first for his willingness to
accept my request to pursue my PhD in his laboratory. His guidance, inputs and encouragement
have been inspirational in shaping my PhD dissertation. I am very grateful for his devotion to
correct and comment all the manuscripts I prepared for publication and the whole PhD thesis.
I am so grateful to the technical staff in the Laboratory of Crop Protection Chemistry, Lilian
Goeteyn and Claudine Schollaert for their support during the experimental work. Lilian, you
were very helpful to me in all aspects of academic, social and administrative processes. It is
unforgettable how you helped me to socialize in the lab and around Ghent. I am also very
grateful to Michael Houbraken and David Senaeve who were assisting me during instrumental
analysis of my samples in the laboratory. I am also indebted to Elvina Mortier, Caroline, Davina,
Kim, Wumbie, Chara, Edlbis and dr. ir. Bjorn Vandekerkhove for their support during my study
and all the things we share in the coffee room.
I also like to extend my gratitude to em. Prof. Walter Steurbaut and his wife Rita, for outings
around Ghent, Ostende, Brussels, and for the delicious dinner we had together, and overall
support to make me feel at home during my stay in Belgium. Prof. Steurbaut did support in
framing my PhD study during the inception of the PhD ideas.
I would like to forward special appreciation to my home institution, Jimma University, for giving
me a leave and other supports to undertake my study. I am grateful to Mrs. Zinash Solomon, an
administrator to the College of Health Sciences, for facilitating administrative issues that are
related to my study. The staff in the department of Environmental Health Science & Technology
were very cooperative. I would like to thank dr. Seid, Taffre, Asgdom, Higemengist, Embiale,
Dessalegn, Samuel, Wuhib, Hailu, dr. Alemayehu, dr. Taddesse, dr Abebe, dr. Dante, Tebkew,
Alebel, Gudina, Tesfalem, Tizita, Seyoum, Yared, Tenagne and Abeba, for the appreciations and
moral support.
I am deeply grateful to Ghent University for the full financial support through BOF, a special
research fund, which made this PhD study become realized.
iii
Acknowledgments
Without the motivation, support and care of my family, this PhD study would not come true. I
would like to extend my deepest gratitude to my beloved husband dr. Argaw Ambelu. Arg, your
support, encouragement, love, care and guidance were very helpful for me throughout the period
of my PhD study. I remember how much you were carful for our children by acting as a father
and a mother in all my absence from home. My love, your usual dream and effort, to make me
one of the strongest women, is always motivating me to move forward. Arg, I really do not have
enough words to thank you. I understand how much my lovely children, my son Fsihatsion and
my daughter Sosna Argaw, missed me during my stay in Belgium. My children, your eagerness
to see my final defense and to hear your mother being called ‘doctor’, gave me the energy and
additional encouragement to accomplish this work. I am also very much thankful to my sisters,
Hamelmal Mekonen, Lidia and Aselefech who were taking care of my children during my stay in
Belgium.
My heartfelt appreciation and gratitude also goes to my father Mekonen Shegen and my mother
Masho Terefe, who were my base to reach this stage. The support and encouragement of my
brothers, Amdemariam and Ayele, and my sisters, Tsehay, Abaynesh, Genet, Zebenay,
Yeshihareg, Aynalem, and Kidist were very helpful to me to reach this level.
My appreciation also goes to the staff of OBSG, i.e. Marleen, Annemie, Father Charles and
Isabel for their support during my stay in Ghent. Marleen, thank you so much for reserving a
room to me every year and making me comfortable. I also would like to thank all those people
whom I have forgotten to mention, but who directly or indirectly contributed to the successful
accomplishment of this work.
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Table of contents
Table of contents
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Table of contents
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Table of contents
Contents Page
Acknowledgments.......................................................................................................................................... i
Summary .....................................................................................................................................................xiii
Samenvatting............................................................................................................................................... xxi
List of abbreviations ................................................................................................................................. xxix
Chapter One: Framework, research objectives and outline of the dissertation ............................................. 1
1.1. Framework of the research ............................................................................................................ 3
1.2. Research objectives ....................................................................................................................... 5
1.3. Hypotheses of the PhD study ........................................................................................................ 6
1.4. Outline of the dissertation ............................................................................................................. 7
Chapter Two: Introduction and general background .................................................................................... 9
2.1. Pesticide use in developing countries ......................................................................................... 11
2.2. Pesticide use and impacts in Ethiopia ......................................................................................... 12
2.3. Pesticide residues in food............................................................................................................ 17
2.4. Human exposure and toxicity of pesticides ................................................................................ 19
2.5. Risk assessment .......................................................................................................................... 20
2.5.1. Hazard identification ........................................................................................................... 21
2.5.2. Exposure assessment ........................................................................................................... 21
2.5.3. Food consumption data ....................................................................................................... 22
2.5.4. Dose-response assessment .................................................................................................. 23
2.5.5. Risk characterization ........................................................................................................... 23
2.5.6. Dietary exposure models for pesticide residues in food ..................................................... 24
2.5.6.1. Deterministic dietary exposure model ............................................................................ 24
2.5.6.2. Probabilistic dietary exposure model ............................................................................. 24
Chapter Three: Pesticide residue evaluation in major staple food items in Ethiopia .................................. 27
Abstract ................................................................................................................................................... 29
3.1. Introduction ................................................................................................................................. 30
3.2. Materials and Methods ................................................................................................................ 33
3.2.1. Sampling ............................................................................................................................. 33
3.2.2. Sample preparation ............................................................................................................. 34
3.2.3. Reagents and materials........................................................................................................ 34
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3.2.4. Analytical equipment .......................................................................................................... 35

3.2.5. Analytical method validations: limit of detection (LOD), limit of quantification (LOQ),
repeatability (% RSD), linearity and recovery .................................................................................... 36
3.2.6. QuEChERS extraction method ........................................................................................... 37
3.2.7. Analysis of food samples .................................................................................................... 38
3.2.8. Data analysis ....................................................................................................................... 39
3.3. Results ......................................................................................................................................... 39
3.3.1. Method validation results .................................................................................................... 39
3.3.2. Pesticide residues in tested food samples............................................................................ 41
3.3.3. Comparison of pesticide residue with the maximum residue limit (MRL) set by Codex
Alimentarius........................................................................................................................................ 44
3.3.4. Pesticide concentration (µg/kg) in lentil (Lens culinaris) and wheat (Triticum spp.) flour 44
3.4. Discussion ................................................................................................................................... 45
3.5. Conclusions ................................................................................................................................. 50
Chapter Four: Risk of DDT to infants consuming maize as complementary diet in southwestern Ethiopia
.................................................................................................................................................................... 51
Abstract ................................................................................................................................................... 53
4.1. Introduction ................................................................................................................................. 54
4.2.1. Study area............................................................................................................................ 55
4.2.2. Dietary survey ..................................................................................................................... 56
4.2.3. Maize sampling ................................................................................................................... 57
4.2.4. Reagents and materials, analytical equipments, extraction and clean-up of the maize
samples, method validation procedures .............................................................................................. 57
4.2.5. Statistical and exposure analysis ......................................................................................... 57
4.2.6. Deterministic exposure assessment ..................................................................................... 58
4.2.7. Probabilistic exposure assessment ...................................................................................... 58
4.3. Results ......................................................................................................................................... 59
4.3.1. Socio demographic results .................................................................................................. 59
4.3.2. Concentration of total DDT and its metabolites in household and market samples ........... 59
4.3.3. Exposure assessment ........................................................................................................... 62
4.3.3.1. Deterministic exposure assessment ................................................................................. 62
4.3.3.2. Probabilistic exposure assessment.................................................................................. 63
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4.3.4. Comparison of EDI of DDT for the total population in the deterministic and probabilistic
exposure assessment ........................................................................................................................... 64
4.4. Discussion ................................................................................................................................... 65
4.5. Conclusions ................................................................................................................................. 67
Chapter Five: Effect of household processing on pesticide residues in coffee (Coffea arabica) and teff
(Eragrostis tef) ............................................................................................................................................ 69
Abstract ................................................................................................................................................... 71
5.1. Introduction ................................................................................................................................. 72
5.2.1. Chemicals and reagents ....................................................................................................... 75
5.2.2. Sampling ............................................................................................................................. 76
5.2.3. Quality control .................................................................................................................... 76
5.2.4. Treatment of raw coffee beans ............................................................................................ 76
5.2.4.1. Raw coffee beans............................................................................................................. 77
5.2.4.2. Washed coffee beans ....................................................................................................... 77
5.2.4.3. Roasted coffee beans ....................................................................................................... 77
5.2.4.4. Brewed coffee .................................................................................................................. 77
5.2.4.5. Raw teff flour................................................................................................................... 78
5.2.4.6. Dough preparation.......................................................................................................... 78
5.2.4.7. Baking ............................................................................................................................. 78
5.2.5. Determination of processing factor (PF) ............................................................................. 78
5.2.6. Analysis of wash water and coffee sludge .......................................................................... 79
5.2.7. Analytical equipment .......................................................................................................... 79
5.2.8. Statistical analysis ............................................................................................................... 80
5.3. Results ......................................................................................................................................... 80
5.3.1. Effect of household coffee processing on pesticide residues .............................................. 80
5.3.2. Determination of the processing factor (PF) ....................................................................... 81
5.3.3. Pesticide residue in wash water and coffee sludge ............................................................. 82
5.3.4. Effect of household processing on pesticide residues in teff .............................................. 83
5.4. Discussion ................................................................................................................................... 85
5.5. Conclusions ................................................................................................................................. 88
Chapter Six: Pesticide residues in drinking water and associated risks to consumers in Ethiopia ............. 89
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Abstract ....................................................................................................................................................... 91
6.1 Introduction ................................................................................................................................. 92
6.2 Materials and Methods ................................................................................................................ 95
6.2.1. Study area............................................................................................................................ 95
6.2.2. Sampling ............................................................................................................................. 97
6.2.3. Chemicals and reagents ....................................................................................................... 98
6.2.4. Extraction and clean-up of the samples .............................................................................. 98
6.2.4.1. Liquid- solid extraction procedure.................................................................................. 98
6.2.4.2. Liquid-liquid extraction procedure ................................................................................. 99
6.2.5. Analytical equipment and chromatographic separation .................................................... 100
6.2.6. Screening........................................................................................................................... 100
6.2.7. Quality control .................................................................................................................. 101
6.2.8. Statistical analysis ............................................................................................................. 101
6.2.9. Risk assessment................................................................................................................. 102
6.2.9.1. Chronic risk assessment ................................................................................................ 102
6.2.9.2. Acute risk assessment .................................................................................................... 103
6.3 Results ....................................................................................................................................... 103
6.3.1 Method validation results ...................................................................................................... 103
6.3.2 Pesticide residue in drinking water sources in Jimma zone, southwest Ethiopia.................. 104
6.3.3 Exposure assessment of population in Jimma zone to pesticide residues in water ............... 106
6.3.4 Pesticide residues in Addis Ababa water sources ................................................................. 107
6.3.5 Exposure assessment of population in Addis Ababa to pesticide residues in water ............. 107
6.4 Discussion ................................................................................................................................. 108
6.5 Conclusions ............................................................................................................................... 111
Chapter seven: Exposure to DDT from khat (Catha edulis) consumption: a double burden to consumers in
southwestern Ethiopia ............................................................................................................................... 113
Abstract ..................................................................................................................................................... 115
7.1 Introduction ............................................................................................................................... 116
7.2 Materials and Methods .............................................................................................................. 118
7.2.1 Study area.............................................................................................................................. 118
7.2.2 Sample size determination for consumption survey ............................................................. 119
7.2.3 Consumption survey ............................................................................................................. 119
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7.2.4 Data handling and analysis ................................................................................................... 120

7.2.5 Khat sampling ....................................................................................................................... 120
7.2.6 Reagents and materials.......................................................................................................... 120
7.2.7 Quality control ...................................................................................................................... 121
7.2.8 Extraction and clean-up of khat samples .............................................................................. 121
7.2.9 Analytical equipment ............................................................................................................ 122
7.2.10 Exposure assessment of khat consumers............................................................................... 122
7.2.11 Probabilistic exposure analysis ............................................................................................. 123
7.3 Results ....................................................................................................................................... 123
7.3.1 Socio demographic results of the khat consumers ................................................................ 123
7.3.2 Occurrence of DDT and its metabolites in khat .................................................................... 124
7.3.3 Exposure assessment ............................................................................................................. 126
7.3.4 Probabilistic exposure analysis ............................................................................................. 127
7.4 Discussion ................................................................................................................................. 127
7.5 Conclusions ............................................................................................................................... 129
Chapter Eight: General discussion, conclusions and future research prospects ....................................... 131
8.1. General discussion .................................................................................................................... 133
8.1.1. Pesticide residues problems in foods .................................................................................... 133
8.1.2. Human exposure and associated risk to consumers .............................................................. 135
8.1.3. Cumulative risk assessment for multiple pesticides detected in commonly consumed foods
137
8.1.4. Importance of food processing for the reduction of pesticide residues in food .................... 143
8.2. Conclusions ............................................................................................................................... 144
8.2. Future research prospects .......................................................................................................... 148
References ................................................................................................................................................. 151
Annexes .................................................................................................................................................... 175
Annex 1: Model out puts from probabilistic exposure analysis of DDT in khat .................................. 177
Annex 2: @risk detailed statistics for the intake of DDT due to consumption of khat from Monte-Carlo
simulation model................................................................................................................................... 179
Annex 3: Curriculum Vitae ................................................................................................................... 181
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Summary
Summary
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Summary
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Summary
Pesticides are indispensible to increase agricultural production and control human and animal
diseases. Due to their benefits, pesticides have been continued to be used worldwide. Even
though, the use of pesticides is important, their residues eventually end up in different
environmental compartments (water, food, soil, air) and may negatively affect human health and
the environment. Because of the harmful effects to human and other organisms in the
environment, pesticides are under strict regulations starting from their synthesis and formulation,
over the mixing/loading of pesticides and their application in the field.
Ethiopia’s population is continuously increasing and this put pressure on agricultural activities.
Due to this pressure, the government of Ethiopia use agricultural intensification with high
demands for fertilizers and pesticides as one of the strategies to increase agricultural production.
Additionally, unlike most developed countries, the pesticide regulation such as pesticide
registration and control proclamation 674/2010 in Ethiopia, is not well enforced due to different
reasons explained in chapter one and chapter two. Moreover, most of the Ethiopian farmers are
illiterate and they do not know how much amount they should apply in the field, where to store
the pesticides and from whom they should buy these chemical pesticides. As a result, the
different consumer products may become contaminated by pesticides. Despite all these
problems, there is no study done on the contamination of the commonly consumed food items
(staple foods) with pesticides in the country. Additionally, there is no data in Ethiopia regarding
human exposure and consumer risk assessment from pesticide exposure. Therefore, it is
important to evaluate the presence of pesticide residues in commonly consumed food items and
assess their risks to consumers.
Chapter three presents, the analysis of the different pesticide residues in food items in
southwest Ethiopia. For this study, teff (Eragrostis tef), maize (Zea mays), red pepper (Capsicum
annuum), coffee (Coffea arabica), were collected from one big central market in Jimma zone,
southwest Ethiopia. This market is chosen as a sample point, because most agricultural crops are
collected for sale. The selection of the pesticides for analysis in food items was done based on
the information obtained from agricultural offices and farmers in the study area. The study also
includes pesticides such as DDT and endosulfan which are known to be applied for indoor
residual spraying (IRS) to control malaria, as Jimma zone is a malaria endemic area. For the
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Summary
analytical procedure, the Quick, Easy, Cheap, Effective, Rugged and Safe (QuEChERS)
extraction method combined with dispersive Solid Phase Extraction (d-SPE) clean up was
applied. Pesticides determination was done using gas chromatography with electron capture
detector (GC-ECD). In this work, different pesticides such as, DDT and its metabolites,
endosulfan (α and β isomers), cypermethrin, permethrin, deltamethrin and chlorpyrifos ethyl
were detected at a concentration range of 0.011 to 1.115 mg/kg food. The findings of this study
indicate that the commonly consumed food items were contaminated by pesticides, which may
come from field application as well as from application for disease control. More than one third
of the pesticides detected were above the maximum residue limit (MRL) set by Codex
Alimentarius. This indicates that there is overuse of the pesticides in the study area. DDT and
endosulfan which are not authorized for agricultural use were also detected in the food items.
This may be due to, cross contamination during spraying for malaria control or may be due to
their persistent (non-degradable) nature in the environment. The detection of these pesticide
residues in the commonly consumed foods may affect human health and food safety in general.
Chapter four presents, the risk of DDT residues in maize to infants in the southwest Ethiopia.
In this region, maize is the commonly produced agricultural crop which is also used as a major
component in infants’ food. To work out a risk assessment of this pesticide to infants, a
consumption and residue data base is needed. Therefore, a consumption survey was undertaken
for 6-12 month infants, from randomly selected households, using the 24 hour recall method.
The maize samples were collected from the households selected for the consumption survey, as
well as from markets in the surrounding area. During screening of the maize samples, DDT and
its metabolites were detected. To assess the exposure of infants, the estimated daily intake (EDI)
was determined using the deterministic and probabilistic exposure analysis method based on the
recommendation of the European Food Safety Authority (EFSA). The EDI was compared with
the health based reference values such as provisional tolerable daily intake (PTDI) to
characterize the risk. From the results, it was found that about three fourth of the maize samples
were contaminated with DDT residues above the MRL norm set by Codex Alimentarius. This
may be due to illegal application of this pesticide in agriculture or due to previous application for
malaria control which is discussed in chapter three. The results of the exposure estimation
revealed that the mean and the high consumer (97.5 percentile) EDIs of the total DDT for the
infants were above the PTDI. This is an indication of chronic health problems for the infants
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consuming maize as a complementary food. The health problem may be severe for this
vulnerable group of the population. This may be due to their metabolic mechanism to detoxify
the chemical hazards is not well matured at that age. In addition to this, the infants’ food intake
per body weight is higher compared to adults. Therefore, continuous monitoring of DDT and
other pesticides in infants food is recommended in Ethiopia.
Several studies report that food processing such as washing, peeling, baking, cooking, roasting,
boiling, fermentation, drying etc. have an effect on the reduction of pesticide residues in different
food commodities. Our study in chapter five presents, the effect of household processing such as
washing, roasting, brewing, doughing and baking on the level of pesticides in coffee and teff.
The previously detected pesticides in teff and coffee (chapter three), such as DDT,
deltamethrin, cypermethrin, permethrin, endosulfan (α and β) and chlorpyrifos ethyl were spiked
in blank samples of teff and coffee beans. Following this, the household processing methods
were undertaken and the effect on the pesticide residues was determined by calculating the
processing factor (PF). According to the joint meeting on pesticide residues (JMPR) of
FAO/WHO, PF is defined as the ratio of the residues in the processed food commodity to the raw
commodity before processing. The result of this study indicates that the PF for each household
preparation of coffee and teff is less than one. This means that there is a reduction of pesticide
residues due to the household processing of these two food commodities. From the processing
methods studied, roasting (reduction of 99.8%) and brewing (reduction of 100%) of coffee and
baking (reduction of 90.2%) of teff were the most effective methods to reduce pesticide residues
of the raw food items. Therefore, in countries like Ethiopia, who do not have well enforced
pesticide regulations and proper pesticide monitoring programs, food processing may be used as
a solution to minimize exposure and the risks associated with pesticide residues in food.
Human exposure to chemical hazards such as pesticides may not happen only from consumption
of food but also from drinking water. Most of the water sources are located in the vicinity of
agricultural fields where pesticides are applied. The water sources is often contaminated due to
runoff, environmental drift, erosion and diffusion processes. This may affect human health and
the environment as well. Chapter six presents the evaluation of different pesticides in water
sources around Jimma zone, southwest Ethiopia and Addis Ababa which is the capital of
Ethiopia. Water samples were collected from springs, wells, river, pond, the distribution
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reservoir and community taps. For the extraction of water samples liquid-solid and liquid-liquid
analytical methods were applied. The results of this study reveal that the water sources were
contaminated with organophosphate pesticides (diazinon, malathion and pirimiphos methyl), a
herbicide (2,4-D) and a fungicide (fenpropimorph) at varying concentration (from 0.109-138
µg/l) from source (river, spring, and well ) up to the point of consumption (community taps). The
concentration of the detected pesticides in the water was higher in the untreated water sources
such as the well, the spring, and the river. This may indicate that the conventional treatment
process such as coagulation/flocculation, filtration and chlorination may have some effect on the
reduction of pesticide level in the water. From our findings, the estimated daily intake (EDI) of
the detected pesticides from drinking water was less than the acute reference dose (EDI < ARfD)
for all the pesticides detected in water. Although, the concentration for diazion and
fenpropimorph was above the acceptable daily intake (ADI). This may result in chronic health
problems for consumers. This suggests also a need for continuous monitoring of pesticides in the
water bodies in Ethiopia.
In addition to the contamination of the above mentioned conventional edible food commodities
and drinking water, pesticides may also detected in other agricultural crops such as khat (Catha
edulis). Khat is consumed in many parts of East Africa (Ethiopia, Somalia, Djibouti, Eritrea and
Kenya) and Arabian countries (Saudi Arabia, Yemen, and Tunisia). Within the communities in
these different countries, khat chewing is considered a social habit. In Ethiopia, about half of the
population consumes the khat plant. However, chewing khat causes different health problems
such as euphoric effect, excitement, constipation, mydriasis (dilated pupil of the eye),
hyperactivity, and suppressed apatite. If khat also contains pesticides it causes a double burden to
consumers, as pesticides have also their own health effects. Chapter seven presents the human
exposure to the commonly detected and highly persistent pesticide DDT from the consumption
of khat in southwest Ethiopia. The chewable part of the khat plant (leaves with tender stems) was
collected from the central market in Jimma zone, southwest Ethiopia. The khat samples were
transported to the laboratory and dried under shade to prevent photodegradation. Solid phase
extraction with column chromathography clean up procedures was undertaken to extract the khat
samples. DDT and its metabolites were detected in 80% of the khat samples. To assess the
human health risk of DDT exposure in khat, a probabilistic exposure analysis method was
applied and compared with the health based reference value (PTDI). The results indicate that the
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estimated daily intake (EDI) of DDT was lower than PTDI. However, this does not guarantee
100% safety as this plant is consumed without any preceding processing steps. Therefore,
creating awareness for consumer population about the double health burden of chewing khat and
continous monitoring of DDT and other pesticide in this plant is important.
In conclusion, the PhD research provides evidence on the level of contamination of the
commonly consumed food commodities, drinking water, and khat by pesticides and associated
risk to human health. Mutual consumption of these commonly consumed food items may expose
the Ethiopian population to multiple pesticides which results in a cumulative risk. Chapter eight
presents, the general discussion and implication of the findings for human exposure and
consumer risk, conclusions and recommendations for further research.
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Samenvatting
Samenvatting
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Samenvatting
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Samenvatting
Bestrijdingsmiddelen zijn onmisbaar om de landbouwproductie te verhogen en voor het

beheersen van ziektes bij mens en dier. Omwille van hun voordelen worden bestrijdingsmiddelen
wereldwijd gebruikt. Hoewel het gebruik van bestrijdingsmiddelen belangrijk is, blijven hun
residuen in verschillende milieucompartimenten aanwezig (water, bodem, lucht) en kunnen zij
de menselijke gezondheid en het milieu negatief beïnvloeden. Omwille van hun schadelijke
effecten voor mens en andere organismen, staan bestrijdingsmiddelen onder strikte regulering
vanaf hun synthese en formulering over het aanmaken en laden van het spuitmengsel en hun
toepassing op het veld.
Door de steeds groeiende populatie in Ethiopië staat de landbouw onder druk. Daardoor maakt
de regering van Ethiopië gebruik van intensifiëring van de landbouw, wat gepaard gaat met een
hoge vraag naar meststoffen en bestrijdingsmiddelen als een van de strategieën om de
landbouwopbrengst te verhogen. Bovendien wordt, in tegenstelling tot de meeste ontwikkelde
landen, de wetgeving i.v.m. bestrijdingsmiddelen, waaronder registratie en controle
(Proclamation 674/2010), in Ethiopië niet zo goed afgedwongen door verschillende redenen die
beschreven worden in hoofdstuk 1 en hoofdstuk 2. Daarenboven zijn de meeste telers in
Ethiopië analfabeet en weten dus niet goed hoeveel bestrijdingsmiddelen ze dienen toe te passen
op het veld, waar ze deze middelen dienen te stockeren en waar de middelen aan te kopen.
Bijgevolg kunnen verschillende consumentenproducten gecontamineerd geraken met schadelijke
bestrijdingsmiddelen. Niettegenstaande deze problemen, is er in het land nog geen studie
uitgevoerd waarbij de contaminatie van vaak geconsumeerde voedingsmiddelen met
bestrijdingsmiddelen werd onderzocht. Bovendien is er in Ethiopië geen data beschikbaar i.v.m.
humane blootstelling en consumenten risico analyse van blootstelling aan bestrijdingsmiddelen.
Daarom is het belangrijk om de aanwezigheid van residu’s van bestrijdingsmiddelen in vaak
geconsumeerde voedingsmiddelen te evalueren en hun risico’s voor de consument in te schatten.
Hoofdstuk 3 handelt over de analyse van verschillende bestrijdingsmiddelenresidu’s in het

zuidwesten van Ethiopië. In deze studie werden teff (Eragrostis tef), maïs (Zea mays), rode peper
(Capsicum annuum) en koffie (Coffea arabica) verzameld in een grote centrale markt in de zone
Jimma, in het zuidwesten van Ethiopië. Deze markt werd gekozen als locatie voor staalnames
omdat de meeste gewassen er te koop worden aangeboden. De selectie van bestrijdingsmiddelen
voor analyse in voedingsmiddelen gebeurde op basis van informatie bekomen via
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Samenvatting
landbouwkantoren en telers in het studiegebied. De studie omvat ook bestrijdingsmiddelen als

DDT en endosulfan, welke gekend zijn omwille van hun gebruik binnenshuis (IRS) om malaria
te bestrijden, Jimma is immers een gebied waar malaria heerst. De analytische procedure werd
uitgevoerd met behulp van een ‘Quick, Easy, Cheap, Effective, Rugged and Safe’ (QuEChERS)
extractie methode gecombineerd met vastefase extractie (d-SPE) opzuivering De bepaling van
bestrijdingsmiddelen werd uitgevoerd met behulp van gas chromatografie met elektronendetector
(GC-ECD). In dit werk werden verschillende bestrijdingsmiddelen, zoals DDT en zijn
metabolieten, endosulfan (α en β isomeren), cypermethrin, permethrin, deltamethrin en
chloorpyrifos-ethyl gedetecteerd aan een concentratie-interval van 0,011 tot 1,115 mg/kg
voedsel. De bevindingen uit onze studie geven aan dat de courant geconsumeerde
voedingsmiddelen gecontamineerd waren met bestrijdingsmiddelen, afkomstig uit zowel
toepassing in het veld als uit toepassing ter bestrijding van ziekten. Meer dan een derde van de
gedetecteerde bestrijdingsmiddelen overschreden de maximum residu limiet (MRL), bepaald
door de Codex Alimentarius. Dit geeft aan dat er te veel bestrijdingsmiddelen worden gebruikt in
het bestudeerde gebied. DDT en endosulfan, welke niet toegelaten worden voor landbouwkundig
gebruik, werden ook teruggevonden in voedingsmiddelen. Dit kan te wijten zijn aan kruis-
contaminatie tijdens het spuiten tegen malaria of aan hun persistent karakter in het milieu.
Detectie van deze residuen in vaak geconsumeerde voedingsmiddelen kunnen de menselijke
gezondheid en de voedselveiligheid in het algemeen, nadelig beïnvloeden
Hoofdstuk 4 beschrijft de risico’s van DDT residuen in maïs voor jonge kinderen in het
zuidwesten van Ethiopië. Maïs is in dit gebied een veelvuldig geproduceerd gewas dat ook een
belangrijk bestanddeel is in de voeding van jonge kinderen. Om een risicoanalyse van dit middel
uit te werken voor kinderen zijn data over consumptie en residuen nodig. Een consumptie
enquête werd dus uitgevoerd bij jonge kinderen tussen 6-12 maanden oud uit willekeurig
geselecteerde gezinnen, gebruik makend van de methode waarbij men bijhoudt wat men de
afgelopen 24 u heeft geconsumeerd. De maïsstalen werden verzameld vanuit de huishoudens en
vanuit lokale markten in het omliggende gebied. Bij screening van de maïsstalen werden DDT en
zijn metabolieten gedetecteerd. Om de blootstelling van jonge kinderen in te schatten werd de
geschatte dagelijkse inname (EDI) bepaald, gebruik makende van de deterministische en
probabilistische blootstellingsanalyse methode gebaseerd op aanbevelingen van het EFSA
(European Food Safety Authority). De EDI werd vergeleken met referentiewaarden zoals de
xxiv
Samenvatting
toelaatbare dagelijkse inname norm (PTDI) om het risico te bepalen. De resultaten gaven aan dat
ongeveer ¾ van de maïsstalen gecontamineerd waren met residuen van DDT boven de MRL
norm, zoals bepaald door de Codex Alimentarius. Dit kan het gevolg zijn van illegale toepassing
in de landbouw of van eerdere toepassingen tegen malaria (hoofdstuk 3). De resultaten van de
blootstellingsschatting toonden aan dat de gemiddelde en hoge consumenten (97,5 percentiel)
EDI’s van het totale DDT voor de kinderen boven de PTDI zaten. Dit is een indicatie voor
chronische gezondheidsproblemen voor de jonge kinderen die maïs eten als aanvullende
voeding. Het risico kan ernstig zijn voor deze kwetsbare populatiegroep, mogelijks doordat hun
fysiologie op die leeftijd nog niet in staat is om deze chemische gevaren te metaboliseren en
efficiënt uit te scheiden. Bovendien is hun voedselinname per lichaamsgewicht hoger dan bij
volwassenen. Bijgevolg is de continue monitoring van DDT en andere bestrijdingsmiddelen in
Ethiopië aangewezen.
Verschillende studies rapporteren dat de huishoudelijke verwerking van levensmiddelen, zoals

wassen, schillen, bakken, stoven, roosteren, koken, fermenteren, drogen, enz. een effect hebben
op de vermindering van residuen van bestrijdingsmiddelen in voedsel. Onze studie in hoofdstuk
5 geeft het effect aan van verwerkingsmethoden als wassen, roosteren, brouwen, maken van deeg
en bakken op de hoeveelheid bestrijdingsmiddelen in koffie en teff. De voordien gedetecteerde
bestrijdingsmiddelen in koffie en teff (hoofdstuk 3), waaronder DDT, deltamethrin,
cypermethrin, permethrin, endosulfan (α en β) en chloorpyrifos-ethyl werden in gekende
hoeveelheden toegevoegd aan teff en koffiebonen. Hierop volgend werden de huishoudelijke
verwerkingsmethoden uitgevoerd en de residuen bepaald door de verwerkingsfactor (PF) te
berekenen. Volgens de joint meeting over bestrijdingsmiddelenresiduen (JMPR) van het
FAO/WHO wordt de PF gedefinieerd als de verhouding van residuen in verwerkt voedsel tot de
rauwe voedingsgrondstof voor de verwerking. Het resultaat van de studie geeft aan dat voor
zowel de verwerking van koffie en teff de verwerkingsfactor (PF) lager is dan één, wat een
indicatie is voor een reductie van bestrijdingsmiddelenresiduen vanwege de toegepaste
voedselverwerking. Van de bestudeerde huishoudelijke bewerkingsmethoden waren voor
koffiebonen het roosteren (99,8% residudaling) en het koffiezetten (100% residudaling) en voor
teff het bakken (residudaling van 90.2%) het meest effectief om de residuen van
bestrijdingsmiddelen te doen afnemen in de rauwe voedingsmiddelen. Daarom kan in landen als
Ethiopië, die niet beschikken over een sterke en gehandhaafde bestrijdingsmiddelen regelgeving
xxv
Samenvatting
en monitoringprogramma's, de verwerking van levensmiddelen één van de oplossingen zijn om

de risico's in verband met bestrijdingsmiddelen te beperken.
Humane blootstelling aan chemische gevaren waaronder bestrijdingsmiddelen kan niet alleen
gebeuren via voedsel maar ook via drinkwater. De meeste waterreservoirs liggen in de nabijheid
van landbouwgronden, waar bestrijdingsmiddelen worden toegepast. Het oppervlaktewater is
veelal gecontamineerd via uitspoeling, milieudrift, erosie of diffusieprocessen. Dit kan zowel het
milieu als de volksgezondheid schaden. Hoofdstuk 6 evalueert verschillende
bestrijdingsmiddelen in waterbronnen uit de Jimma zone, in zuidwest Ethiopië, en Addis Ababa,
de hoofdstad van Ethiopië. Waterstalen werden gecollecteerd uit bronnen, putten, een
waterzuiveringsinstallatie (voor en na zuivering), en verdeelreservoirs en aftakpunten voor
drinkwater, gebruik makende van de “grab sampling” methode. Voor de extractie van de
waterstalen werd gebruik gemaakt van vaste stof-vloeistof en vloeistof-vloeistof analytische
methoden. De resultaten van de studie tonen aan dat de waterstalen gecontamineerd waren met
organofosfaten (diazinon, malathion en pirimifos-methyl), een herbicide (2,4-D) en een
fungicide (fenpropimorph) in variabele concentraties (0,109 - 138 µg/l) vanuit oorsprong (rivier,
bron en put) tot aan het aftakpunt voor drinkwater. De concentratie van gedetecteerde
bestrijdingsmiddelen in het water was hoger in de ongezuiverde waterbronnen zoals de put, bron
en rivier. Dit kan aangeven dat conventionele zuiveringsprocessen zoals coagulatie/flocculatie,
filtratie en chlorinatie een zeker effect hebben op de reductie van de concentratie
bestrijdingsmiddelen in het water. Uit onze bevindingen bleek de geschatte dagelijkse inname
(EDI) van deze bestrijdingsmiddelen uit drinkwater lager dan de acute referentie dosis
(EDI<ARfD) voor alle bestrijdingsmiddelen gedetecteerd in het drinkwater. Echter de
concentratie diazinon en fenpropimorf waren hoger dan de toelaatbare dagelijkse inname (ADI).
Dit kan leiden tot chronische gezondheidsproblemen voor consumenten. Dit suggereert tevens
een nood aan frequente monitoring van verschillende waterbronnen in Ethiopië op aanwezigheid
van bestrijdingsmiddelen.
Naast contaminatie van bovenvermelde conventioneel eetbare voedingsmiddelen en drinkwater,

