Relationship between Urinary Pesticide Residue Levels and Neurotoxic Symptoms among Women on Farms in the Western Cape, South Africa.

BACKGROUND: This cross-sectional study aimed to investigate the relationship between urinary pesticide residue levels and neurotoxic symptoms amongst women working on Western Cape farms in South Africa. METHOD: A total of 211 women were recruited from farms (n=121) and neighbouring towns (n=90). Participant assessment was via a Q16 questionnaire, reporting on pesticide exposures and measurement of urinary OP metabolite concentrations of dialkyl phosphates (DAP) and chlorpyriphos, 3,5,6-trichloropyridinol (TCPY) and of pyrethroid (PYR) metabolite concentrations (3- phenoxybenzoic acid (3PBA), 4-fluoro-3-phenoxybenzoic acid (4F3PBA), cis-2,2-dibromovinyl-2,2-dimethylcyclopropane-1-carboxylic acid (DBCA), and the cis- and trans isomers of 2,2-dichlorovinyl-2,2-dimethylcyclopropane-1-carboxylic acid. RESULTS: Median urinary pesticide metabolites were slightly (6%-49%) elevated in the farm group compared to the town group, with 2 metabolites significantly higher and some lower in the farm group. The prevalence of all Q16 symptoms was higher amongst farm women compared to town women. Three Q16 symptoms (problems with buttoning, reading and notes) were significantly positively associated with three pyrethroid metabolites (cis- and trans-DCCA and DBCA), although associations may due to chance as multiple comparisons were made. The strongest association for a pyrethroid metabolite was between problems with buttoning and DBCA (odds ratio (OR)=8.93, 95% confidence interval (CI):1.71-46.5. There was no association between Q16 symptoms and OP metabolites. CONCLUSIONS: Women farm residents and rural women from neighbouring towns in the Western Cape are exposed to OP and PYR pesticides. The study did not provide strong evidence that pesticides are associated with neurotoxic symptoms but associations found could be explored further.

Simultaneous monitoring of seven phenolic metabolites of endocrine disrupting compounds (EDC) in human urine using gas chromatography with tandem mass spectrometry.

A gas chromatographic-tandem mass spectrometric (GC-MS/MS) method for the simultaneous determination of the three well-known endocrine disruptors, bisphenol A, daidzein and genistein, as well as of four human pesticide metabolites which are supposed to have proper endocrine activity or which are metabolites of endocrine-disrupting compounds, viz., 1- and 2-naphthol, 2-isopropoxyphenol and 3,5,6-trichloropyridinol, has been developed and validated. The method involves enzymatic cleavage of the conjugates using ß-glucuronidase/arylsulfatase followed by solid-phase extraction and derivatisation with N-tert-butyldimethylsilyl-N-methyltrifluoroacetamide. Isotopically labelled internal standards were used for all analytes, to achieve best analytical error correction. The method proved to be both sensitive and reliable in human urine with detection limits ranging from 0.1 to 0.6 µg/L for all analytes. Precision and repeatability was determined to range from 1 to 15 %. Compared with other published analytical procedures, the present method enables the simultaneous determination of a couple of phenolic agents with competitive or improved analytical reliability. Thus, the present method is suitable for a combined monitoring of the exposure to prominent xenobiotics with effects on the human endocrine system (bisphenol A, carbaryl, chlorpyrifos, chlorpyrifos-methyl, naphthalene, propoxur, triclopyr) and phytoestrogens (daidzein, genistein) in population studies.

Metabolism of 2,6-dichloro-4-(3,3-dichloroallyloxy)phenyl 3-[5-(trifluoromethyl)-2-pyridyloxy]propyl ether (pyridalyl) in rats after repeated oral administration and a simple physiologically based pharmacokinetic modeling in brown and white adipose tissues.

Male and female Sprague-Dawley rats received repeated oral administration of 14C-2,6-dichloro-4-(3,3-dichloroallyloxy)phenyl 3- [5-(trifluoromethyl)-2-pyridyloxy]propyl ether (14C-pyridalyl) at 5 mg/kg/day for 14 consecutive days, and 14C excretion, 14C concentration in tissues, and the metabolic fate were determined. Most 14C was excreted into feces. The 14C concentrations in the blood and tissues attained steady-state levels at days 6 to 10, whereas those in white adipose tissues increased until day 14. Tissue 14C concentrations were highest in brown and white adipose tissue (38.37-57.50 ppm) but were 5.60 ppm or less in all the other tissues. Total 14C residues in blood and tissues on the 27th day after the first administration accounted for 2.6 to 3.2% of the total dose. A major fecal metabolite resulted from O-dealkylation. Analysis of metabolites in tissues revealed that the majority of 14C in perirenal adipose tissue and lungs was pyridalyl, accounting for greater than 90 and 60%, respectively, of the total, whereas a major metabolite in whole blood, kidneys, and liver was a dehalogenated metabolite. The experimental data were simulated with simple physiologically based pharmacokinetics using four-compartment models with assumption of lymphatic absorption and membrane permeability in adipose tissues. The different kinetics in brown and white adipose tissues was reasonably predicted in this model, with large distribution volume in adipose tissues and high hepatic clearance in liver. Sex-related difference of pyridalyl concentration in liver was considered to be a result of different unbound fraction times the hepatic intrinsic clearance (f x CL(int)) of 1.8 and 12 l/h for male and female, respectively.

