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Two years of a biomonitoring study of mercury in human hair and urine, Madrid (Spain)
Abstracts / Toxicology Letters 196S (2010) S37–S351
P101-007 Controlled human exposures to organic solvents: Influence of sex on urinary levels of biological indicators C. Tomicic a , P.O. Droz b , M. Berode b , S.M. Praz Christinaz b , V. Castella c , B. Danuser b Federal Office of Public Health (FOPH), Switzerland, b Institute for Work and Health (IST), Switzerland, c Unité de Génétique Forensique, Centre Universitaire Romand de Médecine Légale, Switzerland
a
Human biomonitoring is a method widely used in the assessment of occupational exposure to chemical substances. Recommended biological limits are published periodically for interpretation and decision-making. However it is increasingly recognized that a large variability is associated with biological monitoring, making interpretation less efficient than assumed. Among others, sex is an easily identifiable determinant which could play an important contribution to biological variability. The aim of the study was to quantify the variability on biological indicators between men and women for three well-known solvents: methyl ethyl ketone, 1-methoxy-2-propanol and 1,1,1trichloroethane. Controlled human exposures were carried out in a 12 m3 exposure chamber for each solvent separately, during 6 h and at half of the threshold limit value. The human volunteers groups were composed of 10 young men and 15 young women. Analyses of the solvent and their metabolites were performed by using headspace gas chromatography, CYP2E1 genotypes by using PCR-based RFLP methods. The results obtained for the different biomarkers of exposure indicate a smaller interindividual variability among men as among women. An analysis of variance showed following main observations: - a negative correlation between body fat and urinary 1-methoxy2-propanol concentrations; - a statistical significant difference due to the genotype CYP2E1*6 with a tendency to increase CYP2E1 activity when the volunteers are exposed to methyl ethyl ketone; - an effect among women due to the use of hormonal contraceptive on the urinary levels of several biomarkers with an increase of 50% in the metabolism rate. Combination oral contraceptives increase estrogen and progesterone levels and decrease gonadotropins level. Our study suggests that exogenous sex hormones could influence CYP2E1 enzyme activity. The variability among urinary biological indicators levels gives evidence of an interindividual susceptibility, an aspect that should have its place in the approaches for setting limits of occupational exposure.
S39
extraction. Isotretinoin and internal standard, acitretine, were separated on Eclipse PLUS C18 column (4.6 mm × 150 mm, 3.5 m) with a mobile phase. The mass transitions m/z 299.2 → 255.3 and m/z 299.2 → 255.3 were used to measure isotretinoin and acitretine, respectively. Calibration curve was linear in the concentration range of 0.1–30 ng/ml for isotretinoin in human serum. For quality control samples at three different concentrations that were analyzed in independent five experiments, the accuracy and precision range from 92% to 103.9% and 3.9% to 7.5%, respectively. The validated method was used to analyze isotretinoin in 204 human serums. doi:10.1016/j.toxlet.2010.03.167
P101-009 Human biomonitoring of phthalates using LC/MS/MS in South Korea H.S. Nam, S.K. Hong, I.H. Kang, T.S. Kim, K.K. Jung, H.J. Yoon, T.S. Kang Korea Food and Drug Administration Phthalates are ubiquitous toxic chemicals widely used in consumer products as solvents, additives, cosmetics, and plasticizers. Some phthalates are known to cause carcinogenic or reproductive toxicities in laboratory animals. A study on the exposure assessment of phthalates is needed because human can be exposed to phthalates through ingestion, inhalation, dermal absorption, by using phthalate containing products. In this study, MnBP, MiBP, MBzP, MCHP, MEHP, MEHHP, MEOHP, MnOP, MiNP, and MiDP, which are monoetster metabolites of DEHP, DBP, BBP, DNOP, DIDP, DCHP, were chosen and analyzed in urine samples from the South Korean population. We developed a simultaneous analysis method for 10 phthalate metabolites, based on switching system with on-line pretreatment column using LC/MS/MS. We estimated urinary creatinine corrected geometric means of 10 phthalate metabolites in 497 general populations, recruited in Chounchenog-do, Jeolla-do, and Gangwon-do (women, n = 273, and men, n = 224). Eight phthalate metabolites, except for MnOP and MiDP, were detected in most of urine samples. The results of statistical analysis according to sex, age and region show that women trend to have greater level in most of phthalates. We confirmed that daily exposure levels of DEHP, DBP and BBP were less than the tolerable daily intake (TDI) of the KFDA. doi:10.1016/j.toxlet.2010.03.168