kunnen bestrijdingsmiddelen ook gedetecteerd worden in andere landbouwgewassen zoals khat
(Catha edulis). Khat wordt in vele gebieden van Oost-Afrika (Ethiopië, Somalië, Djibouti en
Kenia) en Arabische landen (Saudi-Arabië, Jemen en Tunesië) geconsumeerd. Binnen de
xxvi
Samenvatting
gemeenschappen in deze landen wordt het kauwen van khat beschouwd als een sociale
gewoonte. Deze stimulerende plant wordt door ongeveer de helft van de Ethiopische bevolking
geconsumeerd. Echter, het kauwen van khat veroorzaakt verschillende gezondheidsproblemen
waaronder euforische effecten, opwinding, constipatie, mydriasis (verwijding van de oogpupil),
hyperactiviteit en verminderde trek. Als khat ook bestrijdingsmiddelen bevat, heeft dit een
dubbele impact op de consument daar bestrijdingsmiddelen ook hun eigen effect hebben op de
gezondheid. Hoofdstuk 7 gaat over de humane blootstelling aan het courant gedetecteerde en
zeer persistente bestrijdingsmiddel DDT ten gevolge van de consumptie van khat in het
zuidwesten van Ethiopië. De eetbare delen van khat (bladeren met zachte steel) werden
verzameld in de centrale markt in de Jimma zone, in het zuidwesten van Ethiopië. De khat stalen
werden naar het laboratorium gebracht en werden gedroogd in de schaduw om fotodegradatie
tegen te gaan. Vastefase-extractie opzuiveringsprocedures werden toegepast om de khat stalen te
extraheren. DDT en diens metabolieten werden in 80% van de khat stalen gedetecteerd. Om het
gezondheidsrisico van DDT blootstelling in khat in te schatten, werd een probabilistische
blootstellingsanalysemethode toegepast en vergeleken met de gezondheidsgebaseerde referentie
waarde (PTDI). De resultaten gaven aan dat de geschatte dagelijkse inname (EDI) van DDT
lager was dan PTDI, hoewel dat geen 100% veiligheid garandeert, gezien de plant vers wordt
geconsumeerd. Daarom is het belangrijk om de consumenten bewust te maken van de dubbele
gezondheidsimpact die het kauwen van khat met zich meebrengt en om DDT en andere
bestrijdingsmiddelen in deze plant te monitoren.
Om te besluiten, het doctoraatsonderzoek voorziet bewijs van de hoeveelheid contaminatie van

courant geconsumeerde voedingsmiddelen, drinkwater en khat door bestrijdingsmiddelen en de
daarmee geassocieerde risico’s voor de volksgezondheid. Consumptie van deze courante
voedingsmiddelen kan de Ethiopische bevolking blootstellen aan verschillende
bestrijdingsmiddelen, wat resulteert in een gecumuleerd risico. Hoofdstuk 8 behandelt de
globale discussie en implicatie van de resultaten i.v.m. humane blootstelling en
consumentrisico’s, conclusies en aanbevelingen voor verder onderzoek.
xxvii
Samenvatting
xxviii
List of abbreviations
xxix
xxx
ACN Acetonitrile
ADI Acceptable Daily Intake
ARfD Acute Reference Dose
BW Body Weight
CRA Cummulative Risk Assessment
CAG Cumulative Assessment Group
CMG Common Mechanism Group
DDD Dichloro-Diphenyl –Dichloro -Ethane
DDE Dichloro-Diphenyldichloro-Ethylene
DDT Dichloro-Diphenyl- Trichloroethane
EC European Commission
ECD Electron Capture Detector
EDI Estimated Daily Intake
EFSA European Food Safety Authority
EPA Environmental Protection Agency
EEPA Ethiopian Environmental Protection Authority
ERA Environmental Risk Assessment
ESI Electron Spray Ionization
FAO Food and Agricultural Organization
FFQ Food Frequency Questionnaire
GAP Good Agricultural Practice
GC Gas Chromatography
GPS Geographical Positioning System
HCH HexaChloroHexane
HPLC High Performance Liquid Chromatography
xxxi
IRS Indoor Residual Spray
ISARC International Sociological Association Research Committee
KOW Octanol-Water Partition Coefficient
LC Liquid Chromatography
LOD Limit of Detection
LOQ Limit of Quantification
LOAEL Lowest Observed Adverse Effect Level
MCRA Monte-Carlo Risk Assessment
MRL Maximum Residue Limit
MS Mass Spectrometer
ND Non-Detected
OCPs Organochlorine Pesticides
PF Processing Factor
POPs Persistent Organic Pollutants
POD Point of Departure
PPEs Personal Protective Equipments
PSA Primary Secondary Amine
PTDI Provisional Tolerable Daily Intake
QuECHERS Quick, Easy, Cheap, Effective, Rugged and Safe
RA Risk Assessment
RSD Relative Standard Deviation
RPF Relative Potency Factor
RV Reference Value
SPE Solid Phase Extraction
WHO World Health Organization
xxxii
Chapter One: Framework, research objectives and outline of the dissertation
1
2
1.1. Framework of the research
Food safety regarding pesticide residues is an important issue of concern considering consumers
health (PAN UK, 2002). In developing countries, the main concern of the government is to
increase agricultural production by the application of external inputs such as fertilizers and
pesticides (Rundgren, 2006). Access to safe and sufficient food is a basic requirement for human
health. However, ensuring food safety and security becomes a worldwide problem for
governments, commercial organizations and individuals (Brijnath et al., 2014; Kaferstein, 1997).
Pesticides have a great positive impact to control pests and to increase agricultural production;
but they are also identified as a treat to food safety due to their potential harmful effects to
human health and the environment. Some of the general harmful health effects are, damage to
immune system, endocrine disruption, neurodevelopment delays, cancer and respiratory distress
(Andersson et al., 2014).
In most developing countries, human exposure to pesticides may be aggravated due to different
reasons. One of the main reasons is the legislation and regulations with regard to pesticide use
which is not well enforced in developing countries. In Ethiopia there is a regulation regarding
pesticide importation, handling and use which is called the ‘pesticides registration and control
proclamation no.674/2010’ (Yohannes, 2010). However, there is an inadequate implementation,
weak monitoring and follow-up activities, a lack of integration, weak institutional setup, and
poor networking and exchange of information among stakeholders to enforce the regulation.
Additionally, there is illegal use of non-authorized pesticides such as DDT which were banned
for agricultural use. The farmers do not have enough training about how to use pesticide, where
to store, how much amount they have to apply in the field and the precautions before application
(Mekonnen and Agonafir, 2002). The farmers buy cheap peasticides such as DDT from illegal
retailers because they can get it from neighbouring countries.
Moreover, as Ethiopia is one of the tropical countries, insects are common pests that can affect
agricultural crops. According to Aktar et al. (2009), the commonly applied pesticides in tropical
countries are insecticides which are more toxic compared to fungicides and herbicides. Vector
borne diseases such as malaria is also the most prevalent in Ethiopia and insecticides including
DDT were used as indoor residual spray (IRS).
3
Despite all these problems, there is no study that addresses the consumer exposure to pesticides
and associated risks to their health. Therefore, it is the intention of this PhD study to perform a
detailed analysis of pesticide residues in commonly consumed food commodities and drinking
water and undertaking human exposure assessment. For this purpose, the commonly consumed
food items such as teff (Eragrstis tef), red pepper (Capsicum annuum), maize (Zea maize), coffee
(Coffea arabica) and khat (Catha edulis) were collected from the central market in Jimma town,
southwest Ethiopia and water samples were taken from different sources (river, spring, well,
pond and community taps). Following this, a proper analytical method which is environmental
friendly Quick, Easy, Cheap, Effective, Rugged and Safe (QuEChERS) method with dispersive
Solid Phase Extraction (d-SPE) clean-up was applied for the analysis of food samples. Liquid-
solid and liquid-liquid extraction methods were applied for water samples and solid phase
extraction was used for the pesticide extraction from khat samples. The pesticides to be analysed
were selected by interviewing stakeholders (agricultural officers or farmers) about the commonly
applied pesticides in the study area. Some of the pesticides were also included in the study based
on the condition of the study area. As Jimma zone, which is located in the southwestern Ethiopia,
is indeed a malaria endemic area, pesticides such as DDT and endosulfan may be used for vector
control. The determination of pesticides was done using gas chromatography with electron
capture detector (GC-ECD) and liquid chromatography with double mass spectrometer (LC-
MS/MS) analytical instruments.
To understand and make informed decisions to reduce food safety related problems, risk analysis
has become the main focus in food science in recent years. Risk analysis involves three
components: risk assessment (scientific advice and analysis of information), risk management
(regulation and control) and risk communication.
In this study, deterministic and probabilistic exposure assessment models were used to determine
the exposure and risks of pesticides to human health, based on the recommendation of the
European Food Safety Authority (EFSA, 2012a). The data required for these models are the
residues of pesticides in food and the amount of food consumed. The consumption data were
collected using the 24 hour recall method for two non-consecutive days.
After determining the presence of pesticide residues in food and their possible health risks for
consumers, this study also considers whether household food processing such as washing,
4
roasting, brewing, doughing, and baking has an effect on the pesticide residues in food. Two
commonly consumed and exported crops (coffee and teff) were selected and household
processing were undertaken. The mutual consumption of the different food items may cause a
cumulative effect on the health of consumers. Therefore, cumulative risk assessment was also
addressed. Finally, the results were discussed, concluded and a recommendation for future
research was forwarded
1.2. Research objectives
The overall objective of the study is to investigate human exposure and consumer risk
assessment to pesticide use in Ethiopia.
Specifically the study aimed to:
 give the state of the art on pesticide contamination in agricultural crops and water
sources,
 evaluate the presence of different pesticide residues in commonly consumed food
commodities and drinking water in southwest Ethiopia,
 assess the exposure of human beings to pesticides in food, drinking water and khat,
 identify the human health risk by comparing the exposure level with the health based
reference values such as acceptable daily intake (ADI) and acute reference dose (ARfD),
 identify the cumulative risk of multiple pesticides from mutual consumption of food
commodities under study, and
 investigate the effect of household food processing on pesticide residues.
5
1.3. Hypotheses of the PhD study
The following hypotheses have been put forward:
 Pesticide use in Ethiopia increases. High concentration of pesticide residues can be

detected in commonly consumed food commodities such as teff, maize, coffee and red
pepper.
 Infants are the most vulnerable groups of the population. Exposure to pesticides in their
complementary food has a health risk.
 Household processing of coffee and teff such as washing, roasting, brewing, doughing
and baking reduce pesticide residues.
 Drinking water sources such as springs, wells, rivers and tap water may be contaminated
with pesticides. A health risk for consumers is expected.
 Khat consumption is a common practice in Ethiopia. Hence, khat chewers are exposed to
pesticide residues to a level which causes a health risk.
The PhD study intended to answer a couple of research questions (RQ): What is the problem
related to pesticide use in developing countries, particularly in Ethiopia (RQ1)? Does the
commonly consumed food commodities produced in southwest Ethiopia contain pesticide
residues (RQ2)? Are people exposed to different types of pesticides detected in the agricultural
crops (RQ3)? How safe is the level of pesticide exposure from food, water and khat consumption
(RQ4)? Do pesticides cause a health risk for short and long term exposure when residues are
compared to health based reference values (RQ5)? Does the detection of pesticides in drinking
water cause a health problem to consumers (RQ6)? Is there a human exposure from consumption
of conventionally non-edible plant such as khat (RQ7)? Does household processing such as
washing, roasting and brewing of coffee and, doughing and baking of teff has an effect on
residues of pesticide (RQ8)? Is there a cumulative risk for consumers from mutual consumption
of different food items (maize, teff, red pepper, coffee and khat) and drinking water in Ethiopia
(RQ9)? If the use of pesticides causes human risks, what will be the next step (RQ10)?
6
1.4. Outline of the dissertation
Chapter one deals about the framework of the research which explains the concerns of food
safety in relation to pesticides use, objectives of the research including hypotheses of the study,
research questions and dissertation outline. Chapter two describes an overview of pesticide use in
developing countries, pesticide residues in food and exposure assessment to pesticides. Chapter
three deals with a case study about evaluation of pesticide residues in commonly consumed food
items in southwest Ethiopia. Chapter four describes the risk of DDT (the most persistent and
bioaccumulative pesticide in human body) to infants from consumption of maize as a
complementary diet. Chapter five describes the effect of household food processing such as
washing, roasting, brewing of coffee, and doughing and baking of teff on pesticide residues.
Chapter six elaborates an assessment of pesticide residues in drinking water and associated
consumer risks. Chapter seven gives human exposure to DDT residue in khat (Catha edulis) in
Southwest Ethiopia. Lastly, chapter eight integrates the findings from previous chapters,
elaborates possible implications of the findings for human exposure to pesticides in relation to
the findings of other scholars, conclusions and suggestions for future research directions.
Schematic outline of the dissertation is depicted in figure 1.1.
7
Chapter One:
Research objectives and outline of the dissertation
Aim: Identify the goal of the research, concepts of the whole thesis and outline of dissertation
Chapter Two:
Introduction and general background
Aim: Identify the problem regarding pesticide contamination in consumer products
Method: Review of literatures (Web of Science – grey literature)
Chapter Three:
Pesticide residues in commonly consumed foods in Ethiopia
Aim: To evaluate the presence of different pesticides in food
Method: Extraction was done using QuEChERS method with dispersive Solid Phase Extraction
(d-SPE)
Chapter Four: Chapter Five:

Risk of DDT to infants from maize Effect of household processing on pesticide residues
consumption Aim: Evaluate the effect of household processing on
Aim: Investigate the risk of DDT level of pesticide
residue to infants from maize Method: Household processing such as washing,
consumption roasting and brewing for coffee and doughing and
Method: Deterministic and backing for teff were undertaken.
probabilistic method using Monte- Calculating processing factor (PF) to determine the
Carlo simulation model effect of food processing
Pesticide residues in drinking water and associated risks

Chapter Six: Aim: Investigate estimated daily intake of pesticide from drinking water
Method: Liquid- solid and liquid- liquid extraction
Human exposure to DDT from consumption of khat

Chapter Seven: Aim: Determine human exposure to DDT from consumption of khat
Method: Probabilistic exposure analysis using Monte-Carlo simulation model
Chapter Eight:
General discussion, conclusions and recommendations for future research
Aim: Elaborate main findings in relation to other studies, conclude major results and
recommend for future research
Figure 1.1. Schematic outline of the dissertation
8
Chapter Two: Introduction and general background
9
10
2.1. Pesticide use in developing countries
For decades, pesticides protect agricultural crops against damages from pests and diseases in the
field or in storage areas. In addition, pesticides are of paramount importance to control vector
borne human and animal diseases (Handford et al., 2015). As a result, the use of pesticides has
become a common practice in vector control and in modern agriculture to enhance crop yields,
protect the nutritional integrity of food, to facilitate food storage, to assure year-round food
supplies, and to provide attractive food products (Wong et al., 2014).
At the present time, pesticides are more valued in developing countries than before, particularly
the tropical countries who seek to enter the global market by providing valuable crops (Ismail et
al., 2011). However, these goals perhaps may not be easily achieved without the use of
additional input such as pesticides, principally insecticides, herbicides and fungicides, which
were not used in traditional agricultural practices (Ecobichon, 2001). Due to the needs, many of
the older, non-patented, not expensive chemical pesticides such as DDT, is still illegally used by
small scale farmers in most developing countries. Some African countries also move back to
DDT to reduce malaria incidence (Hecht, 2004). DDT is banned as agricultural insecticide
because of its persistent and bio-accumulative nature in the food chain, causing negative human
health and environmental impacts. The Stockholm convention which was held in 2001, included
DDT as one of the 12 persistent organic pollutants to be banned worldwide for agricultural use,
while the use for disease vector control is only possible under strict guidelines (UNEP, 2001).
Moreover, in developing countries, there is a shift from extensive to more intensive land use
(Nesheim et al., 2014). This development was usually associated with the simplification of
ecosystem, irrigation of agricultural lands and use of fertilizers and pesticides. From a study done
in Thailand (Riwthong et al., 2015), the process of agricultural intensification in developing
countries indicates that farmers who cultivate their agricultural lands more intensively use a
larger number and quantity of synthetic pesticides than farmers who do not intensify their
agricultural lands. As a result of intensive use of pesticides some farmers experience adverse
health effects such as dizziness, nausea and vomiting.
Pesticides should be regulated to protect the risks to human being, animals and the environment
(Handford et al., 2015). Even though, pesticides are important in developing countries, the need
11
to ensure local agricultural production and food security while simultaneously protecting the
population against health effects, emerge as a major public challenge. Developing countries use
only 20% of the world’s agrochemicals, while they suffer 99% of the deaths (Donald et al.,
2007). Improper pesticide use, such as haphazard application on agricultural fields, or crop
harvesting without respecting pre-harvest intervals lead to pesticide contamination of the
agricultural produce. Due to these problems, there is human exposure to pesticides through
ingestion of food, drinking water and animal products because of bioaccumulation of specific
pesticides, through breathing of contaminated air as well as to skin contact (Ennaceur et al.,
2007; Issa et al., 2010; Trautmann et al., 2012; Gebremichael et al., 2013; Andersson et al., 2014;
Bakırcı et al., 2014). Most developing countries experience poor application techniques,
inappropriate spraying equipment and improper storage practices (Damalas and
Eleftherohorinos, 2011).
In Africa, the usage of pesticides continuously increases in recent years (Williamson et al.,
2008). In comparison to other parts of the world, a low volume of pesticide use was noticed in
Africa. This does not mean low environmental contamination or low health risks, as some of the
most toxic pesticides are still often applied under extremely dangerous conditions. Furthermore,
the problem of pesticides is aggravated due to less enforcement of regulation as the national
regulatory agencies are underfunded and lack of resources is available and some African
countries also may not properly register or authorize the pesticide products.
Moreover, there is a lack of guidelines for the control and monitoring of pesticides. Insufficient
knowledge about pesticides, poor equipped laboratory facilities, and inadequate storage are some
of the additional problems in Africa (Matthews et al., 2011). In general, developing countries
lack awareness about proper management and associated risks of pesticides, have a lack of
trained man power, a lack of disposal facilities, etc. which result in adverse impacts of pesticides
on human health and environment.
2.2. Pesticide use and impacts in Ethiopia
Ethiopia is a country of huge biodiversity and agricultural complexity. As many developing

countries, agriculture is the foundation of Ethiopian economy which supports the livelihood of
12
the people. Agriculture accounts for 46.3% of the gross domestic product, 60% export and 80%
of employment (FAO, 2011). The agricultural production in Ethiopia focuses mainly on cereals
(84.55%), followed by pulses (11.13%) and other fresh produce (4.32%) such as fruits and
vegetables (FAO, 2006). The cereals are mainly produced for domestic consumption with only
small market surpluses. The five most commonly produced cereal crops account for almost a
quarter of the cultivated land and about 68% of the total production are teff, maize, wheat,
sorghum and barley (Taffesse et al., 2014).
Ethiopia is also known to be the world fifth and Africa’s top producer of coffee. The country
leads Africa’s domestic consumption. Half of the coffee produced is locally consumed
(Amamo, 2014). This reflects the cultural importance of coffee for the population of Ethiopia.
On the other hand, coffee is considered as the main export commodity in the country, which
provides 31% of the foreign exchange. About 25% of the Ethiopian population is directly or
indirectly dependent on coffee for their income.
In Ethiopia, farmers practiced rain feed agriculture and their dependency on rain may have an
impact on the agricultural production and also on food security (Food and agricultural
organization of the united nation, 2014). Ethiopia is the second most populous country in Africa
with a total estimated population of more than 99,465,819 in July 2015 (CIA, 2015). The
national grain production is not enough to feed the whole population (Stokes et al., 2010).
Ethiopian agriculture is highly influenced by geographical locations. The landscape in the
country defines the potential of agricultural production, the access to input and output markets
and the density of the local population, which determine the local demand in food (Chamberlin
and Schmidt, 2011). Due to this, the government of Ethiopia applies agricultural intensification
as one of the strategies which demands the application of fertilizers and chemical pesticides.
Chemical pesticides were introduced in Ethiopia since 1960s and the introduction was in
connection to the development of commercial farms (EPA, 2004). Currently, the Ethiopian
government initiated to make the country food self-sufficient and to increase agricultural export
commodities like coffee, flowers and vegetables, to maximize the diversity by use of agricultural
intensification. Additionally, pressure exercised by pesticide sale supporting groups makes the
farmers to believe that pesticides are the only way to avoid crop losses.
13
According to Abiye and Hadera, (2005), Ethiopia imports more than 3800 tons of pesticides per
year and the country also obtains pesticides by donations. From all the pesticides imported about
72% are insecticides, 25% are herbicides, 2.6% are fungicides and 1.3% are other products such
as rodenticides and disinfectants. The country’s legalization for pesticide registration and a
monitoring system were introduced in the late 1990’s. Although there is legislation governing
pesticide registration, clear guidelines on the importation, testing, and use of pesticides have not
been effectively enforced. Therefore, it is common to find restricted or banned pesticides widely
used in Ethiopia. There is also poor application practice without using personal protective
equipments (PPEs). This results in unprotected spreading of pesticides into the environment and
which causes human exposure to pesticides. Some bad practices of bystander exposure and
unprotected spray operators are shown in figure 2.1.
Figure 2.1. Outdoor (A) and indoor or greenhouse (B) pesticide application in Ethiopia
In Ethiopia, there is a massive use of pesticides on large scale farms (Westbom et al., 2008)
while, the use of pesticides in small scale farming is less. However, the government extension
services promote the use of pesticides to improve productivity and to reduce food insecurity for
the Ethiopian people living from small scale farming (Environment and Social Assessment
International, 2006). Besides the agricultural use, pesticides have been used for public health to
control vector borne diseases such as malaria. Indoor residual spraying (IRS) is used to prevent
malaria in epidemic prone areas (Ministry of Health, 2006). For the last 40 years, Ethiopia has
14
primarily used DDT for its IRS operation program (Biscoe et al., 2004). The country applies
approximately 400 metric tons of the active ingredients DDT per year (Sadasivaiah et al., 2007;
WHO, 2007; van den Berg, 2009). Nowadays, the use of DDT for malaria vector control is
phased out due to the resistance development by the malaria vector and replaced by effective
insecticides such as primiphos-methyl and propoxur (Balkew et al., 2012). Additionally,
organochlorine pesticides (OCPs) such as DDT, which were used widely in agriculture and for
public health purposes, became a worldwide concern due to their persistence, bioaccumulation
potential through food chain, and health effects to human and non-target organisms ( Jones and
de Voogt, 1999; Donaldson et al., 2010). Ethiopia built-up the largest stockpiles of obsolete
pesticides in Africa. These stocks have been accumulated starting from the first imports in the
1960s. The stock mostly contains old and toxic organochlorine compounds such as DDT,
chlordane, dieldrin and lindan. There are different reasons for the accumulation of obsolete
stocks in Ethiopia. The major reason is excessive and uncoordinated donations of pesticides by
donors and purchases by Ethiopian government. For example, out of 93 tons of obsolete
pesticide stocks inventoried in 2005, 67% were obtained by donation, while the rest were
imported by the Ethiopian government (Abiye and Hadera, 2005). Figure 2.2 shows obsolete
pesticide stocks in different African countries in which Ethiopia holds more than 1000 tons.
Figure 2.2. Obsolete pesticide stocks in Africa (source: (FAO, 2005)
15
Small scale peasant farming and some large scale mechanized agriculture in Ethiopia consider
pesticides as a best choice for improving agricultural production and quality of life in general.
However, evidences in the last few decades have shown that the used pesticides can also be
detrimental to human health and the ecosystem (Amera and Abate, 2008). Some of the
underlying reasons for these problems are:
o Lack of awareness: illiteracy of the majority of the peasants, inadequate training, lack of
appropriate and timely information about the proper use and management of pesticides,
inappropriate use of personal protective equipment (PPE), wrong notion that pesticides are
the best solution to pest control, and poor guidance about the safe use and handling;
o Inappropriate use of pesticides: wrong mix of different types of pesticides, use of
pesticides for unintended purposes, and utilization of empty pesticide containers for
domestic purposes;
o Weak law enforcement: late issuance of regulations and guidelines, inadequate
implementation of the issued regulations, weak monitoring or follow-up activities to
control pesticide usage;
o Disposal problems: there is no disposal facility in Africa; particularly in Ethiopia due to
the high cost of disposal and the cost of appropriate destruction of obsolete pesticides is
too high.
Due to these problems, the contamination of the different environmental compartments (water,
air, soil, food and biota) by pesticides may lead to health problems for human and other non-
target organisms in the environment (EPA, 2004). The use of pesticides for different purposes in
connection with contamination of environmental matrices and human exposure is illustrated in
figure 2.3.
16
Risk assessment of pesticide use in Ethiopia
Risk = exposure / effect
Pesticide use Ethiopia Exposure Effect
environmental
Synthesis
water Food: MRL
Formulation plant
Formulation soil Water: 0.1
Obsolete stocks air, food µg/l
Application
agricultural Greenhouse human
non-agricultural occupational
IRS applicator AOEL
Post-application worker ADI
food residues Illegal use
residential ARfD
environmental Deposition
consumer
Figure 2.3. Factors determining the contamination of environmental matrices, human exposure
and risks to pesticides
2.3. Pesticide residues in food
Even though, pesticides are important for agricultural productivity as well as to improve public
health, there is also a great public concern on human health risks (Wanwimolruk et al., 2015).
Pesticide used in agriculture have been detected in food and other environmental media, such as
in streams, rivers and sediments that are located near to agricultural sites which in turn may
aggravate human exposure (Dalvie et al., 2003). As a result, food stuffs such as cereals, fresh
vegetables, fruits and others, can be a potential source of pesticide uptake (Radwan and Salama,
2006).
Most of the time, consumers choose vegetables based on their attractive appearance, such as
proper size, colour and without damages by insects. Fresh fruits and vegetables are believed to
contain a lot of vitamins and minerals. To obtain this attractive appearance, farmers apply more
and more pesticides to control pests and diseases. However, the application of pesticides leaves
residues on the food items. From a study done in Ghana, different vegetables were contaminated
by different classes of pesticides (Bempah et al., 2012). Another study done in Thailand also
17
indicates that 25% of the vegetables purchased from market were contaminated with different
pesticides (Sapbamrer and Hongsibsong, 2014). Additionally, from a study in China, on the most
widely consumed vegetable in Asia (Chine’s Kala) reports that the residue of carbofuran,
chlorpyrifos, chlorothalonil, cypermethrin, dimethoate, metalaxyl and profenofos exceeded the
respective MRLs in 29% of the samples (Wanwimolruk et al., 2015).
In a study done in Nigeria, residues of two pesticides were detected in maize and up to 10% of
these pesticide residues were above the MRL (Ogah and Coker, 2012). Similarly, a study done in
Togo also describes that drinking water, maize, and cow pea were contaminated with
organochlorine pesticides such as HCH, DDT, aldrin, dieldrin, endrin, heptachlor, heptachlor-
epoxid, and endosulfan (α and β) (Mawussi et al., 2009). The other crop which possibly contains
pesticide residues is coffee beans. A coffee crop also suffers from diseases like fungi and others.
Application of pesticides often leads to a residue in the final coffee product. A fungicide,
flutriafol, was found in more than 30% of the green coffee beans samples in Brazil. A similar
study shows that there was a significant increase of coffee contamination by pesticides over the
years (de Oliveira et al., 2016).
Pesticides are also detected in food from animal sources such as milk. A study done on cow milk
collected from the market in Uganda reveals that organochlorine pesticides like lindane, aldrin,
dieldrin, endosulfan, DDT and its metabolites were detected mostly above the MRL (Kampire et
al., 2011). Chlorinated pesticides, mainly DDT and its metabolites were also detected in cheese,
yogurts and fresh milk collected from different communities in Ghana (Darko and Acquaah,
2008). In general, chemical pesticides are detected in raw agricultural crops and also in
processed foods in most developing countries (EFSA, 2009).
Contamination of food by pesticides potentially affects human health and food safety. The major
health concerns from exposure to pesticides are: cancer, damage to the reproductive system,
endocrine disruption, central nervous system, asthma and health problems in other body parts (
Ritter et al., 2006; Gilden et al., 2010).
18
2.4. Human exposure and toxicity of pesticides
Pesticides are developed through strict regulation to function with reasonable certainty and with
minimal impact on human health and the environment. However, major concerns have raised
about health risks due to occupational exposure, residues in food and in drinking water (Damalas
and Eleftherohorinos, 2011). Occupational exposure to pesticides happens during production,
transportation, mixing/loading and application of the pesticides in field ( Maroni et al., 2006;
Damalas and Eleftherohorinos, 2011). There are different factors that determine occupational
pesticide exposure such as frequency, duration, intensity, method of application and safety
behaviour, as well as the physicochemical and toxicological properties of pesticides (Hoppin et
al., 2006). In occupational exposure, the group of people at highest risk are persons working
directly with pesticides, family members of the pesticide applicators and agricultural workers.
These exposure arises from accidental spills of pesticides, leakages, incorrect use of equipment,
and non-compliance with safety guidelines (Fenske, 1997; Issa et al., 2010). Compared to
environmental exposure, occupational exposure is often at a relatively high dose, even if it is
short or long term exposure (Damalas and Eleftherohorinos, 2011).
The other route of exposure to pesticides for non-professional users is, through consumption of
contaminated food. This exposure depends on the choice of food and how much of that specific
food is eaten (Lu et al., 2006). Pesticide residues in food usually results from direct field
application (Enault et al., 2015). The climatic conditions in Africa promote the proliferation of
pests and weeds which results in a need for pesticide use which is considered as a major public
health concern among the African population (Nweke and Sanders III, 2009). Human beings are
on top of the food chain and it is estimated that over 90 percent, in particular chlorinated
pesticides, accumulate in the human body by food consumption (Center for Food Safety, 2006).
Exposure through air, drinking water and dermal contact is less. DDT accumulates in the fat
tissue of the human body and excretes via breast milk which may adversely affect the infants
health (Wong et al., 2005). Moreover, prenatal exposure to chlorinated pesticides has a potential
adverse effect on fetal development (Guo et al., 2014).
According to Renwick, (2002), the toxicity of pesticides can be evaluated at different time
frames and is classified mainly as acute (short term exposure) and chronic (long term exposure).
19
Intake of pesticides may not result in adverse health consequence in consumers unless it is taken
in excess. So, it is important to understand how high the dose of intake is and for how long a
person is exposed (Hamilton et al., 2004). Exposure to pesticide residues during one day time (24
hour exposure) is important to explain acute risk. The acute exposure can be substantially higher
than average because the consumption of food on a single occasion can be very large compared
with typical long-term consumption. Chronic exposure takes into account ‘non-consuming’ days
as part of producing an ‘average’ diet for long-term consumer risk assessment.
Organophosphates and carbamates (cholinesterase enzyme inhibitors) are the commonly reported
pesticides that cause acute pesticide poisoning and sometimes death (Colovic et al., 2013). The
acute effects are abdominal pain, headache, nausea, vomiting, diarrhoea etc. and the long term
effects such as cancer, depression, neurological deficits, genetic disorders, neurodevelopment
delays, endocrine disruption, respiratory distress, and impact on immune system (Andersson et
al., 2014). The chronic or long-term toxicity refers to adverse effects occurring as a result of
repeated exposure to an agent on a daily basis, or exposure to that agent over a large part of the
organism’s lifespan. The reference end point for acute toxicity is the acute reference dose
(ARfD) while, for chronic toxicity is the acceptable daily intake (ADI) (Solecki et al., 2005).
Once the human exposure level is above these two reference values, risk assessment should be an
important issue of concern.
2.5. Risk assessment
To determine the possible acute and/or chronic side-effects to a given pesticide residue from
consumer exposure, risk assessment needs to be done. As part of risk assessment, exposure
assessment of the pesticide residues found in the food is done by combining the amount of food
consumed and the amount of pesticide residues present in the food. For food safety evaluation,
the obtained exposure values are compared with ADI for chronic risk assessment and with ARfD
for acute risk assessment (Keikotlhaile et al., 2011). Risk assessment (RA) of a chemical is a
process intended to calculate or estimate the risk for a given target system following exposure to
a particular substance. RA takes into account the inherent characteristics of a substance of
concern as well as the characteristics of the specific target system. The risk assessment process
comprises four major steps figure 2.4.
20
Hazard Identification
Identify which health problems are caused by pesticide residues

in food
Exposure Assessment Dose-Response Assessment

Estimate dietary intake of pesticides from Evaluate the health problems from exposure
food, drinking water and khat to pesticides
Risk Characterization
Compare the estimated daily intake (EDI) with health based reference values (ADI and ARfD)
Figure 2.4. Risk assessment steps
2.5.1. Hazard identification
Hazard identification is the first step in the risk assessment process and it is the identification of
a risk source capable of causing adverse effect to humans or species in the environment, together
with a qualitative description of the nature of these effects. This activity is largely a qualitative
evaluation of the risk issue and preliminary examination of information that can be analysed in
the subsequent steps of the risk assessment. In toxicological risk assessment, the major focus of
hazard identification is to determine whether sufficient evidence exists to consider a chemical as
a cause of an adverse health effect (Lammerding, 2000).
2.5.2. Exposure assessment
Exposure assessment is one of the key elements in the risk assessment process. Exposure
assessment is defined as “the qualitative and/or quantitative evaluation of the likely intake of
biological, chemical, and physical agents via food as well as exposure from other sources”. It is
the estimation of how likely an individual or a population will be exposed to a chemical of
concern and how much of that chemical is taken up in the body through consumption of food,
21
drinking water and others (Lammerding, 2000). To assess exposure of chemicals in food, the
three most important aspects to be considered are: (i) how to determine quantitatively the
presence of a chemical in food; (ii) how to determine the consumption patterns of the individual
food items containing the relevant chemicals; and (iii) how to integrate both the likelihood of
consumers eating large amounts of the given food and the likelihood of the relevant chemical
being present in these foods (Kroes et al., 2002). The input data used in exposure assessment is
obtained from supervised field residue trials, national pesticide monitoring programs and food
consumption surveys. Exposure assessment can be done for acute or chronic exposures, where
acute exposure estimated for a period of 24 hours, while chronic exposure covers the average
daily exposure through the entire life.
2.5.3. Food consumption data
Food consumption data are the essential part to determine the dietary exposure to pesticides. The
data reflect which individuals or groups consume in terms of solid foods, beverages, including
drinking-water, and dietary supplements. There are four types of dietary assessment methods to
collect food consumption data, such as diet history, food frequency questionnaire (FFQ), dietary
records and dietary recall (EFSA, 2009). The quality of data from food consumption surveys
depends on the survey design, the data collection tools used, the motivation to respond and the
memory recall ability of the respondents. The methods that are suitable for both acute and
chronic risk assessment are dietary records and dietary recall. The most appropriate source is the
one that measures actual consumption instead of available food supply. Dietary recall is a
method used to collect consumption data, which involves asking the consumers to recall the
actual food intake for the past 24 or 48 hours or previous days. The most common dietary recall
method is the 24 hours recall. The data required for acute exposure assessment is, data on the
consumption of large portion size (for consumer population only) for a single consumption day,
for the edible part of the food and body weight of population who consume the food. While for
chronic exposure the non-consumer population are also included, considering the probability of
consuming through their life time (European Food Safety Authority, 2011).
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2.5.4. Dose-response assessment
The dose determines the poison, is a traditional dogma of toxicology. A dose-response

assessment is the determination of the relationship between the magnitude of exposure to a risk
source and the magnitude and frequency of the associated adverse effects. It analyses the total
amount of an agent absorbed by a group of population and the changes developed by reactions
with the body. As the dose increases, the measured response also increases. The dose effect
relationship for endocrine disrupting chemicals is different and the curve is non-monotonic i.e.
there is non-linear relationship between the dose and the effect. Endocrine disrupting chemicals
have an effect at low dose below the lowest dose at which a biological change for a specific
chemical has been measured before i.e. any dose below the lowest observed adverse effect level
(LOAEL). This challenges the traditional dogma of toxicology “the dose makes the poison” (
Vandenberg et al., 2012; Lagarde et al., 2015). Therefore, care should be taken in dealing about
the risk assessment of endocrine disrupting chemicals.
2.5.5. Risk characterization
Risk characterization is an integral component of the risk assessment process for both ecological
and human health. It’s an estimate of the probability of occurrence and severity of adverse effect
or event in a given population under defined exposure conditions. Or it is an integration of
evidence, reasoning and conclusions collected in hazard identification, dose-response assessment
and exposure assessment. It is the estimation of the probability, including attendant uncertainties,
of occurrence of an adverse effect if an agent is administered, taken or absorbed by a particular
organism or population. The potential risk is based on the assessment of end points such as the
ADI and ARfD derived from dose-response tests. If the risk is higher than these two assessment
end points, then a risk of exposure is happening (Hamilton and Crossley, 2004). The risk index
(RI) for human exposure can be calculated using the following formula:
RI = Exposure (Intake)/ Effect (ADI or ARfD)
Where:
RI = Risk Index
ADI = Acceptable Daily Intake, and
23
ARfD = Acute Reference Dose
2.5.6. Dietary exposure models for pesticide residues in food
Consumer risk assessment is a crucial step in the regulatory approval of pesticides on food crops
(Hamilton et al., 2004). The process of conducting dietary risk assessment involves the
determination of chemical residues in food or beverages and the calculation of the exposure
based on the consumption data for that specific food or beverages (Baker et al., 2001). To
quantify dietary exposure most often exposure models are applied which are important to support
dietary risk assessment. The dietary exposure models vary in complexity depending on the detail
of the exposure assessment. Deterministic and probabilistic models are the two important dietary
exposure assessment models.
2.5.6.1. Deterministic dietary exposure model
The deterministic exposure model can be used as a simple exposure modelling tool which relies
on fixed values (point estimates) derived from data. The deterministic calculation is done by
multiplying a fixed value of the food consumed and residue concentration, usually the mean or
97.5 percentile values (worst case scenario) (Kroes et al., 2002). Deterministic exposure models
are used as a low tier approach to determine whether there is an indication of concern for the
defined exposure. It makes part of the regulatory decision making guidelines due to their
simplicity, rapid and inexpensive character (EFSA, 2012a). The deterministic model does not
include information about variability in potential exposure to the exposed population. When
resource allows probabilistic assessment, it is preferred than deterministic assessment.
2.5.6.2. Probabilistic dietary exposure model
Probabilistic dietary exposure models are the proper representative of the true system. These
models take into account the distribution of one or more model parameters to represent real
variation and generate more realistic exposure estimates. Most of these distributional models are
based on Monte Carlo simulations and referred as Monte Carlo models (Hamilton et al., 2004).
These distributional models provide a range of risks throughout the population distribution and
24
provide quantitative information about variability and uncertainty. The simulation is repeated for
a certain number of iterations (e.g. 100,000) using statistical software such as @risk or Monte
Carlo Risk Assessment (MCRA) and results in an intake curve for the population of concern
(Kettler et al., 2015). However, the model requires time and resources for additional data
generation. In general, this model is important in risk assessment to assure food safety.
25
26
Chapter Three: Pesticide residue evaluation in major staple food items in Ethiopia
Chapter Three: Pesticide residue evaluation in major staple food items in