Metabolism of pyridalyl in rats.

Metabolism of pyridalyl [2,6-dichloro-4-(3,3-dichloroallyloxy)phenyl 3-[5-(trifluoromethyl)-2-pyridyloxy]propyl ether] was examined in male and female Sprague-Dawley rats. After a single oral administration of [dichlorophenyl-(14)C]pyridalyl at 5 or 500 mg/kg, the (14)C concentration in blood reached maxima at 2 to 10 h and then decreased rapidly with a biological half-life of approximately 11 to 12 h. (14)C concentrations in liver, fat, adrenal gland, and spleen were relatively high at a low dose, reaching 2.3 to 2.7, 1.9 to 2.3, 1.1 to 1.9, and 1.4 ppm, respectively, in these tissues at 2 to 24 h after administration. Although (14)C elimination from fat and hair and skin was relatively slow compared with that from other tissues, the total residue on the 7th day was low, in the range of 1.3 to 2.3% of the dose. The (14)C distribution in tissues with a high dose, as examined by whole-body autoradiography, was similar to that observed for the low dose. Results revealed that more than 88% of the dosed radiocarbon was excreted within 1 day after administration, with cumulative (14)C excretion into urine and feces 7 days after administration of 1.7 to 2.6 and 98.7 to 101.7%, respectively. One urinary and fecal major metabolite (resulting from O-dealkylation) and two minor metabolites were identified by NMR and mass spectrometry. Residual (14)C in fat was extracted, and analysis by thin-layer chromatography showed it to be due to pyridalyl itself. No marked sex-related differences were observed in (14)C elimination, (14)C distribution, and metabolites.

Integrated Exposure Assessment Survey (INES) exposure to persistent and bioaccumulative chemicals in Bavaria, Germany.

The Integrated Exposure Assessment Survey (INES) was started in the year 2005. Altogether 50 healthy adults living in Bavaria, Germany, were included into the study. Monitoring was conducted in accordance with relevant routes of human exposure (inhalation, ingestion) and integrated different pathways (indoor air, food, house dust). This approach consisted of a combination of external measurements of contaminants with the determination of these substances or their metabolites in body fluids. The target substances were phthalates, perfluorinated compounds (PFC), polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs), and polybrominated diphenylethers (PBDEs). This paper gives a brief description of the objectives and the concept of INES as well as methods of sampling and analyses of target compounds. Some preliminary results of biomonitoring data for PFC and phthalates as well as of the dietary intake of DEHP will be discussed.

Disposition of 2,2',4,4',5,5'-hexabromodiphenyl ether (BDE153) and its interaction with other polybrominated diphenyl ethers (PBDEs) in rodents.

The disposition of the 14C-labelled polybrominated diphenyl ether (PBDE) 2,2',4,4',5,5'-hexaBDE (BDE153) was investigated in rodents following single and multiple doses and in a mixture with radiolabelled 2,2',4,4'-tetraBDE (BDE47) and 2,2',4,4',5-pentaBDE (BDE99). In single exposure studies there was little or no effect of dose on BDE153 disposition in male rats in the range 1-100 micromol kg-1. No major sex or species differences in the in vivo fate of BDE153 were detected. BDE153 was absorbed in rats or mice following gavage by approximately 70%; retained in tissues; and poorly metabolized and slowly excreted. Mixture studies indicated that, relative to each other, more BDE47 was distributed to adipose tissue, more BDE153 accumulated in the liver, and BDE99 was metabolized to the greatest extent. BDE153 was probably retained in the liver due to minimal metabolism and elimination after 'first-pass' distribution to the tissue following gavage.

Metabolism and disposition of 2,2',4,4',5-pentabromodiphenyl ether (BDE99) following a single or repeated administration to rats or mice.