P101-010 Two years of a biomonitoring study of mercury in human hair and urine, Madrid (Spain)
doi:10.1016/j.toxlet.2010.03.166 ˜ M. Esteban, A. Canas, C. Navarro, J.E. Sánchez, J.A. Jiménez, M. ˜ Arribas, J.P. García, A. Castano P101-008 Determination of isotretinoin in human serum by LC–MS/MS H.S. Nam, H.J. Yoon, K.K. Jung, I.H. Kang, T.S. Kim, S.K. Hong, T.S. Kang Korea Food and Drug Administration, Republic of Korea The purpose of this study is to develop and validate for quantitative determination of isotretinoin in human serum using LC–MS/MS. The pretreatment of samples involves solid-phase
Instituto de Salud Carlos III Mercury is a well-known toxicant and is ranked in the first positions on the ATSDR/EPA priority hazardous substances. The most reliable way to measure mercury exposure due to fish consumption and other sources for general population such as amalgam fillings is by human biomonitoring. In the last years the Environmental Toxicology Unit of ISCIII has been conducting a pilot study based on measurements of chemicals in non-invasive human matrices including mercury and
S40
Abstracts / Toxicology Letters 196S (2010) S37–S351
methyl mercury in scalp hair and mercury in urine. Total mercury in scalp hair (H-THg) was determined by AA spectrometry and same was used for methyl mercury (H-MeHg) with a previous toluene/N-acetyl l-cysteine acid extraction. Urinary mercury (UHg) was measured by ICP-MS. The study population was formed by workers of the ISCIII that went to the annual medical revision and agreed to donate a first-morning urine sample, a hair sample and completed a questionnaire. The population studied was recruited in two consecutive surveys (2007 and 2008) involving approximately 260 volunteers each one, 79% were female and 21% male, and between 21 – 69 years old. H-THg levels ranged from 0.11 to 15.97 g/g and H-MeHg ranged from
P101-011 Markers of oxidative stress and genotoxicity applied in rural workers exposed to pesticides M.F. Simoniello 1 , E.C. Kleinsorge 1 , M.A. Carballo 2 Universidad Nacional del Litoral, 2 CIGETOX-Citogenética Humana y Genética Toxicológica, INFIBIOC, UBA
1
Pesticides are used in agriculture to protect crops but may represent a potential risk to farmers and environment. In spite of the economic advantages, in our country there is no control on the applicators working conditions, including general protective measures. Occupational exposure occurs during the preparation of mixtures, loading and/or washing equipment and spraying on crops. In addition, individuals are often exposed to different pesticides or pesticide mixtures, either simultaneously or in series, making it difficult to identify the effects of each one separately. In this frame, exposure and effect biomarkers can be used to detect alterations induced by pesticides in human tissues, occurring before the appearance of adverse health effects. We report data obtained from a wide evaluation of workers, directly or indirectly exposed to pesticides (applicators and farmers) and control group, including oxidative balance, exposure biomarkers and DNA damage quantification. Selected methodologies were: Cholinesterase (ChE) and Acetylcholinesterase (AChE) activities, Catalase (CAT), Lipid Peroxidation (by TBARS assay), Damage Index Comet Assay (IDEC) and Damage Index Repair Assay (IDER). The results shows: a) AChE inhibition in directly and indirectly exposed population, b) significant increase in the levels of TBARS was found in pesticide sprayers, c) CAT reduction was significant in both groups, d) significant increase in IDEC and IDER was found in both exposed groups. Our results evidence variations in oxidative balance as well as lipid peroxidation and DNA damage in directly and indirectly exposed subjects. It has to be pointed out that Multivariate analysis demonstrated a significant correlation between variables but not an influence of confounding factors on the genotoxic effects
of occupational exposure to pesticides. These findings represent a contribution in sub-clinical evaluation of subjects occupationally exposed to potential genotoxic agrochemicals in our country. doi:10.1016/j.toxlet.2010.03.170
P101-012 Reference values for lead, cadmium and mercury in blood of adults from the metropolitan area of São Paulo (Brazil) R. Kuno 1 , M.H. Roquetti 1 , K. Becker 2 , M. Seiwert 2 , N. Gouveia 3 CETESB - Companhia Ambiental do Estado de São Paulo, 2 German Federal Environmental Agency, 3 Faculdade de Medicina, Universidade de São Paulo
1