Ethiopia
Redrafted from: Mekonen S., Ambelu A. and Spanoghe P. (2014), Pesticide Residue Evaluation
In Major Staple Food Items in Ethiopia Using The QUECHERS Method: A Case Study From
The Jimma Zone, Environmental Toxicology and Chemistry, Vol.33. No.6. 1294-1302, DOI:
10.1002/etc.2554.
27
28
Abstract
Even though, the application of pesticides is important for protection of agricultural crops from
pests, their residues may remain in the crops as a contaminant. To determine the level of
contamination of the different commonly consumed food commodities (staple foods) in Ethiopia,
samples of maize (Zea mays), teff (Eragrostis tef), red pepper (Capsicum annuum), coffee
(Coffea arabica), lentil (Lens culinaris) and wheat (Triticum spp.) flour were collected from a
local market in Jimma zone, southwest Ethiopia. Samples were analysed for the presence of
cypermethrin, permethrin, deltamethrin, chlorpyrifos ethyl, DDT and its metabolites, and
endosulfan (α, β). For the analytical procedure, the Quick, Easy, Cheap, Effective, Rugged and
Safe (QuEChERS) extraction method with dispersive Solid Phase Extraction (d-SPE) clean up
technique was applied. The findings of this study indicate that the QuEChERS method is valid
for the analysis of the pesticides under study as the recoveries of most pesticides were in the
acceptable analytical range (70-120%) with a good repeatability (% RSD < 20). The limit of
detection (LOD) and limit of quantification (LOQ) varied between 0.001 to 0.092 µg/g and 0.002
to 0.307 µg/g, respectively. This designates that the method can fulfil the analytical requirements
for pesticide residue analysis. DDT, endosulfan, cypermethrin and permethrin were the major
pesticides detected in most food items. This is an indication of high utilization of these pesticides
in the study area. The concentration of the pesticides in the food items varied from 0.011 to
1.115 mg/kg food. All food items contained one or more pesticides and about one third of the
pesticides were above the maximum residue limit (MRL) set by Codex Alimentarius. Such
contamination is an indication of illegal use of pesticides in the study area. The organophosphate
pesticides such as malathion, diazinon and chlorpyrifos ethyl were detected in dehulled lentil and
wheat flour with a mean concentrations below the MRL set by Codex Alimentarius. The
concentration of most environmentally persistent pesticides such as DDT in coffee pulp was
significantly different (p-value < 0.01) from other food commodities except for red pepper. This
indicates that, red pepper and coffee pulp were highly contaminated by DDT. Therefore, there is
a need for a good pesticide monitoring program and evaluate food safety for the people in
Ethiopia.
29
3.1. Introduction
Wide spread application of pesticides for agriculture and disease control leave residues on crops
which may affect human health and the environment (Ogah and Coker, 2012). In areas where
there is widespread use of pesticides, food safety has become a number one public health
concern (Aktar et al., 2009). Pesticide residues can be found in different food items such as dairy
products, cereals, fruits, vegetables, cash crops like coffee and others (Lesueur et al., 2008;
Ahmed and Zaki, 2009; Keikotlhaile, and Spanoghe, 2011; Lozowicka et al., 2012; Bempah et
al., 2012; Gebremichael et al., 2013; Kapoor et al., 2013). In most developing countries the use
of chlorinated pesticides is common. For example in Nigeria, there is evidence of excessive use
of organochlorine pesticides (OCPs) by farmers in production of maize (Zea mays) (Tijani, and
Nurudeen, 2012). Due to this, about 96% of the maize samples contained residues of one or more
OCPs (Ogah, 2012). Additionally, in Ghana organophosphate, organochlorine and pyrethroid
pesticides were detected in vegetables and fruits (Bempah et al., 2012). Moreover, studies done
in Egypt, showed that cow milk was contaminated with OCPs (Ahmed and Zaki, 2009). In
addition to these, obsolete pesticides are documented as one of the major problems in Africa,
particularly in Ethiopia (Amera and Abate, 2008). OCPs are often detected in different food
commodities mainly due to environmental contamination from field application and leaching
from dumped obsolete stocks (Darko and Acquaah, 2008). In Ethiopia, indoor and outdoor
application of pesticides is a daily practice to increase productivity and to protect different food
items from various pests before and after harvesting. In addition, the Federal Ministry of Health
of Ethiopia is also applying indoor residual sprays (IRS) such as DDT and dieldrin to protect
people against mosquito transmitted diseases (Bekele et al., 2012). These applications potentially
contaminate different food items produced in Ethiopia and may have an impact on the health of
the public.
The most common food items produced in Ethiopia are cereals such as maize (Zea mays), teff
(Eragrostis tef), sorghum (Sorghum bicolor), millet (Pennisetum glaucum), coffee (coffea
arabica) and dairy products (Gebremedhin et al., 2007). Maize is one of the major crops
produced and a cheaper crop in Ethiopia. This may be the reason that maize is one of the major
staple food commodities for millions of people in the country, in particular to the Jimma zone
(Sori and Ayana, 2012). According to Gebremedhin et al. (2007), maize is the most important
30
crop in Ethiopia for household consumption (accounting for 65%) and covers about 47% of the
cultivated area in the country. In order to increase the yield of maize different hybrid varieties of
maize are cropped in Ethiopia. However, the hybrid varieties of maize can easily be affected by
different insects in the field and during storage after harvesting. As a result different pesticides
are applied and their residue may remain on the maize crop (Demissie et al., 2008).
Teff (Eragrostis tef), is primarily grown in Ethiopia and has been used for centuries as the
principal ingredient in the diet of Ethiopian people. The principal meal prepared from teff flour is
called injera which is a big flat bread or pancake, eaten alone or with any kind of sauces prepared
from meat, vegetables and other ingredients. In addition, teff flour is used for the preparation of
porridge and kita (non-fermented bread) (Hrušková, 2012).
Coffee (Coffea arabica), is also a popular cash crop in Ethiopia. Ethiopia is Africa’s biggest
coffee producer and exporter, followed by Ivory Coast and Uganda and it has been contributing
to more than 4% of the world coffee production (ICO, 2012). Along with production, coffee is
the most widely consumed stimulant beverage in the country as mentioned in chapter two. The
annual per capita consumption of coffee in Ethiopia is about 2.4 kg which is comparable to the
level of the leading coffee consuming countries (Zeru, 2006).
Red pepper (Capsicum annuum), is the other most ancient cultivated vegetable grown and
consumed in Ethiopia (Esayas et al., 2011). For most Ethiopians, food is considered as tasteless
without red pepper. The fine powder is an indispensable flavouring and colouring ingredient in a
common traditional sauce called “Wat” (Nsabiyera et al., 2012). Red pepper is the major spice
produced in more than nine districts of Jimma zone (Nsabiyera et al., 2012) and 50% of the
production goes directly to the local consumers (Shumeta, 2012). Therefore, if this vegetable is
contaminated by pesticides, there will be human exposure and a health risk can happen.
Ethiopia is also a major player in global production of legumes such as lentil (Lens culinaris) and
mostly devoted to produce red lentil. Lentil is the most important legume for human
consumption because of its high protein content (up to 35.5%) and Ethiopia produces lentil for
local consumption as well as for export (Sarker and Kumar, 2011). Lentil is important to prepare
a traditional sauce called “misir wot or lentil stews”. Before the preparations of misir wot, lentil
is semi-processed to dehull all the upper cover and is removed as dirt (personal observation).
31
Pesticides may be sprayed for control of lentil from pests and their residues may remain on the
crop. So analysing this type of crop is important for the safety of consumers.
Wheat (Triticum spp.) is also an important cereal in Ethiopia. It ranks fourth in area coverage
next to teff, maize and sorghum (Central Statistical Agency, 2008). Wheat flour is used for the
preparation of bread, porridge (locally called genfo), kita, traditional flat bread called injera and
local traditional beer called “tella”. Even though, wheat is highly produced and consumed in
Ethiopia; it may also be contaminated with pesticides sprayed for the control of weeds, insects
and other pests such as fungi, rodents and others. Daba et al. (2011), analysed wheat grain
collected from different regions of Ethiopia and detected organophosphate pesticides such as
malathion and diazinon and organochlorine such as aldrin and DDT. The detection of these
pesticides will have an effect on consumers’ health.
The agricultural office in Jimma zone reported that different types of pesticides are necessary to
guarantee a high productivity in the area. Pyrethroids (permethrin, cypermethrin and
deltamethrin), and organophosphates (chlorpyrifos ethyl, malathion, diazinon) are the major
pesticides applied in agricultural fields. Organochlorine pesticides (endosulfan and DDT) may
enter the environment by indoor residual spraying (IRS) for malaria control, from contaminated
surfaces at storage places and from the presence of obsolete pesticides in areas nearby (Mustefa
Temam, 2012, personal communication). The present study hypothesized that application of
these pesticides and historical use of some of the persistent pesticides such as DDT may results
in traces of residues in food items. The residues of pesticides can be found in all environmental
compartments however, the highest risk for consumers is through consumption of pesticide
contaminated food as 90% of residues, in particular organochlorine pesticides, accumulate in the
human body due to food consumption (Center for Food Safety, 2006; Price, 2008; Taha et al.,
2013).
From the accessible scientific literature in Ethiopia, there is almost no information regarding
pesticides residues in maize, teff, red pepper and coffee except, for wheat and khat (Daba et al.,
2011), cow and human milk (Gebremichael et al., 2013). This study presents data on the level of
pesticide residues in selected commonly consumed food commodities sold in the local market of
Jimma zone, southwest Ethiopia.
32
The quality of the analysis of the pesticide residues in food always depends on the available
analytical methods. Determination of pesticide residues in different food items using the
QuEChERS method is documented to give a better result and recovery compared to classical
techniques of liquid-liquid extraction (Document N° SANCO/12495/2011, 2011). The classical
extraction methods often need a large sample size because of multi-stage procedures and need
more clean up steps. As a result, classical methods are time consuming, labour intensive,
expensive and can produce a considerable amount of waste in the environment (Wilkowska and
Biziuk, 2011). Nowadays, analytical chemists prefer to use the QuEChERS method with a
dispersive solid phase extraction clean-up which is streamlined and effective for the analysis of
diverse residues in food matrices (Wilkowska and Biziuk, 2011). The QuEChERS method has
the following advantages: high recovery, high sample throughput, low solvent and glass ware
usage, less labour and bench space, lower reagent costs, ruggedness and low worker exposure
(Anastassiades et al., 2003; Lehotay et al., 2005). The aim of this study was also to evaluate
pesticide residues in selected staple food items of Ethiopia using the QuEChERS method as the
extraction and clean-up technique in the analysis.
3.2. Materials and Methods
3.2.1. Sampling
Samples of commonly consumed (staple food items) such as maize (Zea mays), teff (Eragrostis
tef), red pepper (Capsicum annuum) and coffee (Coffea arabica) were collected from the local
market in Jimma zone, southwest Ethiopia. This local market is a big market in the study area in
which most agricultural crops are collected for sale. During sample collection, the retailers in the
local market were interviewed about the origin of the crops and from how many farmers they
bought these crops. Most of the retailers mentioned that they bought one type of food item at
least from five farmers and pooled (not statistically) each food item before bringing to the market
for sale. Based on this experience, a retailer was chosen as a crop sample source, only if the
target crop was pooled at least from five farmers of a similar crop production location. Locations
where staple food items were commonly produced and coming to the local market were
mentioned in figure 3.1. In addition to these food items, semi-processed lentil (Lens culinaris)
and wheat (Triticum spp.) flour were also collected from the shops in the same market. This is
33
because; semi-processed foods such as dehulled lentil and ground wheat are usually sold in shops
rather than open markets. The collected samples each representing 250 g were sealed and
labelled with a unique sample identity and placed in a clean hard paper envelope within
polyethylene plastic bags. Then the samples were transported to the Laboratory and frozen at -20
0
C until the analysis was done.
Figure 3.1. Map of the study area indicating the food sample sources
3.2.2. Sample preparation
A 250 g of each sample was first ground and homogenized using a household mill equipped with
stainless steel knife (Fritel quality grinder OZX48-6cups) following the procedures used by
(Dasika, et al., 2012). After each sample was ground, the grinder was cleaned by washing to
avoid cross contamination. The ground and homogenized samples were packed in plastic bags
and stored at -20 °C until extraction was done.
3.2.3. Reagents and materials
Analytical grade acetonitrile was supplied by VWR-PROLABO and HPLC grade n-hexane and
acetone were obtained from ALLtech. Thermo Fisher Scientific supplied: magnesium sulphate
34
(MgSO4) to remove water from organic solvent, sodium acetate (NaAc) to absorb remaining
water content in samples, 50 mL polypropylene centrifuge tube, 15 mL dispersive solid phase
extraction tube (d-SPE tube) packed with primary secondary amines (PSA) for the removal of
organic acids and polar pigments among other compounds, and octadecyl (C18) to remove lipids
and sterols from the matrix. Pesticide standards, DDT (p,p’-DDE, p,p’-DDD, o,p’-DDT and p,p’-
DDT), endosulfan (α and β), cypermethrin, permethrin, deltamethrin and chlorpyrifos ethyl with
highest analytical purity were obtained from Supelco and delivered by Sigma-Aldrich logistic.
The respective purity of reagents, materials and pesticide standards are presented in table 3.1.
Table 3.1. Pesticides standards, reagents and QuEChERS materials with their respective
analytical purity
Pesticides standards Purity (%) Reagents Purity (%)
p,p’-DDE 99.9 Acetonitrile 99.9
p,p’-DDD 99.3 n-hexane 95
o,p’-DDT 100 Acetone 99
p,p’-DDT 99 glacial acetic acid 98.5
endosulfan α 98.5 MgSO4 98
endosulfan β 98 NaAC 99
deltamethrin 99 PSA 100
permethrin 98
cypermethrin 98
chlorpyrifos ethyl 99.5
MgSO4 = Magnesium Sulphate, NaAC = Sodium Acetate, PSA = Primary Secondary Amine,
C18 = Octedecyl
3.2.4. Analytical equipment
The pesticides were determined by gas-liquid chromatography with electron capture detector
(GC-ECD, Agilent Technologies 6890N) with an auto sampler. HP-5 capillary column of 30 m x
0.25 mm i.d. x 0.25 μm film thickness coated with 5% phenyl methyl siloxane (Model number
Agilent 19091J-433) was used in combination with the following oven temperature program:
initial temperature was 80°C, ramp at 30°C min-1 to 180°C, ramp at 3°C min-1 to 205°C, held for
4 min, ramp at 20°C min-1 to 290°C, held for 8 min, ramp at 50°C min-1 to 325°C. For
deltamethrin the oven temperature was maintained initially at 130°c, held for 1 min, ramp at
35
30°C min-1 to 280°C, held for 16 min and ramp at 50°C min-1 to 325°C, held for 3 min. The total
GC run time was 27.92 min. Helium (99.999% purity) was used as a carrier gas at a flow rate of
20 mL min-1 and nitrogen as a makeup gas at a flow rate of 60 mL min-1. An aliquot of 1µL was
injected in split mode at Split ratio of 50:1 and injection temperature of 280°C. The pesticide
residues were detected with electron capture detector (µ-ECD) operated at a temperature of
300°C. For the determination of pesticides, each food samples was analysed in triplicate and the
mean concentration was computed accordingly.
3.2.5. Analytical method validations: limit of detection (LOD), limit of quantification

(LOQ), repeatability (% RSD), linearity and recovery
The LOD and LOQ were determined by preparing matrix spikes at a low level near the expected
detection limit. The LOD and LOQ in this study were determined by spiking a different
concentration of the pesticides under study (chlorpyrifos ethyl, cypermethrin, permethrin,
deltamethrin, DDT and its metabolites (p,p’-DDE, p,p’-DDD, o,p’-DDT and p,p’-DDT), and
endosulfan (α and β)) in blank samples of maize, coffee beans, teff and red pepper in six
replicates. The LOD and LOQ were calculated by multiplying the standard deviation from the
replicates by 3 and 10, respectively (Butler et al., 2008). The accuracy and precision i.e., %
recovery and % RSD, respectively were determined with recovery experiments by spiking
pesticide standards in to laboratory blank sample of crops in six replicates with the same
concentration of each pesticide. Afterwards, the % RSD was calculated by dividing the standard
deviation with the average concentration. The recovery was calculated by dividing the recovered
concentration by the spiked concentration. The spiked samples were left for one hour before
extraction to allow the pesticides partition into the food matrices (Bempah et al., 2012).
The linearity was determined by preparing a stock solution of pure standards of the studied
pesticides and diluting them to produce different concentrations. The standard solutions of the
pesticides ran on GC-ECD under the set chromatographic conditions to produce five point
calibrations ranking from 0.01 to 1 mg/L. All the standard curves for each pesticides show, r2 >
0.995 which fulfil the requirements of good analytical method.
36
3.2.6. QuEChERS extraction method
The extraction and clean-up of the spiked samples and blank samples for each matrix was done
using the modified QuEChERS procedure with dispersive Solid Phase Extraction clean up (d-
SPE) method. The procedures were based on the AOAC official method 2007.01 (Lehotay,
2007) with slight modifications. This method gives high quality results for the analysis of many
pesticides in food (Dasika, et al., 2012). The Procedure for spiking and extraction is described as
follows: (i) 10 g of comminuted and homogenized blank sample of maize, coffee beans, teff and
red pepper was weighed in 50 mL centrifuge tube on an analytical balance (Sartorius AG): (ii) 10
mL of deionized water was added; (iii) blank samples were spiked with 25 µL of each pesticide
standard in each matrix in six replicates; (iv) 15 mL of acetonitrile (ACN) containing 1% glacial
acetic acid (v/v) in each sample was added using a solvent dispenser; (v) the tube was tightly
caped and shaken gently for 1 min to facilitate contact between the solvent and the sample; (vi) 6
g anhydrous MgSO4 and 1.5 g NaAc was added and the sample was shaken by hand vigorously
for 5 min to increase sample through put (Lehotay et al., 2005)); (vii) the sample was centrifuged
at 3000 rpm for 5 min.; (viii) to clean the extract, the upper organic layer was taken into a
dispersive solid phase extraction tube (d-SPE) containing 300 mg PSA, 900 mg MgSO4 and 150
mg C18, shaken by hand for 30 s and then centrifuged; (ix) a 5 mL aliquot of cleaned extract was
then taken and evaporated to dryness using a rotavapor (Rotary evaporator N18673, Switzerland)
at a temperature of 40°C; (x) the cleaned extract was reconstituted with 2 mL n-hexane/acetone
(9:1v/v) for solvent exchange; and (xi) then put into vial for GC analysis. The major procedures
are shown in figure 3.2.
37
Figure 3.2 QuEChERS extraction and clean-up procedures
3.2.7. Analysis of food samples
The extraction and clean-up of teff, maize, red pepper, coffee beans, coffee pulp, coffee beans
(after removal of the pulp), wheat flour and dehulled lentil, were done following similar
procedures for the above method validation tests (section 3.2.6) except spiking. From 250 g of
ground and homogenized samples, a representative 10 g subsample was weighed in a 50 mL
centrifuge tube on an analytical balance. Then, 10 mL of deionized water was added to the
sample and shaken for one minute. Then step iii-xi of the procedures was followed. The cleaning
procedure between samples was done by injecting a blank (n-hexane), pesticides standards for
calibration and n-hexane again to assure data quality. The sample equivalent (mg/mL) extract
was calculated based on the formula suggested by Schenck and Howard-King, (2000).
Y= a/b*x/z
Where:
Y= g sample equivalent/ mL of extract

a = g sample analysed
38
b = mL of solvent added to extract the sample

x = mL of the amount of the cleaned extract taken after the evaporation until dryness
z = mL of hexane/acetone (9:1v/v) added for solvent exchange
3.2.8. Data analysis
The wilcoxon signed rank sum test was performed to evaluate whether there is a significant
difference of pesticide residue between the food items. Box and whisker plots were used to
present the results. Statistical analysis considered significant at p-value < 0.05.
3.3. Results
3.3.1. Method validation results
The method validation results of the pesticides under study in replicate tests are shown in table
3.2. From our findings, the calibration curves obtained for a mixture of ten pesticide standards
including isomers and degradation products over a concentration range of 0.04-1 mg/L
(chlorpyrifos ethyl and cypermethrin), 0.05-1 mg/L for deltamethrin, 0.01-0.2 mg/L endosulfan
(α and β), 0.1-1 mg/L DDT metabolites (p,p’-DDE, p,p’-DDD, o,p’-DDT and p,p’-DDT) shows
a correlation coefficient (r2) > 0.995. This indicates the standard curves for all the pesticides are
clearly linear. The average recoveries of DDT and its metabolites, permethrin and chlorpyrifos
ethyl were in the acceptable analytical range (70-120%). However, the average recoveries of
cypermethrin were above 120% in all food matrices of the replicate tests, while, the most
lipophilic pesticide endosulfan both α and β isomers shows consistently low recoveries (< 60%)
but with good repeatability (% RSD < 10). This may be due to the physico-chemical properties
of these two pesticides such as their solubility in water and other organic solvents. LOD and
LOQ respectively varied from 0.001 to 0.092 µg/g and 0.002 to 0.307 µg/g for the pesticides
evaluated. The percent relative standard deviations (% RSD) are less than 11% for all the
pesticides. These results indicate that the method is accurate as most of the pesticides recovered
in the required analytical range (70-120%) and the method is also precise as the % RSD < 20.
39
Chapter Three: Pesticide Residue Evaluation in Major Staple Food Items in Ethiopia
Table 3.2. Method validation results of the evaluated of food matrices (maize, teff, red pepper and coffee beans)
Pesticides Spiked Maize Coffee beans Red pepper Teff
conc.
% LOD LOQ LOD LOQ LOD LOQ LOD LOQ
(µg/g) % Recovery % Recovery % Recovery
Recovery (µg/g) (µg/g) (µg/g) (µg/g) (µg/g) (µg/g) (µg/g) (µg/g)
chlorpyrifos ethyl 0.2 120.4 (1.4) 0.009 0.031 101.4 (1.0) 0.006 0.021 98.9 (1.6) 0.012 0.041 99.3 (1.6) 0.009 0.031
cypermethrin 0.2 143.2 (9.5) 0.059 0.197 121.7 (9.9) 0.032 0.107 132.2 (9.8) 0.061 0.202 135.1 (10.3) 0.062 0.207
permethrin 0.5 113.5 (4.9) 0.065 0.218 106.3 (8.8) 0.092 0.307 102.9 (4.7) 0.063 0.209 87.0 (5.7) 0.063 0.209
deltamethrin 0.5 123.3 (1.3) 0.022 0.075 135 (0.8) 0.016 0.055 108.9 (1.3) 0.018 0.06 96.9 (1.0) 0.015 0.05
p,p’-DDE 0.5 77.2 (1.0) 0.012 0.041 73 (1.4) 0.01 0.033 80.3 (1.2) 0.016 0.053 78.4 (1.1) 0.016 0.053
p,p’-DDD 0.5 102 (2) 0.028 0.092 80.1(1.7) 0.022 0.072 93.5 (1.5) 0.023 0.076 102.6 (1.5) 0.023 0.076
o,p’-DDT 0.5 86 (1.4) 0.015 0.051 86.7 (1.3) 0.015 0.051 89.4 (3.3) 0.045 0.151 94.1 (2.4) 0.036 0.121
p,p’-DDT 0.5 94.2 (1.6) 0.019 0.064 99 (1.8) 0.02 0.066 104.4 (2.5) 0.03 0.1 101.5 (2.2) 0.03 0.1
endosulfan α 0.05 57.4 (1.2) 0.001 0.002 50.1 (1.1) 0.001 0.004 50.6 (3.3) 0.004 0.014 52.7 (4.3) 0.004 0.014
endosulfan β 0.05 46.5 (1.7) 0.001 0.003 34.3(2.7) 0.001 0.004 44.1 (3.4) 0.005 0.016 38.1 (9) 0.004 0.014
Values in parenthesis indicate % RSD, LOD = limit of detection, LOQ = limit of quantification, % RSD = percent relative standard deviation
40
3.3.2. Pesticide residues in tested food samples
After validation of the QuEChERS method, the pesticide residues in maize, teff, coffee beans,
coffee beans with pulp and coffee beans after removal of pulp were determined. The results
reveal that each food item analysed had one or more pesticides figure 3.3. This indicates that the
environment in which these commonly consumed food items grown or stored are contaminated
by pesticides. Specifically, DDT, endosulfan, cypermethrin and permethrin, were the most
abundantly detected pesticides. The chlorinated pesticides such as DDT and endusulfan were
mostly used for malaria control in the study area. Their presence in the food items may be due to,
cross contamination or may be from their persistent nature in the environment. From the staple
food items, the red pepper and green coffee beans contained all the pesticides evaluated at
varying concentrations. Pesticide residues varied from 0.0321 to 1.148 mg/kg in red pepper,
0.011 to 0.301 mg/kg in maize, 0.014 to 0.351 mg/kg in teff, 0.016 to 1.115 mg/kg in green
coffee beans, 0.077 to 1.519 mg/kg in coffee pulp and 0.037 to 0.619 mg/kg in coffee bean after
removal of pulp. DDT and its metabolites; p,p’-DDE (19.04%), p,p’-DDD (92.9%), o,p’-DDT
(66.67%) and p,p’-DDT (100%) were detected in the food samples. The p,p’-DDT was detected
in all food samples compared to the metabolite p,p’-DDE, which indicates the recent use of the
pesticide DDT in the study area.
10 Chloropyrifos ethyl Cypermethrin DDT
Deltamethrin Permethrin Endosulfan
1
Concentration mg/kg
0.1
0.01
0.001
Pepper Maize Teff CoB CoPu CoBAPuR
Food samples
Figure 3.3. Average concentration (mg/kg) of pesticides in evaluated foods: Error bars indicate
the standard deviation; CoB = coffee beans; Copu = coffee pulp; CoBAPuR coffee beans after
removal of the pulp
41
Post hoc analyses compared the distribution of pesticides residues between crop samples and
illustrated by box and whisker plots figure 3.4. This comparison is important to identify which
food item is highly contaminated by specific pesticide and need food safety concern. For this
statistical comparison, the non-detects were included as being half the limit of detection (1/2
LOD) (Hornung and Reed, 1990). From the box and whiskers plot, DDT is highly distributed in
coffee pulp, deltamethrin in coffee beans and coffee pulp, chloropyrifos ethyl in red pepper and
coffee beans, cypermethrin and permethrin in red pepper, while endosulfan is highly distributed
in red pepper and coffee pulp. All food items contained one or more pesticides at varying
concentrations among which red pepper and coffee pulp were highly contaminated food items.
This raises the need for safety concerns for the general public consuming red pepper as major
food spices and rural Ethiopian community who mostly consume coffee pulp because it is
cheaper than the coffee beans (personal observation).
42
Figure 3.4. The concentration distribution of pesticide residues in evaluated food items. The
small square within each box represents the mean; the box represents the mean ± standard error;
and upper and lower whiskers represent 95% confidence intervals. Copu = coffee pulp; CoB =
coffee beans; CoBAPuR = coffee bean after removal of the pulp
43
3.3.3. Comparison of pesticide residue with the maximum residue limit (MRL) set by
Codex Alimentarius
Pesticide residues in red pepper, maize, green coffee beans, and teff were compared with
corresponding MRLs of each pesticide and indicated in table 3.4. Hence, there is no national
MRL for any pesticide used in Ethiopia, we consider the MRL set by Codex Alimentarius. From
all samples analysed, about two third had residues bellow the maximum residue limits (MRLs),
while about one third had a residue above the MRL set by the Codex Alimentarius (FAO/WHO,
2013). Cypermethrin exceeded the corresponding MRL (11.9%) most frequently, followed by
permethrin (9.52%), DDT (4.76%), and endosulfan, chlorpyrifos ethyl, and deltamethrin
(2.38%). The exceedance of MRL by the pesticide residues in the food items indicates that there
is illegal use of these pesticides in the study area.
Table 3.4. Comparison of pesticide residues (mg/kg) in food items with the maximum residue
limits (MRL)
Pesticides Red Pepper MRL Maize MRL Teff MRL* CoB MRL
chloropyrifos ethyl 0.032 2 0.011 0.05 ND 0.05 0.063 0.05
cypermethrin 0.390 0.1 0.156 0.05 0.351 0.03 0.553 0.05
DDT 0.734 NA 0.301 0.1 0.296 0.1 0.308 2.0
deltamethrin 0.069 0.03 ND 2.0 ND 2 0.440 1.0
permethrin 1.148 0.01 0.157 2.0 0.282 0.05 1.115 0.05
endosulfan 0.0599 5.0 0.012 NA 0.014 0.01 0.016 0.2
*
Values above the MRL are indicated in bold, The MRL of grains was adapted for teff, MRLs =
maximum residue limits; NA = not available; ND = not detected; CoB = coffee beans; CoPu =
coffee pulp; CoBPuR = coffee beans after removal of the pulp
3.3.4. Pesticide concentration (µg/kg) in lentil (Lens culinaris) and wheat (Triticum spp.)
flour
To see whether the semi-processed food items such as dehulled lentil and ground wheat powder
contain the residues of some pesticides, these two food items were collected from the market and
analysed. During screening of these two food samples ten pesticides (DDT, deltamethrin,
cypermethrin, permethrin, chlorpyrifos ethyl, pirimiphos methyl, diazinon, malathion,
fenpropimorph and 2,4-D) were checked. Seven of the screened pesticides were not detected
except malathion, diazinon and chlorpyrifos ethyl. The type of pesticides detected in dehulled
lentil and wheat flour were different from other food items such as teff, maize, coffee beans and
44
red pepper.This may be due to the source of the food items for the market. The retailers for teff,
red pepper, coffee and maize is brought these food items from the farmers in surrounding study
areas, while for dehulled lentil and wheat flour were coming to the market from different
factories. Therefore, the pesticides applied may be differed from areas to areas.
The organophosphate pesticides malathion, diazinon and chlorpyrifos ethyl were detected in both
lentil and wheat flour even though at low concentrations table 3.5. Chlorpyrifos ethyl is detected
in all lentil samples and in 86.7% of the wheat flour. Diazinon is detected in 50% and 83.3% of
the lentil and wheat flour samples, respectively. While, malathion was detected in all wheat flour
and 90.6% of lentil samples. The mean concentration of malathion and chlorpirifos ethyl were
far below the MRL in wheat flour and common beans set by the Codex Alimentarius
(FAO/WHO, 2013). As there is no maximum residue limit for semi-processed lentil MRL of
common beans was considered for comparison. The results indicate that the use of these
pesticides for the production of these two crops was with respect to the legal limit of the specific
pesticide. Or this can be explained as the semi-processing steps may decrease the pesticide
residue. Additionally, the organophosphate pesticides are not persistent in the environment, so
that the concentration in the food items may be decreased by degradations.
Table 3.5. Concentration of pesticides (µg/kg) of semi-processed lentil and wheat

Food items pesticides mean Min Max StDv MRL
diazinon 0.0012 0.0006 0.0142 0.0032 200*
Lentil malathion 0.0011 0.0002 0.0041 0.0009 2000*
chlorpyrifos 0.0019 0.0002 0.0121 0.0021 10 *
diazinon 0.0045 0.0006 0.0443 0.0085 N/A
Wheat flour malathion 0.0008 0.00001 0.0047 0.0012 200
chlorpyrifos 0.0009 0.0003 0.0049 0.0011 100
N/A = Not available, * = MRL of common beans
3.4. Discussion
The validation of the QuEChERS method indicates that the % recoveries for most of the
pesticides under study, the % RSD, LOD and LOQ were in agreement with the European
requirements for residue trials (Document N° SANCO/12495/2011, 2011). This points out that
the QuEChERS method is appropriate to determine pesticides in the corresponding food
45
matrices. The average recoveries of cypermethrin were consistently above 120% in all food
matrices of the replicate tests. If the recoveries of pesticides are repeatedly high or low in the
replicate tests, this outcome is acceptable according Document N° SANCO/12495/2011, (2011).
Among the food matrices, the recovery of endosulfan was lower in green coffee bean which was
50.1% and 34.3% for the α and β isomers of endosulfan, respectively. According to Pizzutti et al.
(2012) the modified QuEChERS method is known to obtain lower recoveries for lipophilic
pesticides such as endosulfan extracted from more fatty matrices like green coffee beans having
a 7-17% fat content. Both LOD and LOQ were below the lowest standard concentration which
indicates that the analytical method is able to detect and quantify lower concentrations from the
food matrices. The % RSD is the measure of agreement or the consistency of the analyte
concentrations to each other when the analyses are performed using identical conditions, i.e.
same method, same sample, same operator, and same laboratory conditions over a short period of
time (Geletu et al., 2009). The % RSDs for all the pesticides studied were below 11%, which is
in line with the commonly accepted analytical level (% RSD < 20) (Lehotay and Mastovska,
2005). The result of the present study is consistent with a study done in Portugal on multi-
pesticides residue analysis using acetonitril extraction, with a % RSD < 20 for all the pesticides
detected (Cunha and Fernandes, 2011).
From the box and whisker plot, a significantly higher concentration of cypermethrin was
observed in green coffee bean and coffee pulp relative to maize and teff (p-value < 0.01). This
indicates a high variation in contamination among food items. No significant differences were
observed for the concentration of chlorpyrifos ethyl between the evaluated food items. This
indicates that there may be a similar usage level of chlorpyrifos ethyl by farmers in the study
areas. The concentration of DDT in coffee pulp is significantly different (p-value < 0.01) from
other food items except for red pepper. This might be due to illegal use of DDT in coffee and red
pepper. As indicated in chapter two, DDT was used for malaria control in Ethiopia as indoor
residual spraying (IRS) ( Yewhalaw et al., 2011; Hamusse et al., 2012). There may be cross
contamination in the food items in particular for those food items stored in the IRS spraying
areas. According to Bempah et al. (2012), DDT is not degraded easily in the environment due to
its persistent nature, which may be also the reason for its presence in high concentrations in the
food items. In general, lower concentrations of pesticides were observed in samples of maize and
46
teff except for DDT which was above the MRL of grains set by Codex Alimentarious
(FAO/WHO, 2013). This may be due to a restricted use of the pesticides evaluated in these food
items except for DDT. The presence of DDT in the food items may also come from
environmental contaminations or there may also be illegally applied in agriculture. From the
coffee samples analysed, the maximum concentration of DDT was found in coffee pulp (1.52
mg/kg); while relatively lower concentration was found in coffee beans (0.30 mg/kg). The
concentration in coffee beans were lower compared to the coffee pulp, this might be due to the
rich nutrient content of the coffee beans, which may promote the growth of microorganisms that
are able to degrade the pesticides (Barragán-Huerta et al., 2007). Additionally, the maximum
concentration in coffee pulp may be due to the deposition of these contact pesticides (DDT and
its metabolites) on the surface of the crop as the pesticide is used not only during the growing
season but also during storage and transportation of the food items.
Compared to recent studies (from 2009 to 2012) in some African countries (table 3.6), the
commonly consumed food items in Jimma zone, southwest Ethiopia shows, higher
concentrations for all pesticide residues except the residues detected in food items in Nigeria for
DDT. This might be explained by the extensive application of pesticides in Ethiopia to obtain a
higher production, insufficient knowledge to apply the correct dose/rate and due to poor
equipment available to spray the pesticides (Amera and Abate, 2008). Additionally, higher
residues may result from historical use and previous environmental contamination, particularly
from those compounds demonstrating environmental persistence and accumulation of obsolete
pesticides near the study area (Amera and Abate, 2008).
The findings of our study in the analysed food items indicate a lower concentration of the
organochlorine pesticides than the study results observed in Nigeria. According to Tijani and
Nurudeen (2012), there is excessive use of organochlorine pesticides for the control of insects in
Nigeria. Although, DDT is officially banned for agricultural application, contamination of food
items and the environment still occurs. This contamination might be explained by drift from
indoor residual spraying (IRS) of DDT for malaria control or by leakage from dumped obsolete
pesticide stocks (Amera and Abate, 2008). Records indicate that the most persistent and
dangerous pesticides such as DDT, aldrin, heptachlor, pirimifos methyl and fenitrothion are the
main components in obsolete pesticide stocks which are dumped over 1000 sites in Ethiopia
47
(Hussien, 2007). Endosulfan was detected in 61.9% of the food items but in lower concentration
than the other pesticides studied. Compared to a study done in Togo, the residue in the present
study was three times and two times lower for endosulfan α and β, respectively. This might be
due to a lower contamination of the food items by endosulfan in our study area or the lower
recovery of this pesticide from the evaluated matrices.
Table 3.6. Comparison of pesticide residue in different food items of the present study with
findings in other African countries
Concentration
Country
Pesticides (mg/kg) Food items analysed Authors
p,p’-DDE 0.004-0.041 watermelon, cabbage, mango and banana
p,p’-DDT 0.004-0.038 carrot, watermelon, onion and banana
Ghana
chlorpyrifos 0.003-0.055 watermelon, tomato, pineapple or onion Bempah et

cypermethrin 0.008-0.035 mango, cucumber, papaya or tomato al, 2012
Permethrin 0.004-0.041 cucumber, pear, cabbage and pineapple
deltamethrin 0.004-0.044 carrot, pear, onion and pineapple
p,p’-DDE 0.001-0.175 chili pepper, tomato
Nigeri
Benson et
p,p’-DDD < LOD-0.172 chili pepper, tomato
a
al, 2011
p,p’-DDT 0.036-0.099 tomato, chili pepper
p,p’-DDE 0.005-0.007 cowpea, maize
p,p’-DDD < LOD-0.005 maize, cowpea
Togo
o,p’-DDT 0.002-0.055 maize, cowpea Mawussi

p,p’-DDT 0.011-0.023 cowpea, maize et al, 2009
endosulfan α 0.034-0.100 maize, cowpea
endosulfan β 0.065-0.090 maize, cowpea
chlorpyrifos 0.01–0.050 Wheat Dalvie and
Africa
South
cypermethrin 0.008–0.019 Wheat London,

Permethrin 0.014–0.448 Wheat 2009
p,p’-DDE < LOD-0.086 CB,CP, CBAPR, teff, red pepper and maize
p,p’-DDD 0.049 -0.128 maize, CBAPR, teff, CB, red pepper and Cp
o,p’-DDT 0.085-0.193 CB, CBAPR, CP, teff, maize and red pepper
Ethiopia
p,p’-DDT 0.099 -0.461 CBAPR,teff, maize, CB, red pepper and Cp

chlorpyrifos 0.011 - 0.063 CB or CP, teff, maize, red pepper and CB
This study
endosulfanα 0.004-0.033 CBAPR, teff, maize, CB, CP and red pepper
endosulfanβ < LOD-0.063 CBAPR, maize, CB, teff, red pepper and CP
cypermethrin 0.156 -0.553 maize, CBAPR, teff, CP, red pepper and CB
permethrin 0.157 -1.150 maize, teff, CP,CBAPR, red pepper and CB
deltamethrin 0.069 -0.157 maize or teff, red pepper, CBAPR, CP and CB
The food items listed according to the increasing order of pesticide residue, < LOD = less than limit of
detection, CB = Coffee Beans; CP = Coffee Pulp; CBAPR = Coffee Beans After Pulp removal.
48
Comparing the detected residues with the MRL it was found that DDT which was expressed as
the total sum of its metabolites: p,p’-DDE, p,p’-DDD, o,p’-DDT and p,p’-DDT, was above the
MRL set by Codex Alimentarius (FAO/WHO, 2013) in the two commonly consumed cereals
such as maize and teff. This indicates that there is illegal use of this pesticide in the production of
these two crops in study area and/or there may be contamination from the environment. Among
the metabolites, p,p’-DDE contributed less to the total DDT residue load. On the contrary, p,p’-
DDT which was found in all the food samples analysed contributed more to the total DDT
residue load. According to the European Food Safety Authority (Alexander et al., 2012), this
indicates that there is recent use of DDT in the study area. According to (Bempah et al., 2012),
even low exposure to pesticide residues of DDT accumulate and pose a health risk to consumers.
Consequently, under the framework of chronic and acute food safety risk assessments, pesticides
residue data below MRL should also be considered when assessing food safety (The 2010
European Union Report on Pesticide Residues in Food, European Food Safety Authority, 2013).
Semi-processed food items such as dehulled lentil and wheat flour also contain pesticide residues
such as malathion, diazinon and chlorpyrifos ethyl at a concentration range from 0.001 to 0.002
µg/kg and from 0.001 to 0.045 µg/kg in dehulled lentil and wheat flour, respectively. From a
study done in Ethiopia, diazinon and malathion were also detected in unprocessed wheat grain
collected from different regions of Ethiopia (Daba et al., 2011). The present study shows that all
the pesticides detected were below the MRL set by Codex Alimentarius (FAO/WHO, 2013).
This may be due to the semi-processing or may be due to the farmers may follow good
agricultural practice (GAP) during the production of wheat and lentil.
There were no MRLs found for the screened pesticides in teff and coffee pulp. This is perhaps
due to the unfamiliarity of these food items worldwide. For this study, the MRLs for grain were
considered to evaluate the residue in teff. As teff is the major grain consumed in Ethiopia and a
principal staple diet for almost all Ethiopians (Hrušková, 2012), setting an own MRL for the
different pesticides used on the crop is important. As the consumption of coffee pulp becomes
more and more important in rural population of Ethiopia (Kefale et al., 2012) monitoring of this
coffee component should also be a focus for consumer safety. Except for permethrin, the
concentration of all pesticides decreased in coffee beans after removal of the pulp. This might be
explained by the deposition of most pesticides on the surface of the crop. Pesticide application
49
can be undertaken at the pre-harvest season (during the growing season) or at post-harvest.
Hence, the pesticides may appear at the surface of the crop as a result of the time of application
or it may come from environmental contamination.
3.5. Conclusions
The QuEChERS method with dispersive solid Phase Extraction (d-SPE) clean-up was accurate
and precise for the analysis of the pesticides evaluated in commonly consumed food items. All
samples collected from the local market in Jimma zone, Ethiopia contained one or more
pesticides. This indicates there is contamination of the general environment, in particular to the
areas where the studied food items grown. Approximately, one third of the samples had pesticide
residues above the MRLs set by Codex Alimentarius, except for semi-processed lentil and wheat
flour. This indicates over use of pesticides in the southwestern Ethiopia. Hence, a great concern
has to be taken for the safety of consumers. Red pepper and coffee bean samples contained all
pesticides that were looked for. Some banned pesticides and those not authorized for use in
cereals, vegetables and coffee production such as organochlorines (e.g. DDT and endosulfan)
were also detected. This might be due to cross contamination during indoor residual spraying
(IRS) or due to the persistent nature of these pesticides, which make them to remain for a long
time in the environment. Recent use of DDT was also observed in the study area. This indicates
that farmers in southwestern Ethiopia use DDT illegally for the control of agricultural pests. An
establishment of national MRLs for pesticides used in the cultivation of these crops is necessary
to safeguard consumer health. More monitoring of pesticide residues in other food items will
result in a better risk characterization and assurance of food safety.
50
Chapter Four: Risk of DDT to infants consuming maize as complementary diet in southwestern
Ethiopia
Chapter Four: Risk of DDT to infants consuming maize as complementary

diet in southwestern Ethiopia
Redrafted from: Mekonen S., Lachat C., Ambelu A., Steurbaut W., Kolsteren P., Jacxsens L.,
Wondafrash M., Houbraken M., and Spanoghe P. (2015). Risk of DDT residue in maize
consumed by infants as complementary diet in southwest Ethiopia. Science of The Total
Environment, Vol.551, 554-560, http://dx.doi.org/10.1016/j.scitotenv.2014.12.087
51
Ethiopia
52
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Abstract
Infants in Ethiopia are consuming food items such as maize (Zea mays) in their complementary
diet. However, this may expose infants to toxic pesticide residues like DDT. A consumption
survey was undertaken using the 24 hour recall method for two non-consecutive days. Maize
samples were collected from the households visited during the consumption survey and from
markets in Jimma zone, southwestern Ethiopia. The residues of total DDT and its metabolites
were analysed using similar analytical method in chapter three. Deterministic and probabilistic
methods of analysis were applied to determine the consumer exposure of infants to total DDT.
The results from the exposure assessment were compared with the health based reference value
in this case the provisional tolerable daily intake (PTDI) set by the joint meeting of food and
agricultural organization and world health organization (JMPR of FAO/WHO). DDT was above
the detection limit in all maize samples, with a mean concentration of 1.77 mg/kg maize. The
mean concentration was far above the maximum residue limit (MRL) set by the Codex
Alimentarius. The mean and 97.5 percentile (p97.5) estimated daily intake (EDI) of total DDT
for consumer infants were respectively, 0.011 and 0.309 mg/kg bw/day from the deterministic
and 0.011 and 0.083 mg/kg bw/day from the probabilistic exposure assessments. For the total
infant population (consumers and non-consumers), the p97.5 EDI were 0.265 and 0.032 mg/kg
bw/day from the deterministic and probabilistic exposure assessment, respectively. Health risk
estimation reveals that the mean and p97.5 EDI of total DDT for actual consumers and p97.5 for
total population were above the PTDI set by FAO/WHO. Therefore, in Ethiopia, the use of maize
as complementary food for infants may pose a health risk due to the presence of DDT residue.
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Ethiopia
4.1. Introduction
Dichlorodiphenyltrichloroethane (DDT) was an effective insecticide until it was banned in most

industrialized countries since the late 1970’s. Even though it was banned, DDT is still used in
some African countries where malaria vector control is important (Bouwman and Kylin, 2009).
Although the use of DDT resulted in a successful elimination of the malaria vector (Beard,
2006), there is a great concern on its risks to human health (Eskenazi et al., 2009). DDT is a
persistent chemical in both biotic and abiotic environment, as a result its residue can be detected
in almost every human body (Vall et al., 2014).
Human beings can be exposed to DDT in utero, during breast feeding and through consumption
of contaminated food (Jusko et al., 2012). From all these routes of uptake, dietary intake is the
main route of human exposure because, 90% of the residue stored in a human body is due to
consumption of different food items (Center for Food Safety, 2006). Studies in different
countries indicate that food commodities are often contaminated with DDT. For example, a study
done in Turkey indicated that 83-96% of the wheat samples were contaminated with DDT and
its metabolites (Guler et al., 2010). Toteja et al. (2006) also report that residues of total DDT and
its metabolites were detected in 59% of wheat grains and in 78% of wheat flour in different
geographic regions of India. Additionally, a study done in Nigeria reports the presence of DDT
and its metabolites in maize (Tijani, and Nurudeen, 2012). Moreover, a recent study in Ethiopia,
also indicates that staple food items such as teff, maize, coffee beans and red pepper are
contaminated with DDT (Mekonen et al. 2014). In addition to dietary exposure, indoor residual
spraying (IRS) of pesticides for disease vector control may be another source of human exposure
(Yewhalaw et al., 2011). From a study done in South Africa, in indoor residual spraying (IRS)
villages, the level of DDT concentration in breast milk is higher than the MRL and the intake of
DDT by infants was above the PTDI (Bouwman et al., 2012). From the dietary exposure, the
actual intake may depend on the concentration of DDT and its metabolites in the food people eat
and the amount of food consumed (Agency for Toxic Substances and Disease Registry, 2002).
Compared to adults, infants are more susceptible to toxic pesticides like DDT when they are
exposed. This is because of their metabolic mechanism to detoxify these chemicals is not well
matured (Casals-Casas and Desvergne, 2011). Additionally, the infants food intake per body
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weight is higher compared to adults (Daston et al., 2003). In utero-exposure to organochlorine

pesticides like DDT has also a potential risk on the infants neurodevelopmental growth at the age
of 6-12 month (Eskenazi et al., 2006). Infants can also be exposed to DDT during breast milk
feeding. A study done in India indicated that the concentration of total DDT in breast milk for
some infants was above the tolerable daily intake (Bedi et al., 2013). Additionally, infants are
exposed to DDT when taking complementary homemade baby foods (Jeong et al., 2014).
In Ethiopia, maize is the dominant cereal used in complementary food administered to infants
after the age of 6 month (Akalu et al., 2010). However, maize can easily be affected by insects
in the field and during storage (Sori and Ayana, 2012). To tackle these insects, farmers use
pesticides including non-authorized ones, such as DDT which is actually banned for agricultural
use. A recent study done in Southwest Ethiopia indicates that DDT was found above the MRL
set by Codex Alimentarius in maize samples collected from Jimma zone, southwest Ethiopia
(Mekonen et al. 2014). If this crop is contaminated with DDT and its metabolites, infants are
exposed and face health risks. Pesticide application often results in residues in food which may
cause a health risk for humans (Akoto et al., 2013). So consumer risk assessment is an important
step for the regulation of pesticide use on food crops (Hamilton et al., 2004). Despite these
problems, there is no study that has estimated the exposure and risks of pesticides to infants,
particularly with DDT, in Ethiopia. The present study contemplated to investigate the risk of
total DDT for infants associated with consumption of maize in their complementary diets.
4.2.1. Study area
The study was conducted in southwest Ethiopia, around Jimma zone in the catchment area of
Gilgel Gibe hydroelectric reservoir, one of the biggest artificial lakes in Ethiopia (figure 4.1).
The study area includes four districts (Tiro Afeta, Sekoru, Omo-Nada and Kersa) around Gilgel
Gibe reservoir, which is located at 265 km southwest of Addis Ababa and 65 km north east of
Jimma town. Maize (Zea mays), sorghum (Sorghum bicolor) and teff (Eragrostis tef) are the
commonly grown food crops in the area. Among these cereals, maize production ranks first in
55
Ethiopia
Jimma zone (Gebremedhin et al., 2007) and used as commonly consumed food item in particular
to southwest Ethiopia (Sori and Ayana, 2012).
Figure 4.1. Map of the study area in southwestern Ethiopia
4.2.2. Dietary survey
To assess the food intake of 6-12 month old infants’ consumption data were collected for 130
infants from randomly selected households during the months of June to September. Socio-
demographic (age, sex and weight) data of the infants were collected during this household
survey. Dietary data was collected in a repeated 24 h recall interview administered by trained
interviewers using a face-to-face interview. This recall was done for two non-consecutive days
separated by 15 days. Food composition data, portion sizes, and recipes were entered and
processed in an online application to get dietary intake data (Lucille, Belgium,
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Ethiopia
www.foodintakesoftware.ugent.be). The amount of different ingredients of complementary foods

consumed by an infant per day was calculated. The average of both recall days was used to
estimate food consumption on a daily basis. The mean daily consumption (g/kg bw/day) for each
infant was calculated by dividing the infant’s average daily consumption (g/day) by his/her body
weight (kg). Weight of the infants was measured naked or with light clothes using mother-child
digital scales and recorded to the nearest 0.1 kg (SECA Uniscale, Hamburg, and Germany).
4.2.3. Maize sampling
A first batch of maize samples came from households interviewed during the dietary assessment.
In these households, the maize sampling was done randomly from different depths in the
traditional thatched basket stores. Assuming that the people, who may not have maize in their
stores, can buy from markets around the study area, a second batch of maize samples were
collected from four different markets (Sekoru, Omo-Nada, Kersa and Tiro Afeta). The maize
sampling from the market was done in similar way with Mekonen et al. (2014). Maize samples
(250 g each) were packed in polyethylene plastic bags, labelled accordingly and transported to
the laboratory. The samples were stored at -20°C until analysis.
4.2.4. Reagents and materials, analytical equipments, extraction and clean-up of the maize
samples, method validation procedures
The reagents and materials, the analytical equipment with the set chromatographic conditions,
extraction and clean-up of the maize samples and method validations were done according to
Mekonen et al. (2014).
4.2.5. Statistical and exposure analysis
The difference of the residue of total DDT between the market and the household maize samples
were evaluated based on the Mann-Whitney U test. As there was no statistical difference (p-
value > 0.05), all DDT residue data were consequently pooled and used for the dietary exposure
assessment of the infants. A Kruskal-Wallis test was performed to compare the residue of DDT
metabolites in the maize samples and a graphical representation was made using box and whisker
plot.
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Ethiopia
To evaluate the safety of consumers regarding pesticide residues, exposure should be assessed
and compared to toxicological limits or reference values (Claeys et al., 2011). For the present
study, the exposure assessment was done by assuming that the food processing (baking of maize
powder as a traditional Ethiopian flat bread (injera) or cooking in the form of porridge, before
administration to the infants, may not have an effect on the concentration of DDT (worst case
scenario). As recommended by the European Food Safety Authority, (2011a), dietary exposure
has to be done using deterministic and probabilistic approaches. The exposure assessment in the
present study was done for the actual maize consumer infants and for the total population (both
consumers and non-consumers). Including non-consumers was used to assess the chronic
exposure. The Mann-Whitney U test was applied to compare the estimated daily intake (EDI) of
total DDT by the infants from deterministic and probabilistic exposure analysis.
4.2.6. Deterministic exposure assessment
The dietary exposure to total DDT (mg/kg bw/day) was calculated deterministically based on the
maize consumption data (g/kg bw/day) and DDT residue (mg/kg maize). In this approach,
dietary exposure was computed by multiplying a single value of consumption and DDT
concentration (point estimate). The mean, p50, p75, p90, and p97.5 consumption was combined
with the mean, p50, p75, p90 and p97.5 total DDT concentration as explained by McKinlay et al.
(2008).
4.2.7. Probabilistic exposure assessment
A probabilistic exposure assessment was conducted using @Risk® 5.7 software program for
Microsoft excel 2010 (Palisade corporation, USA), in which the consumption (g/kg bw/day) and
residue distributions (mg/kg) are combined into an exposure distribution (mg/kg bw/day). First-
order Monte-Carlo simulation was undertaken considering 100,000 iterations. Estimated total
DDT intake (mean, standard deviation, maximum, minimum and percentiles) were determined
from the output of the simulation model.
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The results from the deterministic and probabilistic exposure assessment were compared with the
corresponding PTDI of DDT (0.01 mg/kg bw/day) set by JMPR of Food and Agricultural
Organization and World Health Organization (FAO and WHO, 2013).
4.3. Results
4.3.1. Socio demographic results
The percentage of male and female infants (6-12 month of age) was 61% and 39%, respectively.
The result indicates that more male infants are considered for the study, while in most of the
cases male: female ratio at birth is 1:1. This may be due to the fact that the consumption survey
was undertaken through a voluntary bases and mothers or care givers who have male infants may
be more volunteer than those who have female infants. The average weight of the infants was 7.6
± 1.25 kg. Regarding the maize consumption there is no significant difference observed among
the male and female infants (p-value = 0.354). This indicates that there is similar feeding pattern
for infants.
4.3.2. Concentration of total DDT and its metabolites in household and market samples
The distribution of total DDT from the two sample sources (households and markets) was not
significantly different (p-value = 0.321) as indicated in figure 4.2. This result revealed that the
contamination of maize samples with DDT was similar whether the infants consumed either
from household or from market maize sources. Additionally, the maize samples in the markets
may come from those households used for consumption survey because; the people use maize as
consumption commodity as well as income source.
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Ethiopia
Figure 4.2. The concentration distribution of total DDT (mg/kg) between household and market
maize sources. SE = Standard Error
DDT concentration in an increasing order for all maize samples is presented in figure 4.3. Total
DDT was above the LOD for all maize samples. This indicates that the study area is highly
contaminated by this very persistent pesticide in the environment. Most of the residue of DDT in
maize samples was above the MRL set by Codex Alimentarius (FAO/WHO, 2013). This
indicates that there is illegal use of DDT in the study area or the contamination might come from
the historical and/or the use for malaria vector control. The farmers or pesticide applicators in
Ethiopia usually stored the pesticides together with food commodities in the house or near to the
storage areas of the crops (personal observation). Due to these, cross contamination of the food
items may occur.
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Figure 4.3. Concentration of total DDT (mg/kg) in an increasing order of the maize samples.
LOD = limit of detection, MRL = maximum residue limit
The distribution of DDT metabolites in the maize samples is presented in figure 4.4. The result
of the present study revealed that p,p’-DDE, p,p’-DDD, o,p’-DDT and p,p’-DDT were detected
in 78%, 76%, 94% and 88% of the maize samples, respectively. From the box and whisker plot,
p,p’-DDT present in most of the samples followed by o,p’-DDT, while a very low concentration
of p,p’-DDE was observed. If the concentration of parent compound p,p’-DDT is higher than the
metabolite p,p’-DDE, then the result indicates recent use of DDT in the study area as has been
mentioned by European Food Safety Authority (Alexander et al., 2012).
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Ethiopia
4.0
Mean Mean±SE Mean±1.96*SE
3.5
(mg/kg )
3.0
2.5
Conc. (mg/kg)
Concentration
2.0
1.5
1.0
0.5
0.0
-0.5
DDD p,p’-DDE
p,p’- DDD DDE opDDT ppDDT
o,p’-DDT Total
p,p’-DDT DDTDDT
Total
l
DDTand
Total DDT metabolites
its metabolites
Figure 4.4. Concentration distribution of total DDT and its metabolites (mg/kg) in maize
samples. SE = Standard error
4.3.3. Exposure assessment
Deterministic and probabilistic analyses were worked out to evaluate whether the level of
exposure to infants to total DDT from maize consumption exceeds the PTDI set by FAO/WHO.
The mean and p97.5 of estimated daily intake were used to represent the average and high
consumer exposure scenario, respectively. A DDT intake above the PTDI indicates that there
will be a chronic health risk for the exposed infants. DDT has no acute reference dose (ARfD),
therefore, we did not consider the acute risk for this chemical.
4.3.3.1. Deterministic exposure assessment
The deterministic analysis was done based on the point estimate in which the mean maize
consumption was multiplied with the mean DDT concentration and the same is true for the
percentiles as indicated by McKinlay et al. (2008). Table 4.1 presents, the results of
deterministic exposure analysis to infants. The mean and p97.5 maize consumptions were 5.98
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Ethiopia
and 21.05 g/kg bw/day. The mean indicates the average consumption of maze by infants and
p97.5 indicates the high consumer group of infants’ i.e. if 97.5% of infants consume 21.05 g/kg
bw/day, the rest (2.5% of the infants) consume above this value. The mean and p97.5 total DDT
concentration were 1.770 and 14.700 mg/kg maize, respectively. Infants’ exposure assessment
indicates that the mean and p97.5 estimated daily intake (EDI) of total DDT were 0.011 and
0.309 mg/kg bw/day for actual consumers of maize, while 0.004 and 0.265 mg/kg bw/day for
total infant population. The median estimated daily intakes were 0.001 and 0.0001 mg/kg bw/day
for maize consumers and total population, respectively.
Table 4.1. DDT concentration, maize consumption and estimated daily intake (EDI) of total
DDT from the deterministic exposure assessment
Deterministic analysis Mean p50 p75 p90 p95 p97.5
DDT concentration
(mg/kg) 1.770 0.350 1.370 4.560 10.280 14.700
Consumption in consumer infants
(g/kg bw/day) 5.985 4.160 7.500 13.930 19.770 21.050
Consumption in total population*
(g/kg bw/day) 2.390 0.000 2.700 7.500 11.390 18.050
EDI of total DDT (mg/kg bw/day)
Consumers only 0.011 0.001 0.01 0.064 0.203 0.309
Total population* 0.004 0.0001 0.004 0.034 0.117 0.265
*including non-consumer infants, values in bold indicates results above PTDI = 0.01 mg/kg
bw/day
4.3.3.2. Probabilistic exposure assessment
In the probabilistic exposure analysis, both maize consumption and DDT residue data were used
in the @risk software to identify the best fit distribution using the Monte-Carlo simulation
model. From the fitting results, the P-P plot provided roughly a straight line joining the diagonals
for both consumption and DDT concentration. This is considered as the best fit distribution and
taken for simulation. During fitting of the distribution consumption for the total infant
population, data deviations from the normal line were observed. This deviation may be due to the
presence of zero consumption patterns. Table 4.2 presents, the distribution of the consumption
pattern, the distribution of total DDT concentration and EDI of total DDT by Ethiopian infants
from the probabilistic exposure analysis. The results reveal that the mean and p97.5 consumption
and DDT concentration were 6.484 and 30.153 g/kg bw/day; 1.770 and 13.155 mg/kg,
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Ethiopia
respectively. The mean and p97.5 estimated daily intake of total DDT were 0.011 and 0.083
mg/kg bw/day for consumers only; 0.004 and 0.032 mg/kg bw/day for total population. These
results are above the health based reference value of DDT (PTDI). This indicates that there will
be a health risk for the infants. The median (p50) estimated daily intakes were 0.002 and 0.001
mg/kg bw/day for consumers only and total population, respectively, which is below PTDI
similar to the deterministic analysis. This indicates 50% of the infants have no health risk from
consumption of maize in their complementary diet.
Table 4.2. Distribution in DDT concentration, maize consumption, and estimated daily intake
(EDI) of total DDT from the probabilistic exposure assessment
Probabilistic analysis Mean p50 p75 p90 p95 p97.5
DDT concentration (mg/kg) 1.77 0.415 1.29 4.015 7.771 13.155
Consumption among
Consumers (g/kg bw/day) 6.484 3.591 7.405 14.337 21.339 30.153
Consumption among total population
(g/kg bw/day) * 2.273 1.639 3.296 5.487 7.144 8.802
EDI of total DDT (mg/kg bw/day)
Consumers only 0.011 0.002 0.006 0.021 0.044 0.083
Total population* 0.004 0.001 0.002 0.008 0.017 0.032
*including non-consumer infants, values in bold indicates results above PTDI = 0.01 mg/kg
bw/day
4.3.4. Comparison of EDI of DDT for the total population in the deterministic and
probabilistic exposure assessment
The estimated daily intake of total DDT by the Ethiopian infants from the deterministic and
probabilistic exposure analysis was compared to see which method is appropriate to show the
best exposure scenario. Figure 4.5 revealed that the mean estimated daily intake of total DDT
for total infants from both deterministic and probabilistic exposure assessment were the same
and below the corresponding PTDI set by FAO/WHO. For the deterministic exposure assessment
the p90, p95 and p97.5, while for probabilistic exposure assessment only the p95 and p97.5
estimated daily intake (EDI) of total DDT were above the PTDI. This indicates that the
deterministic exposure analysis shows wider exposure scenarios. However, there is no statistical
significant difference between the two methods (p-value = 0.42).
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Figure 4.5. Comparison of estimated daily intake of DDT (mg/kg bw/day) from deterministic
and probabilistic exposure assessment for total infants. Y-axis in logarithmic scale
4.4. Discussion
There is no significant difference (p-value > 0.05) in consumption pattern of maize between the
male and female infant population. This may be due to the fact that maize is the commonly
consumed food item and similar feeding habits occur in each family in Jimma zone southwestern
Ethiopia. From the results it is found that there is no significant difference on the residue load of
DDT between market and household samples. This may be explained because of the reason that
the farmers who bring maize to the market are the same persons who have maize in their house.
Additionally, DDT contamination in maize may come from the same surrounding environment.
About three fourth of the maize samples had a total DDT residue above the MRL (0.1 mg/kg)
(FAO/WHO, 2013). The residue load above the MRL indicates that there is over use of DDT in
the study area for agricultural or household purpose. The detection of total DDT and its
metabolites in maize may also come from the application for indoor residual sprayings to control
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malaria (Yewhalaw et al., 2011). Additionally, its detection may be, explained by the persistent
or lipophilic nature of DDT as a contaminant in the environment (Bempah et al., 2012).
Moreover, DDT may come from environmental contamination with dumped obsolete pesticides
in different sites of Ethiopia (Amera and Abate, 2008). Among the metabolites of total DDT, the
concentration of o,p’-DDT and p,p’-DDT were significantly higher (p-value < 0.0001) than the
rest of the metabolites. However, no significant difference was observed in the concentration
between o,p’-DDT and p,p’-DDT, and p,p’-DDD and p,p’-DDE (p-value = 0.125). A similar
finding was observed in a previous study done in southwest Ethiopia (Mekonen et al., 2014).
The high concentration of p,p’-DDT indicates that there was recent use of DDT in the study
areas as has been explained by the European Food Safety Authority (Alexander et al., 2012).
From the deterministic exposure assessment it can be derived that the mean and p97.5 estimated
daily intake of total DDT for consumer infants were above the PTDI set by FAO/WHO, while
for the total infant population they were below the PTDI. The p97.5 estimated daily intake of
total DDT for the total infant population was also above the PTDI, while the mean estimated
daily intake of total DDT was less than PTDI.
From the probabilistic risk analysis, the upper percentile (p97.5) indicates that the infants were
exposed to high DDT concentrations. This result is consistent with the work of (Wason et al.,
2013) which reports that the upper percentile exposed children are exposed to a high
concentration of pesticides which influences their health more. Both exposure analyses indicate
health problems from consumption of maize as complementary diet.
Different studies explain that infants chronic exposure to organochlorine pesticides such as DDT,
develop health problems like attention deficient disorder, obesity and type two diabetics (Corin
and Weaver, 2005; Polańska et al., 2013).
From the comparison of the two exposure assessment models (deterministic and probabilistic),
the p90, p95 and p97.5 EDI of DDT for the total infant population from the deterministic
exposure assessment were above the PTDI while, for the probabilistic assessment it was only the
p95 and p97.5. According to the European Food Safety Authority (EFSA, 2012a) the
deterministic exposure assessment shows a wide range of exposure values as it uses a point
estimate for input and it generates a point estimate for exposure. However, the probabilistic
66
Ethiopia
exposure assessment uses distributions which take variability and uncertainties in the exposure
assessment into account. To see whether there is a significant difference in estimating the daily
exposure for the total infant population between deterministic and probabilistic exposure
assessment the Mann-Whitney U test was applied. The results revealed that there is no
significant difference (p-value = 0.42, U = 13, Z = 0.8) in estimating the daily intake of total
DDT for the total infant population from maize consumption even though the deterministic
analysis showed a wider range in exposure values. So both exposure assessment methods can be
appropriate here for the determination of exposure and consequently to assess the risk of the
infants to maize consumption.
4.5. Conclusions
This study estimated the exposure and risk of total DDT to infants from maize consumption in
their complementary diets. DDT was above the detection limit in all maize samples, with no
significant difference between market and household sample sources. About three fourth of the
maize samples contain DDT above the MRL set by Codex Alimentarius. This indicates that there
is illegal use of DDT in the production of maize or to protect storage pests and in addition to this,
the recent use of DDT was observed. The deterministic and probabilistic risk assessment
indicates that the average and high consumer estimated daily intake of total DDT for consumers
and total population were above the health based reference value. This may result in chronic
health problems for the infants. Monitoring of pesticides including DDT in other complementary
food items of infants is desirable. Furthermore, consumer risk assessment for the general
population in Ethiopia is important to contribute to the safety of human health.
67
Ethiopia
68
Chapter Five: Effect of household processing on pesticide residues in coffee (Coffea arabica)
and teff (Eragrostis tef)
Chapter Five: Effect of household processing on pesticide residues in coffee