The aim of this study was to derive reference values (RV), that is the 95th percentile of the background exposure, for lead, cadmium and mercury in blood of adults from the Metropolitan Area of Sao Paulo (Brazil), and to investigate the association between blood metals and sociodemographic and lifestyle factors. Blood samples of 653 blood donors, non-smokers and without any occupational exposure to the studied metals were collected in 2006. In our evaluations we distinguished between the younger group (18–39 years) and the older group (40–65 years). RV derived for the younger group were for men 59.73 g/L for Pb, 0.41 g/L for Cd and 4.30 g/L for Hg; for women 47.09 g/L for Pb, 0.48 g/L for Cd and 3.71 g/L for Hg. For the older group were for men 79.84 g/L for Pb, 0.35 g/L for Cd and 5.10 g/L for Hg; for women 63.10 g/L for Pb, 0.44 g/L for Cd and 6.10 g/L for Hg. The RV for lead were similar to the Czech Republic and Germany, but higher than those observed in a study of the U.S. population (NHANES). RV for Cd were well below of those from these countries. The RV for Hg were similar to the U.S. and higher than the value in Germany and in the Czech Republic. Pb and Cd in blood showed a significant association with sex and age. Men had 50% more lead in blood, and the older group showed 23% more lead than those in 18–39 years old. The variables most related to the levels of Hg were fish consumption and amalgam fillings. Also, the older group had 19% more Hg than the younger group. Individuals with basic education showed significantly lower mercury levels than those with higher education. doi:10.1016/j.toxlet.2010.03.171
P101-013 Improved procedure for biomonitoring 28 elements in human hair: Sample collection, treatment and analytical method by ICP-MS E. Vilanova, R. Ruiz, C. Estevan, M.A. Sogorb Universidad Miguel Hernández, Spain The full procedure was optimized for collecting of human hair samples, treatment of samples and measuring by ICP-MS for biomonitoring 28 elements (Be-B-Na-Mg-Al-K-Ca-V-Cr-Mn-Fe-Co-Ni-CuZn-As-Se-Sr-Mo-Cd-Ba-Hg-Tl-Pb-Bi-Ag-Au-U) with interest either as environmental pollutants or as oligoelements of nutritional interest. Human hair sample are easy to collect, store and transport. Application in surveys to general population is possible as noninvasive samples. Several strategies were tested and improved. Sampling was facilitated by informative actions, a simplified graphic instruction and a questionnaire for getting the substantial information of variables (age, sex, geographical place, and nutritional habits and chronic diseases). Sample of about 200 mg hair
P101-007 Controlled human exposures to organic solvents: Influence of sex on urinary levels of biological indicators C. Tomicic a , P.O. Droz b , M. Berode b , S.M. Praz Christinaz b , V. Castella c , B. Danuser b Federal Office of Public Health (FOPH), Switzerland, b Institute for Work and Health (IST), Switzerland, c Unité de Génétique Forensique, Centre Universitaire Romand de Médecine Légale, Switzerland
a
Human biomonitoring is a method widely used in the assessment of occupational exposure to chemical substances. Recommended biological limits are published periodically for interpretation and decision-making. However it is increasingly recognized that a large variability is associated with biological monitoring, making interpretation less efficient than assumed. Among others, sex is an easily identifiable determinant which could play an important contribution to biological variability. The aim of the study was to quantify the variability on biological indicators between men and women for three well-known solvents: methyl ethyl ketone, 1-methoxy-2-propanol and 1,1,1trichloroethane. Controlled human exposures were carried out in a 12 m3 exposure chamber for each solvent separately, during 6 h and at half of the threshold limit value. The human volunteers groups were composed of 10 young men and 15 young women. Analyses of the solvent and their metabolites were performed by using headspace gas chromatography, CYP2E1 genotypes by using PCR-based RFLP methods. The results obtained for the different biomarkers of exposure indicate a smaller interindividual variability among men as among women. An analysis of variance showed following main observations: - a negative correlation between body fat and urinary 1-methoxy2-propanol concentrations; - a statistical significant difference due to the genotype CYP2E1*6 with a tendency to increase CYP2E1 activity when the volunteers are exposed to methyl ethyl ketone; - an effect among women due to the use of hormonal contraceptive on the urinary levels of several biomarkers with an increase of 50% in the metabolism rate. Combination oral contraceptives increase estrogen and progesterone levels and decrease gonadotropins level. Our study suggests that exogenous sex hormones could influence CYP2E1 enzyme activity. The variability among urinary biological indicators levels gives evidence of an interindividual susceptibility, an aspect that should have its place in the approaches for setting limits of occupational exposure.