(Coffea arabica) and teff (Eragrostis tef)
Part of this chapter is redrafted from: Mekonen S., Ambelu A. and Spanoghe P. (2015). Effect of
Household Coffee Processing on Pesticide Residues as a Means of Ensuring Consumers’ Safety.
J. Agric. Food Chem., 2015, 63 (38), pp 8568–8573, DOI: 10.1021/acs.jafc.5b03327
69
70
Abstract
Coffee (Coffea arabica) is a highly consumed and popular beverage all over the world; however,
coffee beans used for daily consumption may contain pesticide residues that may cause adverse
health effects to consumers. Teff (Eragrostis tef) is an endogenous cereal crop in Ethiopia and
also the most commonly consumed food item. Even though, the applications of pesticides such
as insecticides are not common in the production of teff, it may be contaminated from the
environment, leaking from storage areas and cross contamination during spraying for malaria
control. In this monitoring study, the effect of household processing on pesticide residues in
coffee and teff was investigated and the processing factor (PF) was calculated for each
processing method. Twelve pesticides, including metabolites and isomers (endosulfan α,
endosulfan β, cypermethrin, permethrin, deltamethrin, chlorpyrifos ethyl, heptachlor epoxide,
hexachlorobenzene, p,p’-DDE, p,p’-DDD, o,p’-DDT, and p,p’-DDT) were spiked in coffee
beans while, seven pesticides (permethrin, cypermetrin, chlorpyrifos ethyl, p,p’-DDE, p,p’-DDD,
o,p’-DDT, and p,p’-DDT were spiked in teff flour collected from a local market in southwestern
Ethiopia. The subsequent household coffee and teff processing methods such as, washing,
roasting, brewing, doughing, and baking were established as close as possible with the traditional
household processing methods in Ethiopia. From our findings, washing of coffee beans showes
14.6−57.7 percent reduction, while the roasting process reduced up to 99.8 percent. Chlorpyrifos
ethyl, permethrin, cypermethrin, endosulfan α and β in roasting and all of the twelve pesticides in
the coffee brewing processes were not detected. Kruskal−Wallis analysis indicates that the
reduction of pesticide residues by washing is significantly different from roasting and brewing
(p-value < 0.0001). However, there was no significant difference between coffee roasting and
brewing (p-value > 0.05). This indicates that roasting and brewing of coffee are the most
effective to decrease the residues of the pesticides under study. Dough preparation for teff flour
decreases the residues up to 86.4%, while baking reduces up to 90.2%. The PF was less than one
(PF < 1) for all the processing methods, which indicates reduction of pesticides under study
during household processing. The cumulative effect of these household processing methods are
important to evaluate the risks associated with ingestion of pesticide residues, particularly in
coffee beans and teff.
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5.1. Introduction
Wide applications of pesticides have different problems on human health and the environment as
explained in previous chapters of this thesis. Therefore, it is important for consumers to know the
possibilities to reduce pesticides intake from their food. As reported in literature, food processing
such as baking, cooking, roasting, peeling, washing drying etc. may reduce pesticide residues
from the raw food commodity (Kaushik et al., 2009).
Usually, pesticide residue analysis is undertaken on raw agricultural commodities to meet the
purpose of marketing to consumers, import/export certification, regulatory monitoring and
others. However, to estimate the level of exposure to pesticide residues in food, it is desirable to
investigate the level of pesticides at the point of consumption, mainly after food processing
which may lead to a reduction of pesticide residues (Satpathy et al., 2012).
Food processing is the action of transforming the food to a more edible form before the food is
consumed. The processing can influence the pesticide residue present after the raw agricultural
commodity is harvested (pranoy Pal and PG Sahah, 2008). Different household and industrial
food processing; such as washing with water or with different chemicals, peeling of fruits and
vegetables, frying, boiling, cooking and baking; may reduce the pesticide residue in food below
the risk level (Ahmed et al., 2011).
Coffee is a highly consumed and a popular beverage all over the world as explained in chapter
three. Coffee is the second most important commodity next to oil as a source of foreign exchange
for most producing countries. In addition, it is considered as a primary food, due to its contents
of compounds with an anti-oxidant effect and other beneficial biological properties. Its
characteristic flavour and aroma makes it a unique beverage with more than thousands of volatile
compounds present in roasted coffee (Yeretzian et al., 2003). As explained in chapter two,
Ethiopia is the top producer and exporter of coffee in Africa. The country is believed to be the
origin of Coffea arabica. On the other side, half of the coffee produced in Ethiopia is used for
local consumption which makes the country the leading African continent in coffee
consumption. Additionally, coffee is the most significant agricultural produce to develop the
country’s economy (ICO, 2012).
72
Specifically, the beautiful traditional coffee ceremony makes the country very unique in
consumption of coffee. The coffee ceremony and drinking of coffee is an important part of the
Ethiopian cultures. Coffee is offered during holydays, when visitors or friends come to home and
on a daily basis as a staple food item. Ethiopian coffee ceremony starts with washing the coffee
beans with water followed by roasting until they turn to a dark brown colour. After roasting,
sometimes the roasted coffee beans are brought to the family members or visitors to give them a
closer breath of the aroma. Afterwards, the beans are grinded using traditional wooden mortar
and pestle to obtain a fine powder. The powder is poured into boiled water in a special local
coffee clay pot called “jebena”. Coffee is ready to be served when the stem starts to come out
from the nozzle with an attractive flavour. After that the jebena is placed on the ground for about
three minutes, to let the coffee sludge settle to the bottom. The brewed coffee is then poured into
small cups and served (personal observation). However, the coffee beans used for this ceremony
may contain some hazardous pesticides.
Due to the high consumption of coffee and its economic importance, people from the producer,
exporter and consumer countries give more attention to its safety (Yang et al., 2011). Even
though, pesticide contamination in green coffee beans is limited during agricultural treatments,
there may be contamination during transportation and storage (Durand et al., 2006). According to
the investigation of (Jacobs and Yess, 1993), the imported green coffee beans samples to US
contained different pesticide residues such as chlorpyrifos at a concentration ranging from 0.01-
0.04 mg/kg and pirimiphos methyl at 0.01 mg/kg. From a study done in Brazil, residues of
different pesticides were detected in coffee beans either from registered or illegal use such as
endosulfan, chlorpyrifos, cypermethrin and captafol (Pizzutti et al., 2012). Additionally, from the
previous study done in Ethiopia, DDT, cypermethrin, permethrin, deltamethrin, chlorpyrifos
ethyl and endosulfan (α and β) were detected in coffee beans collected from the market in
southwest Ethiopia (Mekonen et al., 2014).
Teff (Eragrostis tef) is one of the major cereal crops, which is consumed by almost the whole
population in Ethiopia. Teff is the most important staple food crop, used to make injera, a
traditional fermented flat bread and for the preparation of porridge and kita (non-fermented
bread) in Ethiopia. Although teff is the preferred grain for making the staple injera (Yetneberk et
73
al., 2005), it gets little worldwide interest compared to other cereals such as rice, wheat and
maize.
Nowadays, the recognition that teff is gluten-free has spurred global research interest by
nutritionists and food scientists. Consequently, the number of studies on the nutritional
composition of teff and its health benefits increase (Baye, 2014). Teff is also the second most
important cash crop next to coffee, which generates about 500 million USD per year for local
farmers in Ethiopia (Minten et al., 2013) and is in the meantime an export commodity to many
European countries, Middle East and North America. It is a largely consumed crop especially in
urban and semi-urban Ethiopia with a national per capita consumption of 25.9 kg (Berhane et al.,
2011). Even though, this cereal is very important for human consumption and for generation of
income, it may be contaminated by different pesticides from application or from historical use.
As teff is not the common crop produced and consumed globally, there is almost no study done
on the status of pesticides residues in this grain. However, a recent study done in Ethiopia
detected cypermethrin, permetrin and deltamethrin, chlorpyrifos ethyl, and DDT and its
metabolites, in teff grain (Mekonen et al., 2014).
Pesticide residues, which are to a variable extent present in the food materials after harvesting,
are beyond the control of consumers. As Ethiopia is the largest producer, exporter and consumer
of coffee and teff in Africa, a pragmatic solution should be developed to tackle the problem of
coffee and teff safety. Different studies revealed that food processing such as washing, roasting,
cooking, baking, peeling drying etc. can be an important solution to reduce pesticide residues in
different food items ( pranoy Pal and PG Sahah, 2008;Ahmed et al., 2011; Keikotlhaile et al.,
2010;Satpathy et al., 2012).
Nowadays, there is an increasing need for information on the effect of various food processes on
pesticide residues. Few studies indicate an effect of coffee roasting on reduction of pesticide
residues (Cetinkaya et al., 1984; Sakamoto et al., 2012). However, no studies are available on the
effect of the household processing of coffee and teff such as washing, roasting and brewing,
doughing and baking methods on pesticide residues. Therefore, the main aim of the present study
is to evaluate the effect of household processing on pesticide residues in coffee beans and teff,
and to determine the processing factor (PF) for each of the processing steps.
74
5.2.1. Chemicals and reagents
Analytical grade acetonitrile (99.9% purity) were supplied by VWZ prolabo, HPLC grade n-
hexane (98% purity) and acetone (98.9% purity) obtained from ALLthec were used to extract
pesticide residues from raw and processed coffee beans and teff flour. Thermo Fisher Scientific
supplied MgSO4 (98% purity), NaAc (99% purity), 50mL centrifuge tube, 15mL dispersive solid
phase extraction clean-up tube packed with primary secondary amine (PSA) at 99% purity,
magnesium sulphate (MgSO4) and octadecyl (C18) at 99% purity. The pesticides under study
(p,p’-DDE (99.9%), p,p’-DDD (99.3%), o,p’-DDT (100%), p,p’-DDT (99%), endosulfan α
(98.5%), endosulfan β (98%), permethrin (98% ), cypermethrin (98%), deltamethrin (99%),
chlorpyrifos ethyl (99.5%), heptachlor epoxide (99.5%) and hexachlorobenzene (98%)) with
their highest analytical purity, were obtained from Supelco and delivered by Sigma-Aldrich
Logistic Analytical. Table 5.1 presents, the physico-chemical properties of the pesticides
investigated in this study.
Table 5.1. The physico-chemical properties of the pesticides under study

Water solubility Vapour pressure Mode of
Pesticides Logkow
(mg/L) (mmHg) action
p,p’-DDE 0.025 6.91 1.6*10-7 Contact
p,p’-DDD 0.120 6.51 1.4*10-6 Contact
o,p’-DDT 0.090 6.02 6.0*10-6 Contact
p,p’-DDT 0.085 6.79 1.1*10-7 Contact
endosulfan α 0.530 3.83 1*10-5 Contact
endosulfan β 0.280 3.62 1*10-5 Contact
permethrin 0.006 6.10 2.15*10-8 Contact
cypermethrin 0.009 5.40 2.3*10-7 Contact
deltamethrin 0.002 6.10 1.5*10-8 Contact
heptachlor epoxide 0.350 5.40 3.0*10-4 Contact
hexachlorobenzene 0.006 5.73 1.09*10-5 systemic
chlorpyrifos ethyl 1.400 4.70 1.9*10-5 Contact
Logkow = logarism of octanol-water partition coefficient
75
5.2.2. Sampling
A 3 kg of coffee beans (Coffea arabica) and 5 kg of teff (Eragrostis tef) from randomly selected
shops (1kg sample/shop) were bought from the local market in Jimma zone, southwestern
Ethiopia. These samples used as a blank and for spiking of the pesticides under study. Samples
were packed in polyethylene plastic bags after which they were sealed and labelled properly. The
samples were transported to the laboratory and stored at -20 °C until extraction was done.
5.2.3. Quality control
The quantitative determination of the pesticide residues in processed and unprocessed coffee
beans and teff were done based on an external standard method. The calibration curves were
obtained by injecting five different concentrations of the pesticide standards in the range of
0.005-1 mg/L. The regression coefficient (r2) of the standard curve was greater than 0.995 for all
the pesticides under study. Identification and quantification of the pesticides were done based on
the retention time and peak area, respectively.
5.2.4. Treatment of raw coffee beans
A 10 g raw coffee beans was weighed on the analytical balance and spiked with 40 µL of 100
mg/L of each pesticide under study in three replicates to increase the reliability of results.
Spiking was done for each household processing method for both coffee and teff (washing,
roasting, brewing, doughing and baking). The pesticides selected for this study were those
pesticides detected in the previous study on Ethiopian coffee beans and teff (Mekonen et al.,
2014). The coffee beans were sprayed on petri dishes to get equal distribution of the pesticides
for each bean. After spraying the petri dishes were covered with aluminum foil and placed in
refrigerator for 24 hours to increase the contact time between the pesticides and the matrix. After
one day, the processing methods (washing, roasting, brewing, doughing and baking) were
undertaken.
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5.2.4.1. Raw coffee beans
After one day the spiked samples of the raw coffee beans were grinded and homogenized using a
coffee grinder with a knife in it. The extraction and clean up of the raw and processed samples
were done using the same procedure in chapter three (Mekonen et al., 2014).
5.2.4.2. Washed coffee beans
The spiked coffee beans were washed thoroughly for 5 minutes under normal tap water (25-
30°C) resembling coffee bean washing at household levels. Then all other procedures in
unprocessed coffee beans were applied for the extraction, cleanup and analysis for the
determination of pesticides using gas chromathography with electron capture detector (GC-
ECD).
5.2.4.3. Roasted coffee beans
The spiked coffee beans were roasted at a temperature range of 230-240 °C (light to medium)
roasting and an average time of 12-14 min until the characteristic aroma or flavor of coffee
appeared. The roasting temperature and duration of the roasting was based on a study done by
(Moon and Shibamoto, 2009). Then the other procedures for extraction and cleanup used for raw
coffee beans were applied.
5.2.4.4. Brewed coffee
Following the roasting, the brewing process was undertaken. The roasted coffee beans were
grinded to fine powder using coffee grinder (Type 4041, 220-230V/150w). The fine coffee
powder was added in a coffee pot containing 100 mL boiled water and brewed for 10-12 min.
After brewing, the coffee pot was picked up from the stove and put on the ground until the
infusion of the coffee is cooled and the coffee sludge is settled down to the bottom of the coffee
pot. Then the upper liquid layer was taken carefully to the cups for further analysis. This brewed
coffee was extracted, cleaned up and analyzed in a similar way like that of the raw coffee beans.
77
5.2.4.5. Raw teff flour
A 5 g of teff flour was weighed on the analytical balance and spiked with 40µl of 100 mg/l of
each pesticide under study. After waiting for one day, the samples were extracted in a similar
procedure with the raw coffee beans.
5.2.4.6. Dough preparation
A 50 g of teff flour was weighed on an analytical balance and then spiked with 40µl of 100 mg/l
of each pesticide under study. The spiked teff flour was combined with 100 ml of water
(equivalent to 100 g water) and 0.5 g instant yeast, (together 150.5 g) and then mixed thoroughly
to get equal distribution of the pesticide throughout the dough. Then the prepared dough stayed
for 72 hours (three days). Then 15 g of the dough was weighed on an analytical balance using a
50 ml centrifuge tube. The extraction and clean-up was done in similar procedure as the raw teff
flour.
5.2.4.7. Baking
After sampling 15 g fermented dough for extraction, all the remaining dough was used for
baking of teff injera. The teff was backed using a pan in a similar condition with the traditional
Ethiopian injera baking. The baked injera was grinded and homogenized. Then from the
homogenous teff injera 10 g was taken and the extraction and cleanup of the samples was done
in similar way with the raw teff flour. During baking, the food is cooked in a closed environment
by heated air. Heat is supplied to the surface of the food by convention from the circulating air
and by conduction through the pan containing the food as explained in the report of (Fellows,
2009).
5.2.5. Determination of processing factor (PF)
The effect of household processing on the level of pesticides often correlates with the physico-
chemical properties of the pesticides under study, so it is important to adequately monitor the
processing factor. The PF for all transformation steps was calculated by the ratio between the
pesticide concentrations in the processed commodity (mg/kg) to the pesticide concentration in
78
the raw commodity (mg/kg). According to Bonnechere et al. (2012), a PF less than one indicates
that there is a reduction of pesticides by the processing method (PF < 1: reduction factor), while
a PF greater than one indicates no reduction in weight or volume, (PF > 1: concentration factor).
The processing factor is the proportional amount by which the residues changes when food is
processed. For this study PF was calculated by the formula below:
5.2.6. Analysis of wash water and coffee sludge
Before the coffee bean was roasted, it was washed with water. Additionally, after the coffee was
brewed there is sludge remaining in the bottom of the coffee pot. These two samples may contain
some pesticide residues that can be able to pass the washing and roasting process. Traditionally,
this wash water and coffee sludge is often disposed into the environment, which may
contaminate the foods grown there. To check whether the wash water from the samples and the
sludge of the coffee contain pesticide residues, these two samples were also analyzed for the
presence of pesticide residues. The procedure was as follows: a 10 mL of the wash water
(assuming 10 g of water) and 6- 8 g of coffee sludge were weighed on an analytical balance and
taken with a 50 mL centrifuge tube. Then the extraction, clean-up and analysis were done in a
similar way with the raw coffee beans.
5.2.7. Analytical equipment
Quantitative estimations and chromatographic separation of each pesticide were done by gas-
chromatography with electron capture detector (GC-ECD, Agilent Technologies 6890N) with an
auto sampler. The coffee bean processing was undertaken to see their effect on the pesticides
previously detected in Ethiopian green coffee beans (Mekonen et al., 2014), so that the
chromatographic conditions for this study were the same as in chapter three.
79
5.2.8. Statistical analysis
All the treatments of the processing methods (washing, roasting, brewing, doughing and baking)
were done in three replicates. Statistical significance was checked using Kruskal-Wallis test to
see if processing methods differed in reduction of pesticides under study. Differences at p-value
< 0.05 were considered as significant.
5.3. Results
5.3.1. Effect of household coffee processing on pesticide residues
This study investigated the effect of washing, roasting and brewing on the stability of twelve
pesticides. The concentration of the pesticides under study in raw and processed coffee beans is
presented in figure 5.1. Among the household processes, washing decreased the pesticide
residues in the range of 14.6- 57.7%. Maximum reduction was observed for endosulfan α and β
where the residue decreased to the extent of 53.1 - 57.7%, respectively. The results indicate that
the reduction of the pesticide residues after washing with tap water may be due to the removal of
some pesticides together with the thin upper layer of the coffee beans.
In this study, roasting is found to be effective to affect the stability of pesticide residues in
coffee beans. By this process, reduction of the pesticide residues was in the range of 72.5-99.8%.
Maximum reduction was observed for hexachlorobenzene (99.8%), followed by heptachlor
epoxide (97.9%), p,p’-DDE and p,p’-DDD (97.6% ) and o,p’-DDT (96% ). Five of the pesticides
(chlorpyrifos ethyl, permethrin, cypermethrin, endosulfan α & β) were not detected after the
roasting process. This may possibly due to instability of these pesticides due to the heat applied
in the roasting process as reported by other studies ( Bo and Zhao, 2010; Bajwa and Sandhu,
2014). After the coffee brewing process, none of the pesticides under study were detected. This
process is found to be the most effective to mitigate the pesticides under study in coffee beans.
80
0.7
Raw coffee bean Washed coffee bean Roasted coffee bean
0.6
0.5
Concentration (mg/kg)
0.4
0.3
0.2
0.1
Pesticides
Figure 5.1. Mean pesticide concentration (mg/kg) in raw and processed coffee. The error bars
indicated standard deviation
5.3.2. Determination of the processing factor (PF)
The PF is defined as the ratio of the residues in the processed food commodity to raw commodity
before processing. From the findings of our study, PFs for household processing of coffee was
less than one, which indicates that the processing methods have a reduction effect on the studied
pesticide residues. The PFs for the detected pesticides in the roasting process were the lowest
particularly for hexachlorobenzene, heptachlor epoxide, p,p’-DDE, p,p’-DDD, and o,p’-DDT
ranged from 0.001-0.04. This indicates that the roasting process played the most important role
in removing the pesticides from coffee beans compared to the washing process. The results
presented in table 5.2
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Table 5.2. The processing factor for each household processing method in coffee
Pesticide PF PF PF
Washing Roasting Brewing
endosulfan α 0.47 * *
endosulfan β 0.42 * *
chlorpyrifos ethyl 0.73 * *
permethrin 0.53 * *
cypermethrin 0.53 * *
deltamethrin 0.55 0.28 *
p,p’-DDE 0.73 0.02 *
p,p’-DDD 0.85 0.02 *
o,p’-DDT 0.72 0.04 *
p,p’-DDT 0.72 0.12 *
heptachlor epoxide 0.77 0.02 *
hexachlorobenzene 0.79 0.001 *
*for non-detected pesticides processing factor is not calculated
5.3.3. Pesticide residue in wash water and coffee sludge
Wash water after washing process and coffee sludge which were settled to the bottom of the
coffee pot after coffee brewing were analysed for the presence of those pesticides in raw coffee
beans. The result of the analyses reveales that all DDT metabolites (p,p’-DDE, p,p’-DDD, o,p’-
DDT and p,p’-DDT) and deltamethrin were detected in the wash water figure 5.2. This may be
due to the fact that these pesticides may be removed with the thin layer of the coffee beans
during the washing process. Additionally, the pesticide may come to the coffee beans from
environmental contamination and may appear on the surface of the beans can easily be removed.
Most of the pesticides disappeared from coffee beans during the roasting process. In the
meantime, pesticides were not detected in coffee sludge except deltamethrin and p,p’-DDT. The
detection of these two pesticides may be due to the lower percent reductions of deltamethrin
(72.5%) and p,p’-DDT (88%) during roasting process of coffee beans as a result, some part end
up to coffee sludge.
82
concentration in wash water concentration in coffee sludge

Average concentration of pesticides (mg/kg)
Pesticides detected in wash water and coffee sludge
Figure 5.2. Average concentrations of pesticides in wash water and coffee sludge
5.3.4. Effect of household processing on pesticide residues in teff
Figure 5.3 shows the effect of doughing and baking on pesticides residue in teff. The preparation
of dough decreases the pesticide residues in teff in the range of 59.9-86.3%, while baking
decreases in the range of 63.2-90.2%. This indicates that household processing has an effect on
reduction of the level of pesticides in teff and this will be helpful for the safety of consumers’
health.
83
1 Raw teff flour doughing baking

0.9
0.8
Average concentration (mg/kg)
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
Pesticides
Figure 5.3. The average concentration of pesticides in raw and processed teff, Error bar indicates
standard deviation
The processing factors for both doughing and baking were less than one, for all the pesticides
under study (table 5.3). These indicate that there is a reduction of the pesticide residue due to the
household processing of the teff flour. As reported in different literatures, PF < 1 indicates there
is reduction of pesticide residues due to processing ( pranoy Pal and PG Sahah, 2008; Hasmukh
et al., 2012; Bajwa and Sandhu, 2014).
Table 5.3.The processing factor for making dough and baking teff
Pesticide Raw teff flour PF Doughing PF Baking
chlorpyrifos ethyl 0.793 0.136 0.098
permethrin 0.793 0.129 0.099
cypermethrin 0.793 0.194 0.120
deltamethrin 0.793 0.267 0.154
p,p’-DDE 0.385 0.130 0.142
p,p’-DDD 0.657 0.092 0.112
o,p’-DDT 0.612 0.227 0.214
p,p’-DDT 0.551 0.401 0.367
PF = Processing factor
84
5.4. Discussion
Washing of coffee beans decreases pesticide residues up to 57.7%. This effect may be due to the
fact that most of the pesticides are found on the surface of the coffee beans. Surface residues are
removed by washing processes with the very thin layer of coffee beans located just above the
main coffee beans or with the removal of dust or soil on the coffee beans. Pesticides are applied
during the pre- and post-harvest stage of the crop. Contamination due to the presence of
pesticides takes place in both cases. In addition to this, pesticide contamination may come from
the environment due to previous persistent applications so that they may appear on the surface at
any stage of the crop. Indeed, (Ahmed et al., 2011) confirm that surface residues are removed by
washing processes with water. The majority of the pesticides applied on agricultural crops are
confined to the surface (contact action). Few pesticides really penetrate into the plant system
(Toker and Bayιndιrlι, 2003). As a result, they are removed by washing, trimming or peeling.
During the washing process, a maximum reduction was observed for α- and β-endosulfan for
which the residue decreased to an extent of 53.1 to 57.7%, respectively. A similar result was
observed from a study done in India. Washing of the vegetable brinjal (an eggplant) with tap
water decreased endosulfan residues up to 55%. The researcher concluded in that rinseability of
some pesticides by washing with water not always correlates with their water solubility (Cengiz
et al., 2007). As indicated in table 5.1, most of the pesticides in this study have low water
solubility with a high octanol-water partition co-efficient (logKow > 3). As a result washing was
comparatively found less effective in reducing the residues of the lipophilic pesticides such as
p,p’-DDD (14.6%), hexachlorobenzene (20.8%), heptachlor epoxide (23.4%), chlorpyrifos ethyl
(26.5%), o,p’-DDT (28%) and p,p’-DDT (28%). (Yoshida et al., 1992), reports that heptachlor
epoxide in cucumber was not well removed during the washing process.
In this study, roasting is found to be effective to affect the pesticide residues in coffee beans.
This process reduces the residue in the range of 72.5 - 99.8%. Thermal processing treatments like
cooking, baking, blanching, steaming, roasting and boiling have been found effective for the
dissipation of various pesticides (Bajwa and Sandhu, 2014). Additionally, there is a loss of
pesticides due to the heat treatment in related to the physico-chemical properties of the pesticides
(Sharma et al., 2005). A study done in Japan on the behaviour of pesticides in coffee beans
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reports that during the roasting process the pesticide residues decrease to a significantly lower
concentration (Sakamoto et al., 2012).
After the coffee brewing process, none of the pesticides under study were detected reflecting a
100% removal. This process is found to be the most effective method to mitigate the problem
related to pesticides under study. As it has been explained by (Abou-Arab and Abou Donia,
2001) the elimination of pesticide residues from the boiled extract of coffee may be due to the
decomposition of the pesticide residues by the application of heat. The pesticide residues were
significantly influenced by the roasting and brewing processes. In general, the three household
coffee processing methods have a cumulative effect in the reduction of pesticides as a result the
safety of consumers is assured. Figure 5.4 illustrates the cumulative reduction of the pesticides
by the household processing methods.
Figure 5.4. The percent reduction of pesticide residues during household processing of coffee
The Kruskal-Wallis test showed that there is strong variation in the stability of pesticide residues
in coffee beans among the processing methods (p-value = 0.00001). The reduction of pesticide
residues due to washing is significantly lower compared to roasting (p-value = 0.0001) and
brewing processes (p-value = 0.00001). According to Hasmukh et al. (2012) the washing process
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depends on different factors like location, age of residue, water solubility and temperature which
explains the variation. However, there is no significant difference between the roasting and
brewing process (p-value > 0.05). As indicated in the figure 5.4, the roasting process decreases
the residues of the pesticides in this study up to 99.8% ~ 100% just like coffee brewing. This
may be due to the application of heat in both roasting and brewing. Heat affects the stability of
the pesticides (Bajwa and Sandhu, 2014). Pesticide residues were also determined in wash water
and coffee sludge. The results indicate that all DDT metabolites and deltamethrin were detected
in the wash water while only p,p’-DDT and deltamethrin were detected in coffee sludge. The
detection of p,p’-DDT and deltamethrin in coffee sludge may be due to the lower percent
reduction of these compounds during the roasting process compared to the other pesticides. The
analysis of the residue of these two pesticides in the coffee sludge is important because, in
Ethiopia, the wash water from the coffee beans and the sludge after brewing the coffee are
discarded into the open environment which may indirectly contaminate the food. So, care should
be taken during the disposal of both the liquid (wash water) and semi solid (coffee sludge) waste.
In addition, during pouring of the coffee solution to drink or serve coffee, only the upper layer of
the coffee infusion should be drunk for the safety of consumers.
The household processing of teff to dough and baking of teff injera also decreases the pesticide
residues under study. Dough preparation decreases the residue of the pesticides up to 86.4%.
This may be due to the fermentation process for three days. Fermentation is a process which
leads to a large reduction in pesticide residues from the original amount (Regueiro et al., 2015).
Sharma et al. (2005) also reports that fermentation facilitates the reduction of pesticides in food
products.
The baking process decreases the residues in particular chlorpyrifos ethyl up to 90.2%. This may
be due to the application of heat during the baking process. According to Byrne and Pinkerton,
(2004), baking decreases the residue of chlorpyrifos up to 98% in winter squash and up to 70%
in sweet potato.
The preparation of dough and baking has a lower removal effect on p,p’-DDT (parent compound
DDT). The residue of p,p’-DDT reduced only up to 59.9% and 63.3% during doughing and
baking, respectively. This may be due to the persistent nature of DDT for degradation by bacteria
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during fermentation or resistance to heat application during baking. The processing factor (PF)
for both coffee beans and teff flour were below one for each processing steps and indicates a
reduction which is also reported by (Bonnechere et al., 2012).
5.5. Conclusions
Food is a basic need to human beings more than shelters and/or clothing. However, food may be
contaminated by different chemical, physical or biological hazards. Pesticides are one of the
major chemical hazards in food. Different scientific investigations are required to understand the
real dietary intake of pesticides to setup regulatory standards in commonly consumed food
commodities. To support these processing studies, it is important to understand the real dietary
intake of pesticides. The present study demonstrates the dissipation of the pesticides under study
during household processing of coffee and teff. Roasting and brewing of coffee were the most
effective methods to reduce pesticide residues. Pesticides were detected in coffee sludge. Hence,
the mix up of the coffee with the residue should be avoided and the brewed coffee infusion
should be mandatory to drink pesticide free coffee. The household teff processing has also an
effect on the reduction of pesticide residues. Good reduction of the residue was observed during
the baking process. This may be due to the application of heat. The PF for both coffee and teff
were less than one (PF < 1) which means that there is a reduction of pesticides due to the applied
household processing methods. Such reduction is important in evaluating the risks associated
with ingestion of pesticides, specifically in coffee which is a widely consumed beverage in the
world and teff which is the most commonly consumed cereal in Ethiopia. The result of this study
assured that household processing of coffee and teff have a paramount effect on the reduction of
pesticide residues and this initiates to work on the effect of different household processing
methods on other pesticides.
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Chapter Six: Pesticide residues in drinking water and associated risk to consumers in Ethiopia
Chapter Six: Pesticide residues in drinking water and associated risks to