S39
extraction. Isotretinoin and internal standard, acitretine, were separated on Eclipse PLUS C18 column (4.6 mm × 150 mm, 3.5 m) with a mobile phase. The mass transitions m/z 299.2 → 255.3 and m/z 299.2 → 255.3 were used to measure isotretinoin and acitretine, respectively. Calibration curve was linear in the concentration range of 0.1–30 ng/ml for isotretinoin in human serum. For quality control samples at three different concentrations that were analyzed in independent five experiments, the accuracy and precision range from 92% to 103.9% and 3.9% to 7.5%, respectively. The validated method was used to analyze isotretinoin in 204 human serums. doi:10.1016/j.toxlet.2010.03.167
P101-009 Human biomonitoring of phthalates using LC/MS/MS in South Korea H.S. Nam, S.K. Hong, I.H. Kang, T.S. Kim, K.K. Jung, H.J. Yoon, T.S. Kang Korea Food and Drug Administration Phthalates are ubiquitous toxic chemicals widely used in consumer products as solvents, additives, cosmetics, and plasticizers. Some phthalates are known to cause carcinogenic or reproductive toxicities in laboratory animals. A study on the exposure assessment of phthalates is needed because human can be exposed to phthalates through ingestion, inhalation, dermal absorption, by using phthalate containing products. In this study, MnBP, MiBP, MBzP, MCHP, MEHP, MEHHP, MEOHP, MnOP, MiNP, and MiDP, which are monoetster metabolites of DEHP, DBP, BBP, DNOP, DIDP, DCHP, were chosen and analyzed in urine samples from the South Korean population. We developed a simultaneous analysis method for 10 phthalate metabolites, based on switching system with on-line pretreatment column using LC/MS/MS. We estimated urinary creatinine corrected geometric means of 10 phthalate metabolites in 497 general populations, recruited in Chounchenog-do, Jeolla-do, and Gangwon-do (women, n = 273, and men, n = 224). Eight phthalate metabolites, except for MnOP and MiDP, were detected in most of urine samples. The results of statistical analysis according to sex, age and region show that women trend to have greater level in most of phthalates. We confirmed that daily exposure levels of DEHP, DBP and BBP were less than the tolerable daily intake (TDI) of the KFDA. doi:10.1016/j.toxlet.2010.03.168
P101-010 Two years of a biomonitoring study of mercury in human hair and urine, Madrid (Spain)
doi:10.1016/j.toxlet.2010.03.166 ˜ M. Esteban, A. Canas, C. Navarro, J.E. Sánchez, J.A. Jiménez, M. ˜ Arribas, J.P. García, A. Castano P101-008 Determination of isotretinoin in human serum by LC–MS/MS H.S. Nam, H.J. Yoon, K.K. Jung, I.H. Kang, T.S. Kim, S.K. Hong, T.S. Kang Korea Food and Drug Administration, Republic of Korea The purpose of this study is to develop and validate for quantitative determination of isotretinoin in human serum using LC–MS/MS. The pretreatment of samples involves solid-phase
Instituto de Salud Carlos III Mercury is a well-known toxicant and is ranked in the first positions on the ATSDR/EPA priority hazardous substances. The most reliable way to measure mercury exposure due to fish consumption and other sources for general population such as amalgam fillings is by human biomonitoring. In the last years the Environmental Toxicology Unit of ISCIII has been conducting a pilot study based on measurements of chemicals in non-invasive human matrices including mercury and
S40
Abstracts / Toxicology Letters 196S (2010) S37–S351
methyl mercury in scalp hair and mercury in urine. Total mercury in scalp hair (H-THg) was determined by AA spectrometry and same was used for methyl mercury (H-MeHg) with a previous toluene/N-acetyl l-cysteine acid extraction. Urinary mercury (UHg) was measured by ICP-MS. The study population was formed by workers of the ISCIII that went to the annual medical revision and agreed to donate a first-morning urine sample, a hair sample and completed a questionnaire. The population studied was recruited in two consecutive surveys (2007 and 2008) involving approximately 260 volunteers each one, 79% were female and 21% male, and between 21 – 69 years old. H-THg levels ranged from 0.11 to 15.97 g/g and H-MeHg ranged from
P101-011 Markers of oxidative stress and genotoxicity applied in rural workers exposed to pesticides M.F. Simoniello 1 , E.C. Kleinsorge 1 , M.A. Carballo 2 Universidad Nacional del Litoral, 2 CIGETOX-Citogenética Humana y Genética Toxicológica, INFIBIOC, UBA
1