consumers in Ethiopia
Redrafted from: Mekonen S., Argaw R., Simanesew A., Houbraken M., Senaeve D., Ambelu A.
and Spanoghe P (2016). Pesticide residues in drinking water and associated risk to consumers in
Ethiopia. Chemosphere, 162, 252-260, doi.org/10.1016/j.chemosphere.2016.07.096
89
90
Abstract
Access to safe and reliable drinking water is vital for a healthy population. However, water
sources may become largely contaminated with pesticides because most of them are located
nearby agricultural areas. Water samples were collected from different water sources (river,
spring, well, pond, and community taps) in Jimma zone and Addis Ababa, Ethiopia. The
extraction and clean-up of the samples were undertaken using liquid-solid and liquid-liquid
extraction methods. Human exposure to the detected pesticides was assessed by calculating the
estimated daily intake (EDI) of pesticides from the water. To evaluate the risk, the EDI was
compared with the acceptable daily intake (ADI) for the long term exposure and the acute
reference dose (ARfD) for the short term exposure. The findings of our study indicate that 2,4-D,
malathion, diazinon and fenpropimorph were present in all water sources while pirimiphos
methyl was detected only in well and spring water. The mean concentration of 2,4-D, diazinon,
malathion and fenpropimorph from Jimma water sources, were in the range of 2.27 to 2.94, 1.59
to 5.65, 1.71 to 5.03 and 7.25 to 13.90 µg/l, respectively. Only 2,4-D, diazinon and
fenpropimorph were detected in the water source of Addis Ababa with a mean concentration
range of 0.60 to 69.10, non-detected to 32.10 and 0.11 to 138 µg/l. From both sample sources
(Jimma and Addis Ababa), the residues of some of the pesticides were above the European
drinking water guide line set by European Commission (EU) and World Health Organization
(WHO), which indicates that there is overuse of pesticides by farmers in the study area.
Concerning human health risk estimation, all the pesticide detected have no acute risk for
consumers (EDI < ARfD). However, chronic risks to human health were observed from
diazinon and fenpropimorph (EDI > ADI), for Jimma and Addis Ababa population. A
comprehensive monitoring and source oriented remediation action is required to reduce the level
of pesticide residues in water and to minimize particularly the long term health risks on human
health.
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6.1 Introduction
Access to clean water is a fundamental human right and vital to sustain healthy life. Reports,
however, indicate that residues of pesticides occur in different water sources ( Ntow, 2001;
Sankararamakrishnan et al., 2005; Flores-García et al., 2011; Chowdhury et al., 2012; Varca,
2012). Environmental contamination of natural water by pesticide residues during and after field
applications is of a great concern. Water is an important component of public health and failure
to supply safe drinking water causes a heavy health burden to humanity (Kooij, 2014).
The contamination of surface and ground water by pesticides is discussed around the world
(Donald et al., 2007; Varca, 2012). A study done by Teklu et al. (2015), detected 2,4-D,
malathion, deltamethrin, atrazine, chlorothalonil and endosulfan in surface water in Ethiopia.
Pesticides may enter into the aquatic system by diffusion to the surface or subsurface
hydrological pathways. Pesticide transport to surface water, is mainly caused by spray drift,
runoff water and drainage water (Ikehata and Gamal El-Din, 2005). This may happen, due to
improper operations such as filling of sprayers, washing of measuring utilities, disposing of
packing materials and cleaning of spraying equipment. The pesticide sprayers in Ethiopia can do
mixing/loading or dilution of pesticides near to water sources, as illustrated in figure 6.1, which
contaminates the water as well as the irrigated crops. In turn, such practices affect the health of
the communities living around these areas. Additionally, applications on lawns and runoff during
rain events are prone to be flushed into the sewerage channels and end up in the receiving water
bodies (Gerecke et al., 2002).
Due to the persistence of some pesticides in the environment and potential adverse acute or
chronic health effects for consumers, contamination of surface and ground water has long been
recognized as an important issue in many countries. Understanding the fate and quantity of
pesticides before and after treatment of drinking water is crucial (Benotti et al., 2009). In
developing countries, the need to ensure local agricultural production and food security while
simultaneously protecting the population against health effects from pesticide exposure remains
a major public health challenge. According to Kesavachandran et al. (2009), developing
countries use only a small amount of the world’s agrochemicals, but they suffer from 99% of
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deaths due to unsafe application of the pesticides and poor handling due to illiteracy and poverty
status of the users.
Figure 6.1. A farmer preparing pesticides not far from a water source which is used by the
communities living around (Environment and Social Assessment International, 2006)
Although Ethiopia has a huge potential of surface and ground water sources, the country utilizes
a small portion of these resources. Nowadays, the access to safe drinking water is a bit improved.
According to central statistics agency of Ethiopia, more than half of the households in Ethiopia
(57%) have access to an improved source of drinking water, with a much higher proportion
among urban households (Central Statistical Agency [Ethiopia], 2014).
Some of the water sources in the rural community around Jimma zone, southwest Ethiopia are
springs, well water, and ponds. The dwellers of Jimma town use water from different sources
such as tap water, springs, wells and rivers for drinking and other domestic purposes. There is
one conventional water treatment plant in the town which supplies purified water for the
population of the town (Kifle and Gadisa, 2006). Other parts of the communities in Jimma zone
do not have a conventional water treatment plant but rely on the different water sources such as
springs, well and river water.
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Addis Ababa, the capital of Ethiopia and the diplomatic centre of Africa, is one of the fastest
growing cities in the continent. Its population has nearly doubled every decade. The city is
subdivided into ten sub-cities with a total population of 3,048,631. The main water supply
sources in Addis Ababa are generated from the three ponds (Geferesa, Legedadi, and Dire) and
ground water source (Akaki ground water). Water coverage of the city is about 94% in 2012
with a daily supply of 374,000 cubic meters (City Government of Addis Ababa, 2013).
The Legedadi pond is the largest water supply source which contributes 40% of the town water.
The water treatment capacity of the plant is 150,000 cubic meters per day (UNEP/UNESCO/UN-
HABITAT/ECA, 2003). The catchment area of Legedadi is about 206 square kilometres and is
surrounded by farm lands. These farm lands are owned by smallholder farmers who use different
crop protection products. During application these pesticides may contaminate the environment
in particular the water sources which are used for drinking by the communities living around.
The other possible sources of pesticide contamination in the drinking water sources may be
attributed to leaks of pesticides from obsolete stockpiles near the study area (''Sireguyo pesticides
store'') (personal observation). Inappropriate storage, poor handing practice, inappropriate
labelling (labels are in English rather than in local language), and illegal use of pesticides due to
poor knowledge of farmers may aggravate the contamination of the water sources (Haylamicheal
and Dalvie, 2009).
The Jimma and Legedadi water treatment plants use conventional water cleaning processes, such
as coagulation/flocculation, filtration and chlorination which have been shown to be fairly
ineffective to completely remove pesticides (Saifuddin et al., 2011). Recent studies indicate that
reverse osmosis, nano-filtration and adsorption on granular and powdered activated carbon are
more effective to remove pesticides (Benotti et al., 2009). However, these treatment processes
are not applied in the water treatment processes in Ethiopia.
A study conducted in the rift valley region of Ethiopia detected organochlorine pesticides such as
endosulfan and DDTs soil samples at a concentration of 56 mg/kg and 0.00023 mg/kg,
respectively. Such contamination is known to pose a risk of contamination to the surface and
surrounding water bodies (Westbom et al., 2008). In Ethiopia, the health hazards associated with
pesticide handling are not well understood by both the sprayers and consumers. Misuse of
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pesticides, lack of awareness towards the proper handling of pesticides and poor monitoring
systems and the presence of high amounts of pesticides are the main contributors of water source
contamination (Mekonnen and Agonafir, 2002). Historically, pesticide use by small holder
farmers has been low. Recent developments, however, increased the use of pesticides to diversify
food production and to enhance the international floriculture industry (Amera and Abate, 2008).
In the rift valley regions of Ethiopia, water samples taken from surface waters around
agricultural fields, effluent waters from floriculture and intake of the Zeway water supply system
show that concentrations of pesticides exceeded the European Union drinking water standards
(Jansen and Harmsen, 2011).
To date, there are no comprehensive assessments of pesticide residues from different drinking
water sources upon which consumer risk could be predicted. Therefore, the present study aims at
investigating pesticide residues from different water sources (river, spring, well, and community
taps) that are used for human consumption and to undertake consumers risk assessment.
6.2 Materials and Methods
6.2.1. Study area
One of the sampling areas is the Addis Ababa water supply system. Addis Ababa lies 9°1′48″N
latitude and 38°44′24″E longitude. The city is located at the heart of the country at an
altitude o f 2100 meters above sea level. The city occupies a total area of 540 km2 and has a
complex mix of highland climate regions with annual average temperature of 22.2°C. The
study area encompasses the Legedadi pond which is the main water supply source for Addis
Ababa population. During data collection, geographical positioning system (GPS) coordinates
were recorded on sites where water samples are collected to construct a map of study area for
both Jimma zone and Addis Ababa. The catchment area of Legedadi pond is about 206 square
kilometres, which has similar rainfall and temperature conditions compared to Addis Ababa
figure 6.2.
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Figure 6.2. Map of the study area in Addis Ababa water supply system
The other sampling area is Jimma zone Southwestern Ethiopia water sources (spring, well,
river, and community taps). The study area includes ten districts located in Jimma zone in
which water samples were collected. Jimma zone is located in Oromia region, 1744m above
sea level, at latitude of 7°40’0.01” and longitude of 37°0’0” figure 6.3.
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Figure 6.3. Map of the study areas in Jimma zone, southwestern Ethiopia
6.2.2. Sampling
Water samples were collected from different water sources (springs, river, well) using a grab
sampling method, in which the sampling bottle is submerged at a certain depth in water (15-20
m) and take the required amount of water. Additionally, water samples also taken from treatment
plants before and immediately after treatment including the community taps and distribution
reservoirs. Most of the water sources are surrounded by agricultural fields where chemical
pesticides were applied (personal observation). The water samples were classified in to treated
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(samples taken immediately after treatment, community taps and reservoir) and untreated water
(samples taken from river water, spring and well).
The water samples were also collected from Addis Ababa main water supply source that is the
Legedadi pond and from two small rivers that pass through agricultural fields and join the
Legedadi pond. In the pond just before the water treatment plant three different sampling points
were considered. In the distribution system four sampling sites were considered (one from just
after the treatment, one from central reservoir and two from community taps). A total of nine
sampling points were considered from the source up to the consumption points. The samples
were collected two weeks after the pesticide application. After collection the water samples were
transported using cold box to the laboratory and stored at 4°C until the extraction was done.
6.2.3. Chemicals and reagents
Acetonitrile (99.9%), n-hexane (99%), methanol (100%) and anhydrous sodium sulphate,
saturated sodium chloride and HPLC (high-performance liquid chromatography) water were
used for the extraction and clean-up of the pesticides. Anhydrous sodium sulphate was used to
remove water from the hexane solution and saturated sodium chloride was used to facilitate the
separation of water and hexane solution. Methanol and HPLC water were used to activate the
Sep-Pak cartridges (C18) before extraction of the water samples. Five point calibration curves
were obtained from solutions containing the standard mixture of the pesticide under study (2,4-
D, malathion, diazinon, fenpropimorph and pirimiphos methyl). The standard curves were
prepared using a concentration range of 0.001– 0.1 mg/l.
6.2.4. Extraction and clean-up of the samples
Based on the collected samples, extractions were made in two different ways: liquid- solid and
liquid-liquid extractions. This is important to select a method which gives good recovery for the
pesticides under study.
6.2.4.1. Liquid- solid extraction procedure
Sep-Pak C18 classic cartridge with 360 mg sorbet per cartridge, 55-105 µm particle size and
125A° pore size column was used to extract the samples. The Sep-Pak is silica- based bonded
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phase with strong hydrophobicity which is used to adsorb the analyte of even weak
hydrophobicity from the aqueous solution. For our study, the Sep-Pak was first activated with 1
ml methanol passing through the bonded phase and rinsing with 1 ml HPLC water, sequentially.
Then, one litre of water sample was passed through the cartridge. The Sep-Pak samples were
rolled with Teflon, well labelled and stored at -20°C until the extraction with solvents was done.
The extraction procedures were as follows: the Sep-Pack cartridges, in which a 1000 ml of water
sample passed through, were opened. Then, 10 ml of n-hexane as extraction solvent was pushed
through the Sep-Pak using 10 ml syringe into a flask. Five to six grams of sodium sulphate
(Na2SO4) was added to the flask which is to separate water from the solvent. Then n-hexane was
evaporated to dryness using a rota-vapour at 40°C and reconstituted with 10 ml of acetonitrile to
make the samples amenable for LC-MS/MS injection. Then the extract was put into vials for the
determination of pesticides using LC-MS/MS instrument.
6.2.4.2. Liquid-liquid extraction procedure
In addition to the Sep-Pak samples, raw water samples were collected from each of the sampling
sites. A 250 ml plastic bottle (two bottles for each site) was used to collect raw water samples,
which were rinsed well with distilled water and dried in the air before sample collection. The
bottles were carefully filled with the water samples to overflow without trapping air bubbles in
the bottle. The samples were transported to the laboratory and stored in similar condition with
the Sep-Pak samples.
The liquid-liquid extraction procedure was undertaken as follows: a 500 ml of water sample was
added into a one litre separator funnel and 100 ml of n-hexane was added followed by 10 ml of
saturated sodium chloride (NaCl). Samples were agitated by hand to enhance the contact of the
solvent with the pesticides. After 10 minutes, the hexane and water medium were separated and
the water phase was removed at the bottom of the separator funnel. The hexane solution passed
through 15 grams of anhydrous sodium sulphate into a 250 ml flask in order to absorb any
remaining water from the hexane solution. For the completeness of the extraction, this procedure
was repeated once; again the hexane solution was diluted up to 250 ml n-hexane and then
evaporated to dryness using rota-vapour at 40°C and reconstituted with 10 ml of acetonitrile. The
extract was then transferred to LC-MS/MS vials for analysis.
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6.2.5. Analytical equipment and chromatographic separation
For the determination of the pesticides under study, liquid chromatography with double mass
spectrometer (LC-MS/MS), with water acuity UPLC instrument was used. The respective
chromatographic conditions were as follows: The column was water HSS T3 (1.8µm) and an
injection volume of 5µl. The oven temperature was 40°C and a flow rate of 0.4 ml min-1. The
detector of triple quadruple mass spectrometer with Electrons Spray Ionization (ESI), and with a
potential of 500 volt and a temperature of 500°C. The scan type was multiple reactions
monitoring mode (MRM). The respective ESI, Ions (parent and daughter ions) of the pesticides
in this study were indicated in table 6.1.
Table 6.1. The parent and daughter ions for each pesticide analysed in water samples using LC-
MS/MS
Parent ions Daughter ion ESI Cone voltage Collision energy
Pesticide
m/z m/z charge (eV) (eV)
2,4-D 218.9/220.9 160.7/162.7 Negative 20/20 11/11
diazinon 305 95/169 Negative 31/31 35/22
malathion 331 99/127 Positive 20/20 24/12
fenpropimorph 304.2 57/147 Positive 50/50 30/28
pirimiphos methyl 306 100.1/164.1 Positive 30/30 32/22
m/z = mass to charge, eV = electron volt, ESI = Electron Spray Ionization
During the analysis, analyte concentration below the limit of detection (LOD), but giving signals
were reported as less than the LOD. Moreover, the observed concentrations from the instrument
were compared with the norm value of 0.1 μg/l set for single pesticides by the European
Commission (EU) and World Health Organization (WHO) guidelines (World Health
Organization,2008, World Health Organization, 2011, Northern Ireland Environment Agency,
2011).
6.2.6. Screening
Screening was carried out before the actual water samples were analysed to determine which
pesticide was presented in the samples. During the screening, pesticides such as DDT,
deltamethrin, permethrin, cypermethrin were also included, while only fenpropimorph, diazinon,
malathion and 2,4-D were detected. The pesticides that are not detected may be due to their low
water solubility. The screening was performed by mixing a small portion of sample from each
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site and passing it through the Sep-Pak cartridge. Subsequently, the quantitative determination
and identifications of each pesticide was worked out for all the samples using peak area and
retention time, respectively.
6.2.7. Quality control
Representative samples were collected in clean materials for the extraction and analysis. Sep-Pak
C18-column extraction was made within 48 hours of sample collection, which helped to avoid
degradation of the pesticides and to determine the accurate quantity of the residues. Both the
Sep-Pak cartridges and the raw water samples were transported in a cold box with ice pack to the
laboratory and were stored at refrigerator temperature until the extraction was made. The glass-
ware materials used were kept clean to avoid cross contamination and all the reagents used were
analytical grade.
Before the analysis of the samples, the method validation was undertaken by determining the
recovery of pesticides, limit of detection and quantification and the regression coefficient. To
determine the accuracy of the method, recovery studies were performed by spiking a known
amount of the standard mixture of pesticides in ultra-pure water. The percent recovery was
calculated by dividing the recovered concentration by the spiked concentration. The limit of
detection (LOD) and the limit of quantification (LOQ) were also determined by taking the signal
to noise ratio (S/N) into account. For LOD the lowest concentration when S/N ratio is over 3 was
considered, while for LOQ the lowest concentration when S/N ratio is over 10, was considered.
6.2.8. Statistical analysis
Statistical analysis was performed using SPSS (Version 20). Before the actual data analyses,
normality of the data was checked using histogram. The concentration of all the four pesticides
(2,4-D, malathion, diazinon and fenpropimorph) showed skewness. As the result, a non-
parametric statistical analysis was followed. Kruskal-Wallis test was applied to compare the
distribution of pesticides residues among the water sources (spring, well, river, pond, and
community taps). The level of significance was set at p-value = 0.05. The results were presented
using a box and whisker plot.
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6.2.9. Risk assessment
To understand the intake of pesticides under study from consumption of water, risk (both chronic
and acute) was assessed for human population with different groups (Adult, Children and
Infants).
6.2.9.1. Chronic risk assessment
The estimated daily intake (EDI) was calculated from the residue of pesticides in water and the
consumption of water. Then to characterize the risk EDI was compared with acceptable daily
intake (ADI) for each pesticide. The ADI is an estimate of the daily maximum intake of a
substance over a lifetime that will not result in adverse effects at any stage in human life span. It
is expressed on a body weight bases (Food and Agriculture Organization of the United Nations
and World Health Organization, 2009).
The EDI (mg/kg bw/day) is found by multiplying the residual pesticide concentration in drinking
water (µg/l) with the daily intake of water and dividing it with the average body weight of
consumers based on the equation below (Mahmood et al., 2014). The average daily intake of
drinking water for a 60 kg adult, 10 kg child and 5 kg infant were 2 l, 1 l and 0.75 l per day,
respectively (Younes and Galal-Gorchev, 2000).
EDI is calculated by the following formula.
EDI = Cw * Iw/Bw
Where:
Cw = concentration of the pesticide in water (µg/l)

Iw = daily intake of water (l/day)
Bw = Body weight (kg)
For our study, the maximum concentration in water was considered (worst case scenario) to
calculate the EDI. From our findings, EDI greater than ADI indicates a potential chronic human
health risk for consumers as reported by (Darko and Akoto, 2008).
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6.2.9.2. Acute risk assessment
The short term exposure to pesticide residues was assessed via oral intake of drinking water
containing the pesticides under study. The acute reference dose (ARfD) is an estimate of an oral
exposure of a chemical for short term duration (usually 24 hours). The ARfD is used as a
measure of acute toxicity of pesticides. The presence of pesticides in water sources may not be
uniform or under average expected level. The non-uniform distribution of residues within water
may result in some individuals being exposed to above average levels for short periods of time
resulting in an acute risk (Renwick, 2002). The EDI for short term exposure of the pesticides
under study was calculated by the same equation of the chronic exposure and compared with
ARfD. EDI greater than ARfD is considered as, there is a potential acute human health risk for
consumers.
6.3 Results
6.3.1 Method validation results
The method validation results are given in table 6.2. The regression coefficient (r2) was greater
than 0.995 for all the pesticides under study which indicates that there is a perfect linearity of the
standard curves. The mean percent recovery (% recovery) was in the range of 72.2-92.0%. The
method validation results met the requirements of European commission (EU)
(SANCO/12571/2013, 2013). This revealed that the method was accurate to determine the
pesticides under study. The LOD and LOQ were in the range of 0.003 to 0.0150 and 0.01 to
0.050 µg/l, respectively. This indicates that the method is able to detect up to a minimum
concentration of 0.0003µg/l.
Table 6.2. Method validation results

Pesticides r2 % recovery LOD (µg/l) LOQ (µg/l)
2,4-D 0.9998 85.400 0.0030 0.010
diazinon 0.9995 83.000 0.0003 0.001
malathion 0.9997 92.000 0.0003 0.001
pirimiphos methyl 0.9999 82.000 0.0150 0.050
fenpropimorph 0.9998 77.200 0.0003 0.001
LOD = limit of detection, LOQ = limit of quantification
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6.3.2 Pesticide residue in drinking water sources in Jimma zone, southwest

Ethiopia
Water samples collected from all water sources were analysed for the presence of pesticides.
From the results, 2,4-D, diazinon, malathion, fenpropimorph and pirimiphos methyl were
detected. From these pesticides pirimiphos methyl was detected in only two water sources such
as well and spring water with a concentrations of 0.33 and 168.20 µg/L, respectively. A
summary of the residues of the pesticide in the water samples is given in table 6.3. The average
concentrations of 2,4-D ranged from 2.27 to 2.94 µg/l among the different water sources, where
the highest concentration was found in well water. The highest concentration of diazinon (~5.65
µg/L) was registered in the river water sample of which Jimma town receives water for its
municipal water treatment.
Table 6.3. Summary statistics for the pesticide residue (µg/l) in drinking water sources of Jimma
zone
Pesticides Water source Min Max Mean StDv
River water 1.590 3.971 2.666 0.796
Spring 1.574 5.703 2.690 0.931
2,4-D Tap water 1.412 3.472 2.270 0.517
Well water 1.559 5.797 2.939 1.361
Distribution reservoir 1.368 3.365 2.354 0.583
River water 3.000 7.477 5.646 1.321
Spring 0.0001 15.633 1.592 3.229
diazinon Tap water 0.0001 9.644 2.023 2.187
Well water 0.0001 11.491 2.197 3.863
River water 1.854 9.042 5.028 1.940
Spring 0.008 7.036 1.711 1.622
fenpropimorph Tap water 0.008 7.773 2.681 2.279
Well water 0.008 11.491 2.408 3.290
River water 0.0001 50.162 7.658 11.945
Spring 0.0001 67.439 7.252 12.707
malathion Tap water 5.1000 11.948 7.713 2.745
Well water 0.0001 105.031 13.947 27.839
StDv = Standard deviation, river water = intake for treatment
The mean concentration of malathion ranged from 7.25 to 13.95 µg/l, in which the highest
concentration was observed in the well water. Fenpropimorph was mainly observed in the river
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water. The detection of the pesticides at different concentration indicates the contamination of
the water environment and which will result in exposure of the communities who consume water
from these sources. These pesticides may come to the water bodies from the surrounding
agricultural fields. Therefore, building up of buffer zones and strict control of the pesticides
applied near to the water sources is important.
Kruskal-Wallis test indicates that the distribution of diazinon and fenpropimorph among the
water sources were significantly different (p-value < 0.004) (figure 6.4). But other pesticides had
no statistical difference between the water sources. On the other side, water samples taken from
the Gilgel Gibe River before treatment, contaminated by high concentrations of diazinon and
fenpropimorph compared to the well and spring water sources. This indicates that the
environment is highly contaminated by fenpropimorph and diazinon or these two chemicals may
be applied more in the surrounding agricultural areas.
River Spring Tap Well Reservoir River Spring Tap Well Reservoir
Water sources Water sources
Figure 6.4.Concentration distribution of the pesticides under study (µg/L) among different water
sources. The inner box indicates the mean, the outer box indicates mean ± standard error and the
whiskers indicate the 95% confidence interval
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As it can be depicted from the box and whisker plot, 2,4-D and malathion residues were much
higher in the well water compared to other pesticides under study. This may be due to, long
residence time of the water in well.
6.3.3 Exposure assessment of population in Jimma zone to pesticide residues in

water
Human exposure was assessed based on the residue of each pesticide in the water sources and the
average water consumption table 6.4. From the findings, the estimated daily intake (EDI) for
adults, children and infants were below the ARfD for all the pesticides under study. The EDI for
diazinon was above ADI for adults, children and infants, while all other pesticides under study
were below the ADI. The ADI and ARfD for each pesticides were taken from (FOOTPRINT-
IUPAC-PPDB, 2015).
Table 6.4. The estimated daily intake of pesticides by population Jimma zone from consumption
of water
EDI (µg/kg bw/day) ADI ARFD
Water Max. Conc.
Pesticides (µg/kg (µg/kg
sources (µg/l) Adult Children Infants
bw/day bw/day
River water 9.042 0.301 0.904 1.356
Spring 7.036 0.235 0.704 1.055
fenpropimorph 3 30
Tap water 7.773 0.259 0.777 1.166
Well water 11.491 0.383 1.149 1.724
River water 3.971 0.132 0.397 0.596
Spring 5.703 0.190 0.570 0.856
2,4-D 50 750
Tap water 3.472 0.116 0.347 0.521
Well water 5.797 0.193 0.579 0.869
River water 7.477 0.249 0.748 1.122
Spring 15.633 0.521 1.563 2.345
diazinon 0.2 25
Tap water 9.644 0.321 0.964 1.447
Well water 11.491 0.383 1.149 1.724
River water 50.162 1.672 5.016 7.524
Spring 67.439 2.248 6.744 10.116 20 300
malathion
Tap water 11.948 0.398 1.195 1.792
Well water 105.031 3.501 10.503 15.755
EDI = Estimated Daily Intake, ADI = Acceptable Daily Intake, ARfD = Acute Reference Dose,
Values in bold indicate EDI above health based guidance values.
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6.3.4 Pesticide residues in Addis Ababa water sources
The findings of the present study showed the detection of 2,4-D, diazinon and fenpropimorph
consistently from the source (river, pond) up to the distribution points in the Addis Ababa water
supply system table 6.5. The maximum concentration of fenpropimorph (164 µg/l) and 2, 4-D
(127 µg/l), were observed in samples collected from the two small rivers while the maximum
concentration of diazinon (96.23 µg/l) was observed in Legedadi pond. Regarding the
community taps 2,4-D, diazinon and fenpropimorph were detected at concentration of 4.728 μg/l,
0.132 μg/l and 0.126 μg/l, respectively. The conventional treatment process decreases some
amount of the pesticides however, complete removal was not observed. As a result the pesticides
still detected up to point of consumption (community taps) which may cause a health problem
for human health.
Table 6.5. Concentration (µg/l) of 2,4-D, diazinon and fenpropimorph for each sampling point
Pesticides Water sources Min Max Mean StDv
2,4-D River 11.200 127.000 69.100 81.800
Pond 7.040 30.200 14.900 13.200
Distribution 4.800 6.590 5.700 1.250
Reservoirs 5.810 6.120 5.970 0.200
Community taps 0.230 4.730 0.597 1.260
diazinon River ND ND ND ND
Pond 0.014 96.230 32.100 55.500
Distribution 0.065 11.000 5.500 7.770
Reservoirs 0.250 1.240 0.633 0.800
Community taps 0.034 0.132 0.083 0.600
fenpropimorph River 112.000 164.000 138.000 36.760
Pond 0.430 76.900 26.000 44.000
Distribution 0.173 0.335 0.254 1.100
Reservoirs 0.107 0.110 0.109 0.020
Community taps 0.105 0.126 0.116 0.140
ND = not detected, StDv = Standard Deviation
6.3.5 Exposure assessment of population in Addis Ababa to pesticide residues in

water
The human exposure for adults, children and infants in Addis Ababa is expressed as the EDI and
is presented in table 6.6. Adults, children, and infants consuming water from the two small rivers
and children and infants consumed water from the Legedadi pond, were exposed above the
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health based guidance value (EDI > ADI), but less than the ARfD. The EDI for 2,4-D from
source up to the community taps were below the ADI and ARfD. Regarding diazinon, the EDI
was above the ADI but below the ARfD for adults; children and infants consuming water from
the pond. For the exposure assessment the results of the two rivers were combined assuming that
the farmers applied similar pesticides as they produced similar crops (personal observation). This
hypothesis was confirmed by the obtained study results. The EDI above the health based
reference values indicates that there will be health problems for people drinking water from these
sources.
Table 6.6. The estimated daily intake of the detected pesticides in water sources
Max. EDI (µg/kg bw/day ADI ARfD
Pesticides Water sources Conc. (µg/kg (µg/kg
Adult Children Infants
(µg/l) bw/day bw/day
River 164 5.467 16.400 24.600
fenpropimorph Pond 76.868 2.562 7.687 11.530 3 30
Community Taps 0.126 0.004 0.013 0.019
River 126.597 4.220 12.660 18.990
2,4-D Pond 30.247 1.008 3.025 4.537 50 750
Community Taps 4.728 0.158 0.473 0.709
River * * * *
diazinon
Pond 96.198 3.207 9.620 14.430 0.2 25
Community Taps 0.132 0.004 0.013 0.020
* = exposure was not calculated for non- detected, NA = Not Applicable, ADI = acceptable daily
intake, ARfD = acute reference dose, values in bold indicate EDI > ADI
6.4 Discussion
The result of the present study confirmed the presence of 2,4-D, malathion, diazinon, pirimiphos
methyl and fenpropimorph, in drinking water sources from Jimma zone southwestern Ethiopia.
The herbicide 2,4-D was detected in all water samples and this may be due to the time of sample
collection (July-August), in which weed control pesticides are applied in agriculture. Higher
concentration of 2,4-D and malathion were detected in well water, while diazinon and
fenpropimorph occurred more in river water. This may be due to the rainy season in which
erosion may bring pesticides to the water sources from the surrounding agricultural fields.
The results from Addis Ababa drinking water sources (rivers, pond, reservoir and community
taps) indicate the presence of 2,4-D, diazinon, and fenpropimorph from source up to the
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community taps. The highest concentration of 2,4-D and fenpropimorph were found in the river
water passing through the agricultural fields. This may be attributed to the high amount of
rainfall that occurred during the months preceding pesticide application and the lack of buffer
zones around the Legedadi pond, which may help to prevent the contamination of the water
body. The herbicide 2,4-D is used mainly for the control of dicotyle weeds in crops such as teff
and wheat, which are mostly grown around the studied water basin (Kippie Kanshie, 2002).
In addition to this, the presence 2,4-D is explained by its high water solubility (900 mg/l, at
25°C), its low potential to move from the water into the air. Concerning 2,4-D, the result of this
study was not consistent with earlier results obtained in Canada (Donald et al., 2007). This
discrepancy may be attributed to the difference in the size of the water body, the pesticide
handling systems, the scale of application, the climatic conditions and the amount and duration
of the application.
Diazinon and malathion are important organophosphate insecticides in agricultural crops and
mainly applied for the control of cockroaches, ants and fleas. Their presence in the water bodies
from Jimma and Addis Ababa may be explained by transport from the point of application by
drifting to the water and its stability to photolysis. According to (Fadaei et al., 2012), degradation
of organophosphate pesticides by photons from the sun is very low in natural water. From a
study done in Iran, diazinon and malathion were also detected in drinking water sources
(Shayegehi et al., 2007). Chloripyrifos, malathion, alachlor, dimethoate and metribuzin were
detected at varying concentration in water samples collected from India (Sinha, 2011). The
residue of diazinon in the present study was lower than what is detected in surface water of Iran
(Fadaei et al., 2012) and Venezuela (Flores-García et al., 2011). However, the concentration was
higher than what is observed in Lebanon (Kouzayha et al., 2013).
Fenpropimorph is a fungicide which is mainly used for the control of fungal diseases like rust in
cereals and in barley leaf. Its presence in drinking water may be explained due to leakage from
obsolete pesticide stocks, biological desorption of fenpropimorph residues and its stability in the
water. The residue of fenpropimorph was present in all Jimma water sources and in rivers and
ponds in Addis Ababa water sources. The residues from Addis Ababa were considerably higher
than the results obtained in Demark drinking water sources (Schriever et al., 2007). This may be
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attributed to the lack of awareness of Ethiopian farmers to follow good agricultural practices and
to handle pesticide in addition to application conditions and local geological, hydrological and
meteorological conditions.
The mean concentration of the pesticides from our study areas exceeded the European drinking
water standard of 0.1 μg/l for individual pesticides and 0.5 μg/l for the sum of all the detected
pesticides except for the mean concentration of diazinon in rivers. Based on WHO guidelines,
the maximum allowable concentration for 2,4-D, diazinon and fenpropimorph in the drinking
water is 30 μg/l, 1 μg/l and 0.1 μg/l, respectively (World Health Organization, 2008). The
concentrations of 2,4-D and diazinon from Jimma water sources appear to be below the WHO
standard, while diazinon and fenpropimorph in the Addis Ababa water sources were above the
guide line. The results above the guidelines indicate that there is a high contamination of the
water sources by the pesticides under study. The contamination may come from different
agricultural fields surrounding the water sources and by poor application and storage of
pesticides by users.
From the results in the Jimma zone, high concentrations of the pesticides were detected in
untreated water sources (well and river water). This may be attributed to the fact that, the
residence time for water is long in the well, which limites the pollutant flow. While for river
water the eroded soil from the surrounding agricultural areas may directly enter into the water
sources. Regarding the Addis Ababa water sources, there is a continuous reduction of the
residues of each pesticide from the source (Legedadi pond and rivers) up to the community taps.
This may be due to the longer distance of Addis Ababa from the agricultural area and
degradation of the pesticides may happen in between. Additionally, the results may be attributed
to the larger volume of water in the pond and the deposition of the pesticides with the sediment.
The treatment facilities in the Legedadi treatment plant, may also contribute to further reduction
of the pesticide residue in the water.
The detection of these chemical pesticides in the water sources may affect the health of
consumers in Ethiopia who uses these water sources for drinking. From the results in the present
study, the risk estimation of the pesticides in water from Jimma zone indicates that the EDIs for
2,4-D, malathion and fenpropimorph, were below the ADI and ARfD for all population groups.
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This means that there is no acute and chronic health problems expected for consumers. However,
the EDI of diazinon for adults, children and infants consuming water from the river, well, spring
and even in community taps, were above the ADI, but below the ARfD. This indicates that there
is a chronic health problem for the population in Jimma zone. A similar chronic risk of diazinon
and malathion from drinking water is also reported for the Iranian population (Shayegehi et al.,
2007). Organophosphate pesticides such as diazinon, malathion and others are known as
neurotoxic substances inhibiting acetylcholine esterase enzyme and they have genotoxic effects
causing DNA damage and also have cytotoxic effects on human health (Muranli et al., 2015).
From the results of Addis Ababa water sources, the EDIs of fenpropimorph for adults, children
and infants consuming water from the rivers and for children and infants consuming water from
the Legedadi pond were above the ADI, but below the ARfD. The EDI for 2, 4-D for the
population who consume water from sources up to the community taps were below the ADI and
ARfD. Regarding diazinon, the EDI was above ADI but less than the ARfD for adults, children
and infants consuming water from the pond. As is described, the EDI for all the pesticides under
study were below the ARfD. This indicates that there is no short term risk for consumers.
However, this does not grantee safety for the chronic exposure of these pesticides as confirmed
by the results of our study.
Due to cumulative effects of pesticides, adverse human health effects may occur through chronic
exposure. This raises the need for a greater attention to create awareness about the use of
pesticide by farmers across the water resources. Care should be taken to decrease the risk of
these pesticides by establishment of buffer zones (15 to 30 m around the water sources) which
minimize pesticide reliance to water sources. Moreover, maintain the application equipment in
good working condition and avoidance of poorly timed application of pesticides is important for
decreasing contamination of water as is explained by Waskom, (2010).
6.5 Conclusions
The result of this survey depicts that the drinking water sources are contaminated with diazinon,
2,4-D, fenpropimorph and malathion from the source up to the community taps. Some of the
detected concentrations of pesticides exceed the European drinking water standard. This
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indicates that there is intensive use of these pesticides around the study area. The rural
communities in Ethiopia, who are using untreated water sources for drinking (well, spring, rivers
and ponds), may suffer more than urban communities from health risks. The Jimma and
Legedadi water treatment systems reduce pesticides in the water however, complete removal was
not observed. This indicates that the conventional water treatment is less effective for the
removal of pesticides from water. Based on WHO and EU drinking water guidelines, high
amounts of residues still remain in the distribution systems, reservoirs and community taps.
Concerning human health risk estimation, there is no acute risk for people drinking water from
all water sources as the estimated daily intake is less than the acute reference dose (EDI <
ARfD). However, long term risks were observed especially for diazinon and fenpropimorph for
which, the EDI > ADI for the Jimma and Addis Ababa population. A comprehensive monitoring
and source oriented remediation action is required to reduce the level of pesticide residues and to
minimize in particular the long term health risks on human health.
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Chapter Seven: Exposure to DDT from khat (Catha edulis) consumption: a double burden to
consumers in southwestern Ethiopia
Chapter seven: Exposure to DDT from khat (Catha edulis) consumption: a

double burden to consumers in southwestern Ethiopia
Redrafted from: Mekonen S., Ambelu A., Negassa B. and Spanoghe P. Exposure to DDT and Its
metabolites from Khat (Catha edulis) consumption: a double burden to consumers in
Southwestern Ethiopia
Submitted to: PLOS ONE and it is under review
113
114
Abstract
Khat (Catha edulis) is a highly consumed crop for countries located in the horn of Africa.
Naturally, khat is known to have different adverse health effects on regular consumers. On top of
that khat used for human consumption may contain pesticide residues which may also impose
adverse health effects. The present study aimed to investigate DDT and its metabolite (p,p’-
DDE, p,p’-DDD, o,p’-DDT and p,p’-DDT) residues in khat and human exposure of khat
consumers. A khat consumption survey was undertaken using the 24 hour recall method. Khat
leaves with tender stems samples were collected from the local market located in different
districts of the Jimma zone in southwestern Ethiopia. Human exposure to this pesticide was
determined using the probabilistic exposure assessment method. From the results, eighty percent
(80%) of the khat samples contained total DDT. Some of the residues were above the maximum
residue limit (MRL) set by Codex Alimentarius and most of the samples contained DDT residues
above the European Commission MRL set by European Food Safety Authority (EFSA). The
concentration of p,p’-DDE and p,p’-DDT in khat were in the range of 0.033 to 0.113 and 0.010
to 0.026 mg/kg, respectively. The metabolite (p,p’-DDE) was detected at high concentration
compared to the parent compound (p,p’-DDT). This indicates that there was historical use of
DDT in the study area. From probabilistic exposure analysis, the mean and the 97.5 percentile
(p97.5), estimated daily intakes (EDIs) of total DDT were 0.002 and 0.006 mg/kg bw/day, which
were below the provisional tolerable daily intake (PTDI). However, this does not guarantee full
safety, as there may be contamination of other commonly consumed food items resulting in a
cumulative DDT exposure. Khat leaves used for chewing had considerable concentrations of
DDT and its metabolites which impose an additional health risk. Stewardship and education of
farmers growing khat and continuous monitoring of pesticides including DDT in khat and other
commonly used agricultural produce is important to minimize particularly the chronic health risk
to consumers.
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7.1 Introduction
The use of pesticides is considered to be a vital practice since they increase crop outputs,
improve the quality of products, and decrease the incidence of illnesses propagated by insects
such as malaria and typhus (Beceiro-González et al., 2012). However, the residue of the applied
pesticides remains in the environment. The contamination of food items by hazardous
substances, in particular the presence of residues of persistent organochlorine pollutants (POPs)
is a worldwide public health concern (Kathpal and Kumari, 2009). Since certain pesticides are
hazardous and toxic to human health, any residue remaining in or on food can pose danger to
humans and may cause side effects (Aktar et al., 2009). Despite the ban of DDT since the 1970s,
the use of this chemical has continued in certain parts of the world and in particular, in
developing countries for the control of malaria due to its effectiveness and cheap price.
In the past decades, the intensive use of DDT for agricultural and anti-malarial purposes in
developed and developing countries has resulted in significant contamination of food products
(Nakata et al., 2002). The residue of DDT in different foodstuffs has been investigated in various
countries to understand the status of contamination and to evaluate the possible impact on human
beings. Due to its persistence and bioaccumulation properties, DDT can be transferred and
magnified to higher trophic levels through the food chain (Man et al., 2013).
Because of their persistent nature, the residues of DDT and its metabolites still appear as a
contaminant in both plant and animal based foods such as medicinal plants (Rodrigues et al.,
2005), tea leaves (Amirahmadi et al., 2013), tobacco (Cai et al., 2005; Ghosh et al., 2014;
Rahman et al., 2012), grapes (Turgut et al., 2011), wheat (Daba et al., 2011), and maize
(Mekonen et al., 2015). Additionally, DDT can be detected in fruits and vegetables such as
papaya, banana, mango, pineapple, tomato, lettuce, cabbage, green pepper, onion, cucumber
(Bempah et al., 2012). Moreover, from a study done in Tunisia and Ethiopia, DDT and its
metabolites are detected in cow and human milk (Ennaceur et al., 2007; Gebremichael et al.,
2013).
Exposure to pesticide residues from food consumption is assumed to be five orders of magnitude
higher compared to exposure through air or drinking water (Claeys et al., 2011). Pesticide
116
residues in foods have received great attention as one of the most important food safety issues
considered for consumers (Zhang et al., 2011). Pesticide residues can affect consumers more
specifically, when food items are freshly consumed without any treatments or preparation
processes which may reduce the pesticide residues (Daba et al., 2011). Khat chewers consume
the fresh leaf without any processing.
Khat (Catha edulis), is a flowering evergreen tree or large shrub of the Celastracea family. It is a
well-known natural stimulant and is chewed as a refreshment, excitement and euphoria. Even
though, khat is consumed for different purposes, there is no data on the exact number of Khat
chewers on a worldwide scale. However; the number of consumers is increasing in time (Dessie,
2013). Khat is known to have cathinone, cathine and norephedrine in which cathinone is the
principal psychoactive component. For this reason khat is blamed for different adverse health
effects (Damena et al., 2011; Douglas et al., 2011; Dessie, 2013). This is a reason that khat is
illegal in most European and North American countries (Armstrong, 2008). The health effects
due to the natural content of khat is a primary burden to regular khat users in the worldwide,
especially in countries located in the horn of Africa, such as Ethiopia, Somalia, Kenya, Eritrea,
Djibouti and Uganda, as well as across the Arabian Sea such as Yemen and Saudi Arabia (Al-
Mugahed, 2008).
Additionally, khat leaf is expected to contain higher pesticide residues compared to other
processed food groups of plant origin since it is consumed raw without any processing steps like
washing or cooking. High residues of DDT were reported in khat leaves collected from specific
farm lands in Ethiopia (Daba et al., 2011; Ligani and Hussen, 2014). According to El-Zaemey
and his colleagues (El-Zaemey et al., 2015), Ethiopian khat cultivators use DDT and other
pesticides as khat growing plant protection products. Despite the problems related to khat use,
only a few studies are published which present very high concentrations of p,p’-DDT, ranging
from 999.0 to 141.2 µg/kg in khat samples collected in the eastern part of Ethiopia (Daba et al.,
2011). Another study done in the southern part of Ethiopia also indicates up to 44.8 µg/kg of
p,p’-DDT in khat samples (Ligani and Hussen, 2014). Jimma zone is one of the khat growing
areas in the southwestern part of Ethiopia where khat chewing is a deep-rooted tradition in the
117
population (Damena et al., 2011). Therefore, the main aim of the present study is to determine
the exposure of khat chewers to DDT and its metabolites.
7.2 Materials and Methods
7.2.1 Study area
The present study was conducted in Jimma zone, Southwestern Ethiopia. Jimma zone is one of
the khat growing areas and khat is commonly intercropped with other agricultural crops such as
maize and teff (Dessie, 2013). The study area includes six districts of Jimma which are
considered as a source of Khat for local markets figure 7.1. Jimma town was selected to conduct
the khat consumption survey. The town is located at 350 km to the southwestern part of the
capital Addis Ababa. It is found on approximately 7°41′ N latitude, 36° 50′ E longitude and an
average altitude of 1,780 meters above sea level. According to 2014-2015 report of
Central Statistical Agency (CSA), the projected total population of the town is 170,955
(Male = 85,695 and Female = 85,260) with 85,260 as the number of households.
Figure 7.1 Map of the study area in Jimma zone, Southwestern Ethiopia
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7.2.2 Sample size determination for consumption survey
The source population included all the people living in Jimma town. The study population were
khat chewers that are selected from source population. Total sample sizes of 423 khat chewers
were selected using a single population proportion formula. To maximize the sample size,
prevalence of khat chewers was considered as 50%. Ninety five percent certainty (95%)
confidence interval and 5% margin of error was taken. Ten percent non-response rate was added
to the sample size as a contingency.
n=
Where:
Z = coefficient for 95% level of confidence, Z = 1.96

P = proportion of the population = 50%, because similar studies were difficult to find and
it was assumed that 50% of khat chewers were exposed to DDT and its metabolites
residue,
E = margin of error (5%) = 0.05.
n = 384.16,
10% of non-response rate was then added to get the final sample size (nf).
Hence, nf = 384.16 + (384.16X0.1) = 384.16 + 38.416 = 423
7.2.3 Consumption survey
After determination of the sample size, a consumption survey was undertaken to assess the daily
intake of khat by 423 consumers. The survey was done in randomly selected households. Socio-
demographic (age, sex, educational status, marital status and body weight) data of the chewers
were collected during this household survey. The khat consumption data were collected using a
repeated 24 hour recall method. An interview administered by trained interviewers using a face-
to-face interview was undertaken for two non-consecutive days separated by fifteen days. The
amount of chewable khat leaves with tender stems consumed per day was calculated. The
119
average of both recall days was used to estimate khat consumption on a daily basis. The mean
daily consumption (g/kg bw/day) for each khat chewer was calculated by dividing the average
daily consumption of khat (g/day) by his/her body weight (kg). The body weight of the khat
chewers was obtained by interviewing the study participants.
7.2.4 Data handling and analysis
Data analysis was performed using the Statistical Package for Social Sciences (SPSS for
Windows version 20). The significant level was set at p-value < 0.05. Exposure analysis of the
khat chewers was conducted using probabilistic exposure analysis method; in the @risk
statistical software for Microsoft excels 2010 and Monte Carlo simulation model.
7.2.5 Khat sampling
Khat leaves with tender stem samples were collected from local markets in Jimma zone
southwestern Ethiopia by interviewing the sellers regarding the source of the khat to the market.
Chewable parts of khat samples (50-100 g each) were packed in polyethylene plastic bags and
labelled accordingly to indicate the market source and transport the samples to the laboratory.
Then the samples were dried under shade, grinded using mortar-pestle and stored at 4 °C until
analysis.
7.2.6 Reagents and materials
Analytical grade acetone (99.5%), ethyl acetate (99.5%) and n-hexane (95%) were obtained from
BDH limited Poole (BDH AnalaR®)), and were used as solvent for extraction of the khat
samples. Sodium chloride (NaCl) and anhydrous sodium sulphate (Na2SO4) were used for
removal of water from the sample. The analytical method for the extraction of the khat samples
was Solid Phase Extraction (SPE) with column chromatography clean-up, for the removal of
organic acids and polar pigments among other compounds. Standards of DDT metabolites of the
highest analytical purity (p,p’-DDE (99.9%), p,p’-DDD (99.3%), o,p’-DDT (100%), and p,p’-
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DDT (99%), were obtained from Supelco and delivered by Sigma-Aldrich Logistics and used for
the preparation of the calibration curves.
7.2.7 Quality control
For the analysis of the khat samples, we adopted the method for pesticide residue analysis in khat
from Daba et al. (2011), with slight modification. The quantitative determination of the pesticide
residue in khat was done based on the external standard method. The chemicals and reagents
were pure and analytical grade. The calibration curves were obtained by injecting five different
concentrations of the pesticide standards in a range of 0.004-0.08 mg/l. The regression
coefficient (r2) was > 0.995 for all DDT metabolites. Identification and quantification of the
pesticides were done based on the retention time and peak area, respectively.
7.2.8 Extraction and clean-up of khat samples
The modification of the method from Daba et al. (2011) were; 1) at the end of the extraction
procedure we used 2 ml n-hexane for solvent exchange to make the samples amenable for GC-
ECD injection. 2) We used the highly sensitive instrument (GC-ECD) for the determination of
organochlorine pesticides like DDT as explained by (Oliveira et al., 2012) instead of GC-MS.
The analytical procedure is explained in figure 7.2.
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Figure 7.2. Flow-diagram for the sample preparation, extraction, clean-up and analysis of khat
samples
7.2.9 Analytical equipment
Total DDT and its metabolites were determined by capillary gas–liquid chromatography with
electron capture detector (GC–ECD; Agilent Technologies 6890N) in similar chromatographic
conditions with the previous study worked out by Mekonen et al. (2015). After the analysis, the
concentration of total DDT was determined by summing up its metabolites (p-DDT, p,p’-DDE,
p,p’-DDD and o,p’-DDT).
7.2.10 Exposure assessment of khat consumers
To evaluate the safety of khat consumers regarding pesticide residues, the exposure was assessed
using the probabilistic exposure analysis method. The exposure assessment in the present study
was done for the real khat consumers. Total DDT was not detected in 20% of the khat samples.
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According to Medeiros Vinci et al. (2012), when there are non-detected (ND) pesticide residues
in food items, dietary exposure has to be done according to three scenarios such as the upper
bound considers non-detected sample values which equal the limit of detection (ND = LOD),
medium bound for which non-detected values equal half of the limit of detection (ND = 1/2
LOD) and lower bound for which non-detected values equal zero (ND = zero). As there was no
significant difference in the lower, medium and upper bound scenarios in the exposure analysis
(p-value < 0.05), the results of the probabilistic exposure analysis were only done for the upper
bound (worst case) scenarios.
7.2.11 Probabilistic exposure analysis
The exposure of consumers to a number of pesticides was evaluated in more detail by the
probabilistic risk assessment approach. This technique involves random sampling of each
probability distribution within the model to produce hundreds or even thousands of scenarios
(iterations or trials) (Claeys et al., 2011). A probabilistic exposure analysis was conducted using
@Risk® 5.5 software program for Microsoft Excel 2010 (Palisade Corporation, USA). As
mentioned in chapter four, the consumption and residue distributions were combined to give an
exposure distribution and Monte-Carlo simulation ran with 100,000 iterations.
The results from the probabilistic exposure analysis were compared with the provisional
tolerable daily intake (PTDI = 0.01 mg/kg bw/day) of DDT for every agricultural crop set by
Codex Alimentarius (FAO/WHO, 2013). The mean and p97.5 values were considered as the
average and high consumer exposure. The results above the health based reference value were
taken as an indication for human health risk.
7.3 Results
7.3.1 Socio demographic results of the khat consumers
A total of 423 (male = 70%) and female = 30%)) khat chewers were included in the study. All
study participants responded to the questioners, giving a response rate of 100%. From the Mann-
Whitney U test, there is no significant difference in consumption of khat between male and
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female chewers (p-value = 0.141). According to this study, 64.4% of the chewers were in the
age group between 20-39 years and 53.0%, 33.3%, 9.5% and 4.2% were married, single,
widowed and divorced, respectively. Concerning the educational status, out of the total
respondents 84.7% had a higher grade completed (above elementary school), 5.4% of the
participants were illiterate, and 9.9% were able to read and write. This indicated that most of the
khat consumers were in the young age groups, married and educated.
According to the present study, 72% of khat consumers spent more than fifty Ethiopian birr per
day to buy khat and 26.2% and 24.5% of the respondents spent respectively three up to seven
days per week on chewing khat. These results indicate that the participants of the survey spent a
lot of resources on khat especially time and money. Hence exposure to pesticides from this crop
is likely to be the case.
7.3.2 Occurrence of DDT and its metabolites in khat
The results of DDT and its metabolites detected in khat samples are given in table 7.1. Based on
the results, p,p’-DDE, p,p’-DDD, p,p’-DDT and o,p’-DDT were detected in 80%, 70%, 61.7%
and 58.3%, of the khat samples, respectively. Total DDT which is the sum of its metabolites was
detected in 80% of the khat samples. The primary metabolites p,p’-DDE and p,p’-DDD were
detected in a large number of khat samples.
Table7.1. Occurrence of DDT and its metabolites in the khat samples.
Pesticides No of sample % < LOD No of sample %
ND Detected Detected
p,p’-DDE 12 20.0 48 80.0
p,p’-DDD 18 30.0 42 70.0
o’p-DDT 25 41.7 35 58.3
p,p’-DDT 23 38.3 37 61.7
Total DDT 12 20.0 48 80.0
ND = None Detected, LOD = Limit of Detection
The concentration of the total DDT in an increasing order of khat samples are indicated in figure
7.3. As it is noticed from the figure, some of the samples contain total DDT residue above the
MRL set by Codex Alimentarius (FAO/WHO, 2013), while about more than three fourth of the
khat samples contained a total DDT above the European Commission (EC) MRL (EFSA,
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2012b). This indicates that there was illegal use of DDT in the study area and/or contamination
from use for indoor residual spraying (IRS) in the framework of malaria control. Jimma zone is
indeed known as one of the malaria endemic areas of Ethiopia (Karunamoorthi and Hailu, 2014).
Figure 7.3. Concentration of total DDT in an increasing order of the khat samples. Lines =
Maximum residue limit (MRL)
The mean concentration of total DDT and its metabolites for six different sample sources in
southwestern Ethiopia are presented in table 7.2. From the results, the mean total DDT residue
in khat was generally higher in samples from Sekoru (71.70 µg/kg), Dedo (149.00 µg/kg), Saka
(137.00 µg/kg), Kersa (103.00 µg/kg) and Mana (73.00 µg/kg) districts except in the samples
from Gomma district which was relatively low (41.20 µg/kg) compared to other five districts.
Table 7.2. Concentration of total DDT and its metabolites (µg/kg) in khat from six sources
Sample Number of Mean concentration of DDT and its metabolites
source samples p,p’-DDE p,p’-DDD o,p’-DDT p,p’-DDT Total DDT
Sekoru 6 61.65 7.53 7.18 13.96 71.67
Dedo 6 113.28 7.72 6.89 25.97 148.87
Seka 5 64.13 13.66 8.35 18.83 136.84
Kersa 30 68.94 21.155 7.595 16.71 102.89
Mana 5 60.12 12.07 8.49 10.31 72.98
Gomma 6 33.35 11.20 7.75 10.19 41.24
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The concentration of DDT metabolites significantly (p-value < 0.0001) different in khat samples
figure 7.4. The metabolite p,p’-DDE is distributed more in khat samples and is contributing
more in the sum of the total DDT. The presence of more metabolite than the parent compound is
due to the historical use of DDT in the study area.
Figure 7.4. The concentration distribution of DDT metabolites in khat samples
7.3.3 Exposure assessment
Probabilistic exposure analyses were worked out to evaluate whether the level of exposure of
khat consumers exceeded the provisional tolerable daily intake (PTDI) of DDT. A total DDT
intake above the PTDI is considered as a health risk related to DDT for khat consumers.
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7.3.4 Probabilistic exposure analysis
Table 7.3 presents the probabilistic estimates of total DDT intake from consumption of khat. The
mean khat consumption for the total population was 19.590 ± 4.560 g/kg bw/day, while the mean
DDT concentration detected in khat samples was 0.10 ± 0.080 mg/kg khat. The mean estimated
daily intake (EDI) of total DDT from consumption of khat is 0.002 ± 0.003 mg/kg bw/day, while
p97.5 is 0.006 mg/kg bw/day. Both values are below the PTDI and indicated no health risk.
Table 7.3. Results from probabilistic exposure analysis for khat consumer
Distributions Mean StDv p50 p75 p90 p95 p97.5
khat consumption
19.585 4.561 19.206 22.024 25.147 27.461 29.846
(g/kg bw/day)
DDT concentration
0.100 0.080 0.085 0.142 0.208 0.255 0.301
(mg/kg)
EDI
0.002 0.002 0.002 0.003 0.004 0.005 0.006
(mg/kg bw/day)
EDI = estimated daily intake
7.4 Discussion
From the results, 72% of khat chewers spent more than fifty Ethiopian birr per day to buy khat
and 50.7% of respondents spent more than three days per week on chewing khat. This may be
due to negligence of the participants, awareness of problems related to khat chewing and
influence of peers who motivate them to chew khat. Additionally, the use of khat is resource
intensive in terms of time, money and human power. Creating awareness about the benefits and
risks of khat is important to protect consumers. From the Mann Whitney U Test, the distribution
of average khat consumption (g/kg bw/day) is the same in male and female chewers (p-value >
0.05). This indicates that there may be similar exposure for both female and male khat chewers
to DDT.
In the present monitoring study, results showed that a high incidence of DDT in/on the chewable
parts of khat coming from all investigated areas. As reported in figure 7.3, more than three
fourth of the khat samples contain total DDT concentrations above the EC MRL which is set for
every agricultural crop worldwide by the European Food Safety Authority (EFSA, 2012b) and
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also other organochlorine pesticides in a variety of vegetables and fruits are set by default at 0.05
µg/kg (Zhang et al., 2011). The presence of DDT in khat above the MRL indicates that there was
illegal use of DDT in the study areas and/or contamination of the khat samples from the use of
DDT for indoor residual spraying (IRS) in the framework of malaria control. Jimma zone is
indeed known as one of the malaria endemic areas from Ethiopia where DDT is applied to
control the vector (Karunamoorthi and Hailu, 2014). The occurrence of DDT in khat can also be
explained by its persistent nature as a pollutant in the environment as has been discussed in
chapter three and four.
From Kruskal-Wallis test the difference in the concentration of total DDT between the khat
sample sources (six districts of Jimma zone) was not significant (p-value = 0.423). Compared
with khat samples collected from different parts of Ethiopia such as Hararge and Butajira (Daba
et al., 2011), the mean residue of total DDT detected in the present study was lower. When
compared with the study done in Sidama zone (Ligani and Hussen, 2014), higher total DDT
residues were detected in the present study for all khat samples. This revealed that there was
intensive application of DDT in our study area compared to Sidama zone. After application of
DDT, it stays long in the environment with a half-life of 2 to 15 years due to its persistent nature
(Mahdavian and Somashekar, 2013). From the post-hoc analysis, the concentration of the
metabolite p,p’- DDE is highly distributed in the khat samples. Several literatures (EFSA, 2006;
Kalantzi et al., 2001; Sifuentes Dos Santos et al., 2015) report that a high accumulation of p,p’-
DDE and p,p’-DDD in environmental samples indicate that there was historical use of DDT. The
parent DDT gets metabolized over time to p,p’-DDE and p,p’-DDD. From this it can be
concluded that the contamination of khat in Jimma zone may be due to historical use of DDT. It
may also originate from the dumped obsolete pesticides mostly containing organochlorine
pesticides such as DDT in large amounts as reported by Daba et al. (2011). Due to this dumping,
pesticides may leak from the storage areas to the different environmental compartments and
contaminate agricultural products such as khat.
The probabilistic exposure analysis showed that the mean concentration of total DDT exceeded
the maximum residue limit (MRL) set by the Codex Alimentarius for different agricultural food
items which is 0.1 mg/kg (FAO/WHO, 2013). This indicates that there is a high contamination of
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the khat leaves by total DDT. From the results of the exposure analysis, the mean and p97.5 EDI
was below the PTDI set by Codex Alimentarius. But this does not guarantee the safety of khat
chewers. As stated in the socio-demographic results of the present study, around 25% of the
participants chew khat seven days per week. A continuous exposure to DDT results in
bioaccumulation of this pesticide in the body, which may result in chronic health problems.
Additionally, the khat consumers may have cumulative exposure to DDT through consumption
of other commonly consumed agricultural crops in Ethiopia like teff, maize, red pepper which
also contain DDT and its metabolites as reported by previous study (Mekonen et al., 2014).
7.5 Conclusions
The present study investigated consumer exposure to DDT from khat consumption in
southwestern Ethiopia. From the results of the study, most of the khat consumers were male.
However, there is no significant difference in the pattern of the amount of khat consumed. This
indicates that there is similar exposure to DDT between the male and female population. From
the analysed khat samples, 80 percent contained DDT residues and some of the residues were
above the MRL set by the Codex Alimentarius and most of the residues were above the EC MRL
for DDT, indicating illegal use of DDT. Among the metabolites, p,p’-DDE was detected in large
numbers of the khat samples compared to the parent compound p,p’-DDT. This indicates that the
contamination of the khat samples comes from historical use of this pesticide. The presence of
DDT and its metabolites in khat is posing an additional health risk to regular khat chewers. From
the probabilistic exposure assessment, the mean and p97.5 EDI of DDT was below the health
based reference value (PTDI) and indicated no risk for the khat consumers. However, this does
not guarantee the safety of the khat consumers, because they may be exposed to DDT from other
food sources. Above that the khat plant is consumed raw without any preceding processing.
Therefore, continuous monitoring of pesticides including DDT in khat and other commonly used
agricultural crops is recommended to sustain the safety of consumers.
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Chapter Eight: General discussion, conclusions and future research prospects
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8.1. General discussion
Despite the popularity of pesticides and their extensive use, serious concerns come up about
health risks arising from the exposure of farmers, when mixing and applying pesticides, or
working in treated fields and from exposure to residues in food and in drinking water ( Maroni et
al., 2006; Lopes Soares and Firpo de Souza Porto, 2009). This exposure of consumers may result
in either long or short term health risks.
Environmental contamination of farmland and water in Ethiopia is mainly due to the use of
pesticides in public health to control vector borne diseases and in agricultural sector to reduce
pest problem and/or eliminate crop yield losses. For both agricultural and public health uses of
pesticides, the targets are the pests and the diseases. However, human beings can be exposed to
these chemical pesticides due to consumption of treated fruits, vegetables and cereals or due to
pesticide contaminated drinking water. Problems related to pesticide residues are severe in
developing countries, where there are no sufficient skills, no awareness, and very poor law
enforcements (Yáñ ez et al., 2002).
8.1.1. Pesticide residues problems in foods
What is the problem related to pesticide use in developing countries, particularly in Ethiopia
(RQ1)? Does the commonly consumed food commodities produced in southwestern Ethiopia
contain pesticide residues (RQ2)?
The use of pesticides in Ethiopia is a common practice as explained in chapter two; however,
this is at the expense of the environment and human health. Most of the farmers are illiterate. As
a result they apply pesticides in violation with the recommendations because they do not
understand the labels on pesticide containers, they use unsafe storage practice, they do not know
the proper dose or concentration to apply, ignore risks and safety instructions, do not use
protective devices during application of pesticides and dispose pesticide containers unsafely or
they may use empty containers for drinking water or to store food in it (Mengistie et al., 2015).
Such problems aggravate environmental contamination resulting in human exposure. To
understand the risk of human exposure to pesticides, the residue at different sources, such as
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food, water and other consumer products should be known. Although there are a limited number
of studies available dealing with pesticide residues in Ethiopia ( Geletu et al., 2009; Daba et al.,
2011; Gebremichael et al., 2013), no study evaluated the level of human exposure to pesticides
from consumption of agricultural produce and drinking water. Additionally, in Ethiopia,
pesticide monitoring programs for import products, about safe handling and human exposure to
pesticides, in particular from food consumption, are not well coordinated by the government. The
main focus of the government and people in Ethiopia is to secure food and is not dealing with
risk/benefit analyses for pesticide use. Moreover, there was no study undertaken on market
samples of agricultural produce which represent the majority of Ethiopian consumers.
Taking these gaps into account, this PhD study is the first in Ethiopia which addresses pesticide
residues in different food items, khat, drinking water sources and human exposure based on the
amount consumed. This study concentrated on evaluation of pesticide residues in major staple
food items (commonly consumed foods) such as teff (Eragrostis tef), maize (Zea mays), red
pepper (Capsicum annuum), coffee (Coffea arabica), lentil (Lens culinaris) and wheat (Triticum
ssp.) (chapter three). The results of this study indicate that the commonly consumed food items
were contaminated with different pesticides. This contamination may come from the use of
pesticides in agriculture or public health and may be from the presence of dumped obsolete
pesticides in different areas in Ethiopia. Pesticides which are no more in use for agriculture such
as DDT and endosulfan were also detected in these food samples. Although, the use of DDT as
an insecticide for agricultural use has been restricted since 1981, and was banned from the
market since 1986 in Europe, DDT is still used in some developing countries until recent years.
The results of our study confirmed that there was recent use of DDT, as the parent molecule p,p’-
DDT was detected more than its metabolite p,p’-DDE in the staple foods.
The extended use of DDT as vector control for malaria is still recommended by WHO (EFSA,
2006), however this may also cause cross contamination to food, drinking water or other
consumer products. The detection of DDT in the environment is a serious problem due to its
persistent nature, the abilities to biomagnify in the food chain and bioaccumulate in the human
body (Tomza-Marciniak and Witczak, 2009).
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The concentrations of a couple of pesticides in the staple food items were above the MRL set by
Codex Alimentarius (FAO/WHO, 2013). This may be due to the fact that most of the farmers in
Ethiopia are not aware of and do not respect the legal requirements for the use of pesticides
either for agricultural application or for other purposes. Moreover, the cause of higher pesticide
residues detected in commonly consumed food items may be due the use of poor application
techniques and equipment (Mekonnen and Agonafir, 2002).
8.1.2. Human exposure and associated risk to consumers
Are people exposed to different types of pesticides detected in the agricultural crops (RQ3)?
How safe is the level of human exposure to pesticides from food, water and khat consumption
(RQ4)? Do pesticides cause a health risk for short and long term exposure when residues are
compared to health based reference values (RQ5)?
As has been explained in the introduction chapter human exposure to pesticides happens due to
their occupation or from consumption of pesticide contamineted food. Infants are specific groups
of the population who may suffer more from the risk of pesticide exposure. As an example, our
study (chapter four), investigated infants exposure and health risks to DDT residue in maize
used as major component of their food. From the risk estimation results, the mean and the 97.5
percentile estimated daily intake (EDI), were above the provisional tolerable daily intake (PTDI)
of DDT set by Codex Alimentarius (FAO/WHO, 2013). This indicates a chronic health problem
for the infants taking maize as a complementary diet ((EFSA, 2006). This looks problematic for
most African countries, as they have a higher uptake of DDT compared to Europe and America
where no exposure to these kind of compounds takes place (Jaga and Dharmani, 2003).
Does contamination of drinking water with pesticides cause a health problem to consumers
(RQ6)?
Human beings are not only exposed to pesticides in their food but also from the water they drink.
By the fact that most of the water bodies are at risk of pesticide contamination as they are mostly
located near to the agricultural areas. Water contamination results from the release of pesticides
from field run off or effluent discharges and water sources such as rivers, dams, ponds, springs,
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wells etc. are the immediate pesticide receivers (Chee et al., 1996). From our study (chapter
six), water sources in Jimma zone and Addis Ababa were found to be contaminated by
organophosphate pesticides (malathion, diazinon and pirimiphos methyl), a herbicide (2,4-D)
and a fungicide (fenpropimorph). These pesticides were also detected in tap water and in
distribution reservoirs. This indicates that the pesticides pass the conventional water treatment
plant and appear until the water is ready for consumption.
Most water treatment facilities in the country are mainly focused on the removal of
microorganisms, turbidity, and conductivity but do not consider chemical hazards such as
pesticides. Conventional water treatment methods such as particle coagulation-flocculation,
sedimentation and filtration are not 100% effective for the removal of pesticides unless there is a
possibility to apply other advanced water treatment methods such as nano-filtration, reverse-
osmosis (Karabelas and Plakas, 2011). As a result, urban communities are exposed to pesticides
in drinking water. Moreover, the majority of the rural communities of Ethiopia who have no
access to conventionally treated water sources are even exposed to higher concentrations of
pesticides as they take water directly from the surface or ground water sources.
The frequent detection of pesticide residues in water indicate potential adverse effects to
consumers even at very low concentrations (McKinlay et al., 2008; Sanborn et al., 2004).
Pesticide exposure assessment in our study (chapter six) indicates that the estimated daily
intake (EDI) from drinking water of some pesticides was above the health norm, particularly the
chronic acceptable daily intake toxic parameter (EDI > ADI). This indicates that urban and rural
communities of Ethiopia may face chronic health problems from pesticides in drinking water.
While, all pesticides detected in drinking water sources were below the acute reference dose
(EDI < ARfD). This may be due to the low concentration in the water which may not cause acute
risk as it depends on the dose or concentration of the chemical. Pesticides found in water
supplies were not usually present in high concentrations causing acute toxic effects (Trautmann
et al., 2012).
The build up of buffer zones around the water sources and also stewardship and frequent
monitoring of the pesticide application strategies, selecting the type of pesticides, and also
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advancing the water treatment technology in Ethiopia can help to minimize problems related to
pesticide exposure from drinking water.
Is a human exposure to pesticides happen due to consumption of conventionally non-edible plant

such as khat (RQ7?
The route of human exposure to pesticide is not only drinking water or conventionally edible
foods, but also other consumer products such as khat (Catha edulis). Khat is a raw plant material
consumed in many parts of East Africa. Even though, the use of khat is restricted in most
European countries, it’s common in countries located in the horn of Africa, such as Ethiopia. In
Ethiopia, this plant is widely consumed regardless of man or women. Chewing khat increases the
release of active constituents cathine and cathinone which causes loss of appetite (Murray et al.,
2008). In some regions of Ethiopia where there is shortage of other crops, people believe that
consuming khat is an important means to alleviate hunger and they consider it as food (personal
observation). However, this plant may also be contaminated by pesticides which may cause a
health risk to human. Our study (chapter seven) indicates that DDT and its metabolites were
present in khat plant. The metabolite p,p’-DDE was detected at a high concentration and in a
large number of samples, which indicates historical use of DDT. From the result of human
exposure to DDT in khat, the EDI was less than the health based reference value (EDI < ADI).
However, this does not guarantee 100% safety as the plant is consumed fresh without any
preceding household processing steps. The presence of pesticides in khat should receive more
attention from the government, community and also academia in order to be able to minimize the
risk.
8.1.3. Cumulative risk assessment for multiple pesticides detected in commonly