Pesticides are used in agriculture to protect crops but may represent a potential risk to farmers and environment. In spite of the economic advantages, in our country there is no control on the applicators working conditions, including general protective measures. Occupational exposure occurs during the preparation of mixtures, loading and/or washing equipment and spraying on crops. In addition, individuals are often exposed to different pesticides or pesticide mixtures, either simultaneously or in series, making it difficult to identify the effects of each one separately. In this frame, exposure and effect biomarkers can be used to detect alterations induced by pesticides in human tissues, occurring before the appearance of adverse health effects. We report data obtained from a wide evaluation of workers, directly or indirectly exposed to pesticides (applicators and farmers) and control group, including oxidative balance, exposure biomarkers and DNA damage quantification. Selected methodologies were: Cholinesterase (ChE) and Acetylcholinesterase (AChE) activities, Catalase (CAT), Lipid Peroxidation (by TBARS assay), Damage Index Comet Assay (IDEC) and Damage Index Repair Assay (IDER). The results shows: a) AChE inhibition in directly and indirectly exposed population, b) significant increase in the levels of TBARS was found in pesticide sprayers, c) CAT reduction was significant in both groups, d) significant increase in IDEC and IDER was found in both exposed groups. Our results evidence variations in oxidative balance as well as lipid peroxidation and DNA damage in directly and indirectly exposed subjects. It has to be pointed out that Multivariate analysis demonstrated a significant correlation between variables but not an influence of confounding factors on the genotoxic effects
of occupational exposure to pesticides. These findings represent a contribution in sub-clinical evaluation of subjects occupationally exposed to potential genotoxic agrochemicals in our country. doi:10.1016/j.toxlet.2010.03.170
P101-012 Reference values for lead, cadmium and mercury in blood of adults from the metropolitan area of São Paulo (Brazil) R. Kuno 1 , M.H. Roquetti 1 , K. Becker 2 , M. Seiwert 2 , N. Gouveia 3 CETESB - Companhia Ambiental do Estado de São Paulo, 2 German Federal Environmental Agency, 3 Faculdade de Medicina, Universidade de São Paulo
1
The aim of this study was to derive reference values (RV), that is the 95th percentile of the background exposure, for lead, cadmium and mercury in blood of adults from the Metropolitan Area of Sao Paulo (Brazil), and to investigate the association between blood metals and sociodemographic and lifestyle factors. Blood samples of 653 blood donors, non-smokers and without any occupational exposure to the studied metals were collected in 2006. In our evaluations we distinguished between the younger group (18–39 years) and the older group (40–65 years). RV derived for the younger group were for men 59.73 g/L for Pb, 0.41 g/L for Cd and 4.30 g/L for Hg; for women 47.09 g/L for Pb, 0.48 g/L for Cd and 3.71 g/L for Hg. For the older group were for men 79.84 g/L for Pb, 0.35 g/L for Cd and 5.10 g/L for Hg; for women 63.10 g/L for Pb, 0.44 g/L for Cd and 6.10 g/L for Hg. The RV for lead were similar to the Czech Republic and Germany, but higher than those observed in a study of the U.S. population (NHANES). RV for Cd were well below of those from these countries. The RV for Hg were similar to the U.S. and higher than the value in Germany and in the Czech Republic. Pb and Cd in blood showed a significant association with sex and age. Men had 50% more lead in blood, and the older group showed 23% more lead than those in 18–39 years old. The variables most related to the levels of Hg were fish consumption and amalgam fillings. Also, the older group had 19% more Hg than the younger group. Individuals with basic education showed significantly lower mercury levels than those with higher education. doi:10.1016/j.toxlet.2010.03.171
P101-013 Improved procedure for biomonitoring 28 elements in human hair: Sample collection, treatment and analytical method by ICP-MS E. Vilanova, R. Ruiz, C. Estevan, M.A. Sogorb Universidad Miguel Hernández, Spain The full procedure was optimized for collecting of human hair samples, treatment of samples and measuring by ICP-MS for biomonitoring 28 elements (Be-B-Na-Mg-Al-K-Ca-V-Cr-Mn-Fe-Co-Ni-CuZn-As-Se-Sr-Mo-Cd-Ba-Hg-Tl-Pb-Bi-Ag-Au-U) with interest either as environmental pollutants or as oligoelements of nutritional interest. Human hair sample are easy to collect, store and transport. Application in surveys to general population is possible as noninvasive samples. Several strategies were tested and improved. Sampling was facilitated by informative actions, a simplified graphic instruction and a questionnaire for getting the substantial information of variables (age, sex, geographical place, and nutritional habits and chronic diseases). Sample of about 200 mg hair