consumed foods
Is there a cumulative risk for consumers from mutual consumption of different food items
(maize, teff, red pepper, coffee and khat) and drinking water in Ethiopia (RQ9)?
Ethiopian population may be exposed to multiple pesticides due to mutual consumption of food,
drinking water and khat. Therefore, addressing the cumulative risk is interesting in this area.
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Dietary risk assessment of the human exposure is traditionally performed for a single pesticide
and a single crop. However, a food consumed daily may contain more than one pesticide or
people may consume a combination of food items containing different pesticides (Quijano et al.,
2016). If the detected pesticides have the same mechanism of action or the same toxicological
endpoints, assessing the dietary risk for individual pesticides may underestimate the health risk
(Boon et al., 2008). To address the combined effect of the compounds, undertaking cumulative
risk assessment (CRA) is important. Cumulative risk assessment is defined as, the simultaneous
exposures to various chemicals which contribute to a cumulative effect in the human body. The
combined effects to humans can have additive, response addition or interaction either synergistic
or antagonistic effects (European Food Safety Authority (EFSA), 2008). The combined hazard of
the individual compounds in cumulative assessment groups (CAGs) can be determined using
either the reference point (RPs also known as ‘point of departure’ (POD), or the ‘reference
values’ (RVs) such us health based guidance values for example ‘acceptable daily intake’ (ADI)
and the ‘acute reference dose’ (ARfD). Currently, there is no internationally agreed methodology
to assess risks from multiple chemicals.
However, to assess the exposure to pesticides as a mixture, the first step is to identify the
substance with a common mechanism of toxic action (CMGs) (Gallagher et al., 2015). For the
assessment of the cumulative risk from exposure to CMGs, the ‘relative potency factor’ approach
(RPF), based on dose addition is the appropriate method (Jensen et al., 2013; Kennedy et al.,
2015).This approach assumes that doses of component chemicals, which act in a toxicologically
similar way, can be added after scaling the doses by their potencies relative to an index chemical.
The RPF approach is transparent and relatively easy to understand because potency correction is
separated from consideration of exposure. As a consequence, it provides an effective means for
standardizing the dose metrics for the toxicity of the different compounds of the cumulative
assessment group (CAG) (Boobis et al., 2008). The risk is assessed after comparing the
cumulative dietary exposure with a toxicological reference dose such as the ADI of the index
compound in case of chronic exposure and the ARfD of the index compound for acute exposure
(Quijano et al., 2016).
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In our study (chapter three), pyrethroids such as deltamethrin, cypermethrin , permethrin and
the organochlorine DDT were detected in commonly consumed food commodities such as maize
(Zea mays), teff (Eragrostis tef), red pepper (Capsicum annuum) and coffee (Coffea arabica).
DDT was also detected in khat (Catha edulis) (chapter seven). According to US environmental
protection agency, pyrethroid and organochlorine pesticides have common mechanism of action
(CMGs). They are neurotoxic chemicals acting on the axonic terminal of the central nervous
system and causing behavioural changes for the exposed people (USEPA, 2011). Ethiopian
people are exposed to these CMGs, through mutual consumption of the above food commodities,
drinking water and khat containing multiple pesticides. Therefore, assessing the cumulative risk
to these multiple pesticides is important to assure consumer safety. In dietary risk assessment,
consumption and pesticide residue data’s are always needed. For this study, only consumption
data for maize (Mekonen et al., 2015) and khat (chapter seven) are available, while for other
food commodities; the mean national annual per capita consumption is considered i.e., teff (25.9
kg) (Berhane et al., 2011), coffee (2.4 kg) (Zeru, 2006), and red pepper (0.62 kg) (CSA, 2001).
From annual per capita consumption, the daily consumption of each food items was calculated
(annual per capita consumption/365 days) and expressed in g/kg bw/day.
Food processing such as roasting, backing, peeling, cooking and others may have an effect on the
concentration of pesticides and on level of human exposure as well. Therfore, for our cumulative
dietary risk assessment, the processing factor (PF) was taken into account for teff and coffee. For
red pepper and maize no data on the processing factor were available. The risk was assessed by
assuming that processing had no effect on the residue of the pesticides under study (worst case
scenario). Khat is consumed raw without any preceding processing and it is considered as it is.
After assessing the cumulative exposure, the results were compared with the health based
reference values such as the ADI and ARfD to assure food safety. For our study, the relative
potency factor is determined based on the ADI of the three pyrethroids (cypermethrin,
permethrin and deltamethrin) 0.02, 0.05 and 0.01 mg/kg bw/day, respectively and one
organochlorine (DDT) with an ADI/PTDI of 0.01 mg/kg bw/day (FAO/WHO, 2013). From these
pesticides, deltamethrin was chosen as an index compound due to the fact that its ADI value is
five times lower compared to permethrin and two times lower compared to cypermethrin.
Deltamethrin is also used as an index compound in another study (Quijano et al., 2016).
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The deterministic calculation (low tier approach) was undertaken to assess the cumulative
exposure from this multiple chemicals. The individual estimated daily intake (iEDI) for a
pesticide in a food commodity was calculated as follows:
iEDI = Ci*Fi
Where:
Ci: residue level in food commodity
Fi: food consumption of food commodity
The cumulative exposure was expressed as deltamethrin equivalents by multiplying the iEDI of
each pair pesticide-commodity by the relative potency factor (RPF) of deltamethrin and adding
up to one cumulative intake i.e., ⅀iEDI*RPF.
The results of the cumulative exposure are indicated in table 8.1. The result of this assessment
indicates that the cumulative exposure to pyrethroid and organochlorine pesticides for each crop
were above the ADI set by JMPR of FAO/WHO (FAO/WHO, 2013) except for those detected in
coffee beans and khat. This reveals that the population in southwest Ethiopia will have chronic
health problems from multiple pesticides due to mutual consumption of teff, red pepper and
maize. The cumulative exposure from coffee is less than ADI, this may be due to the application
of the processing factor as most of the pesticides disappear due to household processing
(Mekonen et al., 2015). This result is consistent with the results obtained in China, on the long
term cumulative risk for organochlorine and pyretroids from consumption of three types of nuts
for which the results were above ADI (Liu et al., 2016). On the contrary, the study done in Spain
indicates that the chronic cumulative intake of a pesticides through fruits and vegetables were
below ADI and no cumulative chronic consumer risk is expected (Quijano et al., 2016).
However, the pyrethroids and the organochlorine (DDT) were not detected in drinking water
sources from both sampling points (Jimma and Addis Ababa). This may be due to the fact that
these pesticides are not water soluble, while highly lipid soluble (Davies et al., 2007). For this
assessment drinking water have no effect. Total cumulative exposure is 0.526 mg/kg bw/day,
which is by far greater than the ADI and there will be a chronic health problem for consumers.
140
Table 8.1.Cumulative exposure to pyrethroid and organochlorine pesticides through consumption

of staple food items, khat and drinking water
Cumulative
Mean risk
Upper bound consumption iEDI (iEDI*RPF) ADI
Food concentration (g/kg (mg/kg (mg/kg (mg/kg
commodity Pesticides (mg/kg) bw/day) bw/day) bw/day) bw/day)
Teff cypermethrin 0.700 70.960 0.050 0.100
permethrin 0.481 0.034 0.171
DDT 0.444 0.031 0.031
⅀iEDI*RPF 0.302 0.01

Red
pepper cypermethrin 0.618 0.001 0.006
permethrin 2.154 0.004 0.022
DDT 0.868 2.000 0.002 0.002
⅀iEDI*RPF 0.031 0.01

Coffee deltamethrin 0.348 0.002 0.002
6.580
DDT 0.247 0.007 0.007
⅀iEDI*RPF) 0.009 0.01

Maize cypermethrin 0.428 0.017 0.087
permethrin 0.373 0.015 0.076
40.540
DDT 0.416 0.017 0.017
⅀iEDI*RPF 0.182 0.01

Khat DDT 0.102 19.580 0.002 0.002
⅀iEDI*RPF 0.002 0.01

Drinking water - - - -
Total ⅀iEDI*RPF 0.526 0.01
RPF = Relative Potency Factor, iEDI = individual Estimated Daily Intake, ADI = Acceptable
Daily Intake, values above ADI indicated in bold
From the food commodities, teff contributed more to the cumulative exposure followed by maize
(figure 8.1). This may be due to the high consumption level of these two cereals compared to
others. In Ethiopia most of these food commodities are consumed together in one day. For
example, one may eat injera (traditional flat bread) made from teff or maize with red pepper
spiced sauce. Additionally, every day coffee is drunk and khat consumption is also common
141
(personal observation). Therefore, people are highly exposed to multiple pesticides from mutual
consumption of these food commodities.
0.009
0.002
0.182
Teff
Red pepper
Maize
0.302 Coffee
Khat
0.03
Figure 8.1. Cummulative risk (mg/kg bw/day) of multiple pesticides from mutual consumption
of the above consumer products (mg/kg bw/day)
Therefore, creating awareness to pesticide users and application of integrated pest management
(IPM) to minimize the use of pesticides should receive more attention in Ethiopia to curtail the
cumulative risk.
Our findings in the cumulative risk assessment may have some limitations. The consumption
data we used for the calculation of the cumulative exposure were from a single annual per capita
consumption, due to the absence of enough consumption and pesticide residues monitoring data.
This has hindered to work out a probabilistic cumulative risk assessment. Hence, deterministic
(point estimate) evaluation was applied considering the mean of the estimated daily intake in the
calculation. These may result in under or overestimation of the results as reported by different
researchers (EFSA, 2012a; Nougadère et al., 2012; Quijano et al., 2016). In this regard, more
effort has to be done for the collection of national consumption data and residue monitoring not
142
only for these pesticides but also for other commonly used pesticides in agriculture is
recommended to undertake a more comprehensive risk assessment.
8.1.4. Importance of food processing for the reduction of pesticide residues in food
Does household processing such as washing, roasting, brewing, and doughing and baking have
an effect on residues of pesticides in raw food items (RQ7)?
To avoid food safety issues related to pesticides, organic farming is being propagated in the
world. However, organic farming is not well practiced in most developing countries for several
reasons, such as the increasing population size, lower yields from agricultural production due to
the effect of pests, low awareness of the pesticide users on organic practices and wrong
perception of the farmers about pesticides. Most farmers in Ethiopia considering pesticides are
the best option to increase agricultural production. In addition to this, there are critics that
organic farming use more land compared to conventional agriculture (Seufert et al., 2012).
Therefore, it is still important to look for other solutions to tackle food safety problems related to
pesticide residues in agricultural crops and drinking water.
One advantage for consumers is that food processing minimizes pesticide residues in food which
had been shown in this work. The food processing operations such as washing, peeling, cooking,
blanching and baking play a great role in the pesticide residue reduction in food (Elkins, 1989).
As a result, food processing at household and industrial level improve the current scenario of
unsafe food (Kaushik et al., 2009).
Our study in chapter five indicates that household processing of coffee and teff reduces
pesticides residues from the raw commodity. Even the high thermal resistant pesticide, DDT and
its metabolites showed a significant decrement particularly during the processing of coffee. This
was mainly due to the frequent exposure of the coffee to heat during the roasting and brewing
processes. Baking is also the most effective household processing for the reduction of pesticides
in teff. The PFs for both crops were found to be less than one, which is an indication of a
reduction of pesticide residues compared to the raw commodity. The percent reduction varies
from processing to processing. The process with the application of heat such as roasting, brewing
143
and baking has the greatest impact on the pesticide residue reduction. The effects of food
processing is influenced by the physicochemical properties of pesticides such as the solubility,
thermal degradation, octanol-water partition coefficient and volatility (Holland et al., 1994;
Kaushik et al., 2009).
In conclusion, stewardship to enforce the law and create awareness by the government towards
the pesticide users, strict monitoring, processing of food before consumption and risk evaluation
are important tools to sustain consumer safety of the Ethiopian population.
8.2. Conclusions
While pesticides are important for the control of pests, they may cause side effects on human
health and the environment. Developing countries, in particular Ethiopia suffers from problems
related to pesticides use. Despite this fact there are little attention on potential research areas,
lack of law enforcement or poor policy directions and relevant actions to at least minimize the
risks related to pesticides. The goal of this dissertation was to identify problems related to
pesticide contamination in agricultural produce, water, khat, human exposure due to
consumption and to assess the effect of food processing on level of pesticides in food. These
makes a novel contribution to the literature by demonstrating that human exposure can be
associated with the consumption of pesticide contaminated food, drinking water and other
agricultural crops which pose a threat to health.
To sumerize the key issues, the following conclusions are drawn based on the findings of the
PhD study:
 It can be concluded that food items, water and khat are often contaminated with
pesticides that may arise from agricultural and public health use of pesticides and the
presence of obsolete stocks in Ethiopia (answered RQ2, RQ6 and RQ7).
 Some pesticide residues in the foods were above the legal reference doses which reflect
the maximum allowable concentration in food. This indicates that good agricultural
practice (GAP) is perhaps not be well practiced in Ethiopia (answered RQ1 and RQ2).
144
 From the results of human exposure and consumer risk assessment, it is concluded that
human beings, in particular infants are exposed to non-authorized pesticides such as DDT
from consumption of their complementary diet (answered RQ3, RQ4 and RQ5).
 Consumers are daily exposed to pesticides from drinking water, in which the
contamination may come from the surrounding environment (answered RQ4, RQ5 and
RQ6).
 Khat, being a stimulant plant which has naturally an impact on consumer’s health, is also
contaminated with pesticides such as DDT. This could pose a double burden on human
health (answered RQ4, RQ5 and RQ7).
 Mutual consumption of food items, which are contaminated with multiple pesticides
belonging to a common mechanism of action group (CMGs), shows a cumulative health
effect on the Ethiopian population (answered RQ 9 and RQ10).
 In general, developing a continuous pesticide monitoring program, law enforcement such
as on time implementations of policies and regulations, networking within different
stakeholders, stewardship and creating awareness to pesticide users, promoting the use of
integrated pest management (IPM) in Ethiopia are important to minimize problems
related to human exposure to pesticides and associated health risks.
 Strong regulation regarding pesticides import, use, storage and continued pesticide
monitoring in food and other environmental samples seems vital to assure the safety of
consumers in Ethiopia.
The following roadmap (figure 8.2) indicates proposed actions at different levels of the chain
to minimize the problem related to human exposure to pesticides in Ethiopia.
Micro-level: at this level farmers and communities who are working to secure food should
act. These group of the production chain (chain function stage) need support to improve their
production systems and update their knowledge on pesticide handling to minimize pesticide
related risk.
Meso-level: scientific people, NGO’s and different associations who support both micro and
macro levels (chain supporters). These groups can support financially or giving scientific and
policy advice.
145
Macro-level: decision makers or enablers/regulation of the processes in the chain, such as

policy makers, implementers of the regulations, giving certifications and others.
If the use of pesticides causes human exposure and health risks, what will be the next step
(RQ10)?
146
Important actions for improvement of problems related to pesticide contamination in crops, water and human
exposure in Ethiopia
MICRO LEVEL
(Farmers and Communities)
Support
 Create awareness on benefits and risks of pesticides
 Adopt safer ways of handling, storing and disposing of pesticides
 Use of personal protective equipment
 Change perception regarding pesticides
 Inspection of pesticide application procedures
 Follow up of the pesticide suppliers to farmers
 Distribute list of authorized pesticides for each crop
 Establish documentation system on pesticide usage

MESO LEVEL
(Research Institutions, Academians, NGOs, Farmers Associations and Civil Society)
Research Support
 Provide scientific advice for risk managers  Lobby with the government to enforce the regulation and food safety
 Undertake detailed research in every aspect related to pesticide management systems
 Propose other opportunities to minimize pesticide use  Propose means on local management of obsolete pesticides
 Provide workshops and trainings regarding the management of safety risks  Assist farmers and food exporters in laboratory tests for agricultural
 Give especial attention to exposure of vulnerable groups or YOPI’s (Young, produce
Old, Pregnant women and Immune compromised)  Create awareness about selection of play fields for children
 Communicate their research findings
MACRO LEVEL
(Government (FDRE), Regional Governments (Nine regional governments) and Regulatory Institutions (Eg. MoA, FMHACA)
Legal Framework Networking

 Enforce the regulations regarding pesticide registration and control  Establish strong networking with all stakeholders
 Develop legal requirements for import and local formulation of pesticides  Create risk awareness to general public
 Establish national maximum residue limits for agricultural produce  Lobby donor countries and local NGO’s to establish a program for disposal
 Establish proper pesticide monitoring programs of obsolete pesticides
 Broaden up analytical capacity for every agricultural crop not only coffee
 Develop national guidelines and standards regarding pesticides in different Research
environmental matrices
 Establish research centers and develop institutional set up
 Strict control of illegal retailers of pesticides and their sources
 Encourage researchers to report their findings in different medias
 Provide appropriate funding for research and capacity building
 Establish a system for the accessibility of data’s regarding pesticides in
Ethiopia
*FDRE = Federal Democratic Republic of Ethiopia, MoA = Ministry of Agriculture, FMHACA = Food, Medicine
and Health Care Administration, and Control Authority, NGOs = Non-governmental Organizations
Figure 8.2. Specific recommendation for intervention at three levels of chain to minimize
pesticide related problems in Ethiopia
147
8.2. Future research prospects
The study deals about human exposure and risks to pesticides from consumption of food,
drinking water and other consumer products. The contamination of the pesticides may not be
limited to food and drinking water. Further research in this area should focus on the other
environmental compartments such as soil, air and biota living in the aquatic and soil
environment. Detailed analyses of pesticide residues in these environmental compartments
followed by environmental risk assessment (ERA) is recommended, particularly for those
pesticides which are able to persist in the environment and have the ability to biomagnify
through the food chain.
This study focuses on the estimation of human exposure based on residues and consumption
data; this does not indicate the real amount of uptake of these chemical pesticides in the general
blood circulation and/or target organs or tissues. Therefore, further study is needed using
biomarkers to identify the biological and toxicological available fractions of pesticides in the
human body.
In Ethiopia, there is no national consumption data base, for this study we used a consumption
survey on the 24-hr recall method for the assessment of the daily consumption by infants and
khat consumers. However, this might not give all the details of the usual dietary intake, so
further research and new food surveys about the Ethiopian diet over more days of consumption
representing the general population is of interest.
This study considers that the feeding status of most Ethiopian people is quite similar. However,
there are some cultural differences, so undertaking a national consumption survey based on the
different seasons and assessing human exposure to pesticides in all regions of the country is
needed to evaluate the pesticide exposure of the general population in the country.
Food processing has an effect on the residues of pesticides detected in the study for two
commonly consumed food items (teff and coffee). Further study is required to include also the
effect of other household and industrial food processing steps among different food items and for
other pesticides present.
148
Pesticide residue analyses is important to crop protection, research, environmental monitoring,

public health protection and legislative enforcement, to be conversant with the legislations of
importing countries, to be a competitive exporter of agricultural products, and to participate
effectively in international and regional trade. Therefore, future work should also be focused on
the establishment of a certified analytical laboratory for pesticide residue analysis in food and
other environmental matrices in the region according to internationally accepted quality
assurance (QA) and quality control (QC) procedures.
This study identifies the commonly applied pesticides regardless of how they are applied for
different purposes; further research is needed to monitor the spray habits of Ethiopian farmers
and pesticide applicators in general. This has to create of awareness in the community by giving
information on the good agricultural practices and the application of the right dose and right
pesticide for a specific crop pest of disease.
Finally future work has to be done, to create awareness to the regulatory bodies and decision
makers, about the problems related to pesticide residues in food and other environmental
samples, means of transport of these pesticides in the environment (run-off, drift and
volatilisation), problems related to the accumulation of obsolete stocks (control of the local
formulation and importation) and the importance of continuous pesticide monitoring to enforce
the application of integrated pest management (IPM) and pesticide regulations in Ethiopia.
149
150
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Zeru, A., 2006. Diversity of arabica coffee populations in afromontane rainforests of Ethiopia in
relation to collectotrichum kahawae and giberella xylarioidses.
Zhang, W., Jiang, F., Ou, J., 2011. Global pesticide consumption and pollution: with China as a
focus. Proc. Int. Acad. Ecol. Environ. Sci. 1, 125–144.
174
Annexes
Annexes
175
Annexes
176
Annexes
Annex 1: Model out puts from probabilistic exposure analysis of DDT in khat
Fit Comparison for Khat consumption (g/kg bw/day ...

Riskloglogi.stic(-6,8555;26,062; 10,7000
12,50 28,45
90,000
92, ...
0,12
0,10
• Input
Minimum 8,190
0,08 Maximum 33,000
Mean 19,580
Std Dev '1,'152
Values 423
0,06
- Loglogislic
0,04 Minimum -6,856

Maximum +co
Mean 19,585
Std Dev '1,561
0,02
0,00
....
0
I
~ 0 Ln
Probability-Probability Plot of Khat consumption (g/kg bw/day...

Riskl..oglogistic(-6,8555; 26,062; 10,7 000
1,0
0,9
0,8
0,7
_j 0,6
~c.. 0,5 - LogLogistic
1:J
IV
;:! 0,4
lL
0,3
0,2
0,1
0,0
0 .... N 1\"1
è o' o' o'
Input p-Va ...
177
Annexes
Fit Comparison for DDT concentratien mg/kg k...

RiskExtvalue(0,064046;0,0618...
0,0100 0,2432
10
9
8 • Input
Minimum 0,01000
7 Maximum 0,37201
Mean 0,10204
6 Stel Dev 0,08681
Values 60
5
4 - ExtValue
Minimum -oo
3 Maximum +oo
Mean 0,09976
2 Stel Dev 0,07936
0
1/) 1/) 1/)
0
0
0 0 .....
0
.....
9 è è è è
Probability-Probability Plot of DDT concentratien mg/kg k...

RiskExtvalue(0,064<l46;0,0618 ...
1,0
0,9
0,8
0,7
j 0,6
~
0.. 0,5 - ExtValue
"~ 0,4
LL
0,3
0,2
0,1
0,0
.....
0
è o·
N
è
'lf".
o
Ltt
o
\0
o·
0
.....·
Input p-Va ...
178
Annexes
Annex 2: @risk detailed statistics for the intake of DDT due to consumption of khat from Monte-Carlo simulation
model
Intake of DDT in
Intake of DDT in khat Intake of DDT in khat Intake of DDT in khat Intake of DDT in khat
khat (mg/kg bw/day)
Summary statistics (mg/kg bw/day) Output (mg/kg bw/day) Output (mg/kg bw/day) Output (mg/kg bw/day) Output
Output (Sim#1)
(Sim#2) Sheet1!G8 (Sim#3) Sheet1!G8 (Sim#4) Sheet1!G8 (Sim#5) Sheet1!G8
Sheet1!G8
Minimum -0.0023 -0.0023 -0.0023 -0.0023 -0.0023
Maximum 0.0211 0.0211 0.0211 0.0211 0.0211
Mean 0.0019 0.0019 0.0019 0.0019 0.0019
Std Deviation 0.0017 0.0017 0.0017 0.0017 0.0017
Variance 0. 0001 0. 0001 0. 0001 0. 0001 0. 0001
Skewness 1.4735 1.4735 1.4735 1.4735 1.4735
Kurtosis 7.1615 7.1615 7.1615 7.1615 7.1615
Errors 0 0 0 0 0
Mode 0.0010 0.0010 0.0010 0.0010 0.0010
5% Perc -0.0007 -0.0007 -0.0007 -0.0007 -0.0007
10% Perc 0.0002 0.0002 0.0002 0.0002 0.0002
15% Perc 0.0004 0.0004 0.0004 0.0004 0.0004
20% Perc 0.0006 0.0006 0.0006 0.0006 0.0006
25% Perc 0.0008 0.0008 0.0008 0.0008 0.0008
30% Perc 0.0009 0.0009 0.0009 0.0009 0.0009
35% Perc 0.0011 0.0011 0.0011 0.0011 0.0011
40% Perc 0.0013 0.0013 0.0013 0.0013 0.0013
45% Perc 0.0015 0.0015 0.0015 0.0015 0.0015
50% Perc 0.0016 0.0016 0.0016 0.0016 0.0016
55% Perc 0.0018 0.0018 0.0018 0.0018 0.0018
60% Perc 0.0020 0.0020 0.0020 0.0020 0.0020
65% Perc 0.0022 0.0022 0.0022 0.0022 0.0022
70% Perc 0.0025 0.0025 0.0025 0.0025 0.0025
75% Perc 0.0027 0.0027 0.0027 0.0027 0.0027
179
Annexes
Intake of DDT in
Intake of DDT in khat Intake of DDT in khat Intake of DDT in khat Intake of DDT in khat
khat (mg/kg bw/day)
Summary statistics (mg/kg bw/day) Output (mg/kg bw/day) Output (mg/kg bw/day) Output (mg/kg bw/day) Output
Output (Sim#1)
(Sim#2) Sheet1!G8 (Sim#3) Sheet1!G8 (Sim#4) Sheet1!G8 (Sim#5) Sheet1!G8
Sheet1!G8
80% Perc 0.0031 0.0031 0.0031 0.0031 0.0031
85% Perc 0.0035 0.0035 0.0035 0.0035 0.0035
90% Perc 0.0041 0.0041 0.0041 0.0041 0.0041
95% Perc 0.0051 0.0051 0.0051 0.0051 0.0051
Filter Minimum
Filter Maximum
Filter Type
# Values Filtered 0 0 0 0 0
Target #1 (Value) 1
Target #1 (Perc%) 1
Target #2 (Value) 0.0061
Target #2 (Perc%) 0.975
Target #3 (Value) 0.0086
Target #3 (Perc%) 0.995
Target #4 (Value)
Target #4 (Perc%)
180
Annexes
Annex 3: Curriculum Vitae
1. Personal detail
Name: Seblework Mekonen Shegen

Birth date: 11 Sept.1979
Sex: Female
Marital status/number of children: Married/two children
Address: P.O. Box 807, Jimma, Ethiopia.
Email: [email protected], [email protected]
2. Qualification and current position
Assistant professor since June 2014, at Jimma University, Ethiopia
3. Education
From Sept. 2007 – Sept. 2009

Qualification: Master of Science in Environmental Sanitation
Institution: Ghent University, J. Plateaustrat 22, 9000 Ghent, Belgium
From Sept. 2002 - July 2006

Qualification: Bachelor degree in Pharmacy
Institution: Jimma University, P.O. Box 378, Jimma Ethiopia
From Oct. 1997 - Oct. 1999

Qualification: Diploma in Pharmacy
Institution: Jimma Institute of Health Sciences, P.O. Box 378, Jimma Ethiopia
4. Professional carrier
From Sept. 2009 - May 2014

Position: Lecturer, Teaching and research
Major responsibilities: Teaching Bachelor and Master Program students. Main subjects I am
involved are Human Toxicology, Environmental Toxicology, and Environmental Impact of
Pesticides, promoter for under graduate and masters students when doing their thesis.
Name of Employer: Jimma University, P.O. Box 378, Jimma Ethiopia
181
Annexes
From Sept. 2007 – Aug. 2007

Position: Senior Pharmacist, teaching and supervision of students
Major responsibilities: working as senior pharmacist supervising all pharmaceutical cares and
medications in Jimma University Specialized Hospital. I also have been supervising intern
pharmacy students during their practical training in the Hospital.
Name of Employer: Jimma University Specialized Hospital, P.O. Box 378, Jimma, Ethiopia
From Sept. 2001 - Sept. 2002

Position: Druggist, dispensing drugs
Major responsibilities: supervising all pharmaceutical cares and medications in Jimma University
model pharmacy. I also have been supervising intern pharmacy students during their practical
training
Name of Employer: Jimma University Model Pharmacy, P.O. Box 378, Jimma, Ethiopia
From Dec. 1999 - May 2001

Position: Pharmacy Technician
Major responsibilities: Dispensing drugs, giving health information for communities, supervising
private drug stores, venders
Name of Employer: Sagure Health Centre, Arsi Zone Health Bureau, Sagure, Ethiopia
5. Conference presentation: Oral and poster
Oral presentation on the 7th International symposium of Ghent-African Platform-

GAPSYM7, with the theme ‘Africa and Food, Challenges, Risks and Opportunities’. Organized
by Ghent University, December 6.
Oral presentation on the 4th annual conference of Jimma University with the theme of
meeting national development challenges through Science, Technology and Innovations,
February 7-8 and certificate of participation obtained.
Poster presentation on the 65th International Symposium on Crop Protection, organized by
Ghent University, The submitted abstract was published in the proceedings of the
Communication and Agricultural biological Sciences, Vol.78(2), 1-386 (2013). Website:
http://WWW.fbw.ugent.be
Oral presentation on a conference Food and Power, the hunger games in sub-Saharan
African Politics, The commemoration of The World Food Day.Oct.16, 2014.
182
Annexes
Oral presentation on the 26th annual conference of Ethiopian Public Health Association
(EPHA) and certificate of participation obtained
Poster presentation on the 68th international symposium on crop protection, organized by
Ghent University, May 17, 2016
6. Professional affiliation
Member of Ethiopian Pharmaceutical Association (EPA)
Member of Ethiopian Public Health Association (EPHA)
Member of Ethiopian Environmental Health Association (EHA)
7. Short term Training
Basic training on infection prevention and patient safety for higher education teaching staff
organized by USAID, December, 2011.
Training on “Malaria control and Artemisinin based combination Therapy (ACT)

Pharmacovigilance “organized by Drug Administration and Control Authority of Ethiopia
(DACA) in collaboration with Federal Ministry of Health (FMOH) and Ethiopian
Pharmaceutical association (EPA), August, 2007
Pre-service Comprehensive ART training, organized by school of pharmacy Jimma University in

collaboration with Drug Administration and Control Authority of Ethiopia (DACA) and
Management Science for Health (MSH)/RPM Plus-Ethiopia, July, 2006
Training on “Module Writing in Higher Education” organized by Jimma University Academic

Development and Resource Center (ADRC), November, 2009
Training on Business process Re-engineering (BPR), organized by Jimma University, May, 2007
8. List of Publications
Mekonen S, Ambelu A and Spanoghe P, 2014. Pesticide residue evaluation in major staple food
items of Ethiopia using the QuEChERS method: A case study from the Jimma zone.
Environmental Toxicology and Chemistry, Vol. 33, No. 6, pp. 1294–1302.
183
Annexes
Mekonen S. Lachat C., Ambelu A., Steurbaut W., Kolsteren P., Jacxsens L., Wondafrash M.,
Houbraken M. and Spanoghe P.(2015), Risk of DDT residue in maize consumed by infants
as complementary diet in southwest Ethiopia. Science of the Total Environment; Vol.511:
454–460.
Mekonen S, Ambelu A and Spanoghe P, (2015). Effect of Household Coffee Processing on
Pesticide Residues as a Means of Ensuring Consumers’ Safety. Journal of Agricultural and
Food chemistry, DOI: 10.1021/acs.jafc.5b03327
Mekonen S, Argaw R., Simanesew A., Houbraken M., Senaeve D., Ambelu A. and Spanoghe P
(2016). Pesticide residues in drinking water and associated risks to consumers in Ethiopia.
Chemosphere 162 (2016) 252e260, doi.org/10.1016/j.chemosphere.2016.07.096
Mekonen S, Maniales.W.S and Ambelu, 2014. Importance of labeling and patient knowledge to
ensure proper care during drug dispensing: A case study from a tertiary hospital in Ethiopia
Ambelu A, Mekonen S, Koch M, Addis T, Boets P et al, 2014. The application of predictive
modeling for determining Bio-Environmental factors affecting the distribution of Black flies
(Diptera: Simuliidea) in Gilgel Gibe watershed Southwest, Ethiopia. PLos One 9(11),
e112221.doi 10.1371/Journal.Pone.0112221
Getaneh Z, Mekonen S and Ambelu A, 2014. Exposure and health risk assessment of lead in the
communities of Jimma town, Southwest Ethiopia. Bulletin of Environmental contamination
and Toxicology, doi 10.1007/s0028-014-1293-7
Ambelu A, Mekonen S & Gebresilassie A, Malu and Karunamoorthi K.A, 2013.
Physicochemical and Biological characteristics of two Ethiopian wetlands. Wetlands
33:691–698
Amasa W, Santiago D, Mekonen S and Ambelu A, 2012. Are cosmetics used in developing
countries safe? Use and dermal irritation of body care products in Jimma twon, Southwest,
Ethiopia. Journal of Toxicology Volume 2012, Article ID 204830, 8 pages,
doi:10.1155/2012/204830
184

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Human Exposure and Consumer Risk Assessment to Pesticide Use in Ethiopia

Seblework Mekonen Shegen

Prof. dr. ir. Pieter Spanoghe

Department of Crop Protection, Ghent University, Belgium

Dr. Argaw Ambelu

Department of Environmental Health Science and Technology, Jimma University, Ethiopia

Dean: Prof. dr. ir. Marc Van Meirvenne

Rector: Prof. dr. Anne De Paepe

Prof. dr. Ann Dumoulin (Chairman)

Department of Industrial Biological Sciences, Ghent University

Prof. dr. ir. Marijke D’haese (Secretary)

Department of Agricultural Economics, Ghent University

Prof. dr. ir. Patrick Van Damme

Department of Plant Production, Ghent University

Prof. dr. ir. Liesbeth Jacxsens

Department of Food Safety and Food Quality, Ghent University

Prof. dr. Luc Hens

Department of Human Ecology, Vrije Universitiet Brussel

Cover Photo taken and designed by author

Copy right © 2016 Seblework Mekonen Shegen

3.2.4. Analytical equipment .......................................................................................................... 35

7.2.4 Data handling and analysis ................................................................................................... 120

Bestrijdingsmiddelen zijn onmisbaar om de landbouwproductie te verhogen en voor het

Hoofdstuk 3 handelt over de analyse van verschillende bestrijdingsmiddelenresidu’s in het

landbouwkantoren en telers in het studiegebied. De studie omvat ook bestrijdingsmiddelen als

Verschillende studies rapporteren dat de huishoudelijke verwerking van levensmiddelen, zoals

en monitoringprogramma's, de verwerking van levensmiddelen één van de oplossingen zijn om

Naast contaminatie van bovenvermelde conventioneel eetbare voedingsmiddelen en drinkwater,

Om te besluiten, het doctoraatsonderzoek voorziet bewijs van de hoeveelheid contaminatie van

ADI Acceptable Daily Intake

ARfD Acute Reference Dose

CRA Cummulative Risk Assessment

CAG Cumulative Assessment Group

CMG Common Mechanism Group

DDD Dichloro-Diphenyl –Dichloro -Ethane

DDT Dichloro-Diphenyl- Trichloroethane

ECD Electron Capture Detector

EDI Estimated Daily Intake

EFSA European Food Safety Authority

EPA Environmental Protection Agency

EEPA Ethiopian Environmental Protection Authority

ERA Environmental Risk Assessment

ESI Electron Spray Ionization

FAO Food and Agricultural Organization

FFQ Food Frequency Questionnaire

GAP Good Agricultural Practice

GPS Geographical Positioning System

HPLC High Performance Liquid Chromatography

IRS Indoor Residual Spray

ISARC International Sociological Association Research Committee

KOW Octanol-Water Partition Coefficient

LOD Limit of Detection

LOQ Limit of Quantification

LOAEL Lowest Observed Adverse Effect Level

MCRA Monte-Carlo Risk Assessment

MRL Maximum Residue Limit

OCPs Organochlorine Pesticides

POPs Persistent Organic Pollutants

POD Point of Departure