The metabolism and disposition of 14C-labelled 2,2',4,4',5-pentabromodiphenyl ether (BDE99) were studied in F344 rats and B6C3F1 mice. Approximately 85% of a 1 micromol kg-1 oral dose was absorbed by male rats and mice. Within 24 h following oral doses ranging from 0.1 to 1000 micromol kg-1 to rats, 39-47% of the dose was excreted in the faeces (including 16% unabsorbed), up to 2% was excreted in the urine, and 34-38% remained in the tissues, mostly in adipose tissue. Mice excreted more in the urine and less in the faeces than rats. Tissue accumulation was observed following repeated dosing to rats. Two dihydrohydroxy-S-glutathionyl and two S-glutathionyl conjugates of BDE99, 2,4,5-tribromophenol glucuronide, two mono-hydroxylated BDE99 glucuronides, and three mono-hydroxylated tetrabromodiphenyl ether glucuronides were identified in male rat bile. 2,4,5-Tribromophenol and its glucuronide and sulfate conjugates, were identified in male rat urine. 2,4,5-Tribromophenol, one mono-hydroxylated tetrabromodiphenyl ether, and two mono-hydroxylated BDE99 were characterized in male rat faeces. BDE99 undergoes more extensive metabolism than does BDE47. Half of the absorbed oral dose in male rats was excreted in 10 days mostly as metabolites derived from arene oxide intermediates.

Disposition of BDE 47 in developing mice.

Despite its minor contribution to global polybrominated diphenyl ether (PBDE) production and usage, 2,2',4,4'-tetrabromodiphenyl ether (BDE 47) is the dominant congener found in most biotic samples in North America. The majority of public health concern has focused on potential hazardous effects resulting from exposure of infants and young children to BDE 47 because of previous studies reporting adverse developmental effects in rodent studies, in combination with human exposure estimates suggesting that nursing infants and young children have the highest exposure to BDE 47. This study was designed with two objectives: (1) to investigate the disposition of BDE 47 in infantile mice reported to be susceptible to BDE 47 and (2) to investigate the disposition and excretion of BDE 47 at various developmental stages in an attempt to further identify the mechanism responsible for rapid urinary excretion. The disposition of (14)C-BDE 47 was monitored in C57BL/6 mice following a single oral dose of BDE 47 (1 mg/kg) at different stages of development. The results show that the toxicokinetics of BDE 47 are different in developing mice than in adult mice; whereas disposition patterns are similar, concentrations of BDE 47 are higher in pups because they have a reduced capacity to excrete BDE 47. These differences lead to higher concentrations of BDE 47 at target tissues during critical windows of development.

Impact of repeated exposure on the toxicokinetics of BDE 47 in mice.

2,2',4,4'-Tetrabromodiphenyl ether (BDE 47) is the major polybrominated diphenyl ether (PBDE) found in environmental samples and human tissue despite its small contribution to global production and usage. Currently, three toxicokinetic studies are available investigating single-dose exposures; this is the first study to investigate toxicokinetic parameters following repeated exposure to BDE 47. The disposition and excretion of BDE 47 was monitored in adult female C57BL/6 mice for 5 days following ten consecutive 1.0-mg/kg oral doses and compared with results from our previous study. Results of the present study suggest greater retention of BDE 47 and nonlinear disposition patterns following repeated exposure to this dose in mice. No target tissues of sequestration or potential toxicity were determined; however, some tissues, such as the liver, demonstrated patterns of interest following repeated exposure that were not previously observed in acute toxicokinetic studies. Repeated exposure to BDE 47 results in higher concentrations remaining in adipose tissue, which demonstrates its potential for bioaccumulation. The data also suggest that excretion of BDE 47 may be decreased following repeated exposure. These results, in combination with evidence of its persistence and toxicity, underlie the need to further understand BDE 47 toxicokinetics across species at steady-state conditions.

Gas chromatographic method with mass-selective detection for the determination of 2-isopropoxyphenol in human urine.

Human metabolism of the insecticide propoxur yields 2-isopropoxyphenol (IPP) which is excreted conjugated in urine. In this publication a sensitive and selective analytical method is described which permits the determination of IPP as a suitable parameter for biomonitoring. The clean-up of the hydrolysed urine samples consisted of steam distillation and solid-phase extraction using a reversed-phase column. IPP and the internal standard 2-ethoxyphenol were converted to their pentafluorobenzyl ethers. Excess of the derivatisation reagent was removed using deactivated silica gel. Separation and quantitative analysis was carried out by capillary gas chromatography and mass selective detection. Coefficients of variation were below 5% for concentrations from 6 to 300 microg/l. The detection limit was 0.5 microg/l. The method was checked by analysing six urine samples from pest controllers after indoor application of propoxur. The IPP concentrations ranged from 45 to 306 microg/g creatinine. IPP was not detected in urine specimens from 10 non-exposed persons. The sensitivity of the developed method permits the detection of latent exposure to propoxur.

[The use of biological monitoring in the assessment of occupational exposure to styrene in fiberglass-reinforced plastics industry]. / Zastosowanie monitoringu biologicznego do oceny zawodowego narazenia na styren w przemysle tworzyw sztucznych.

A field study was carried out in order to investigate the effect of occupational exposure to styrene on the urinary excretion of mandelic and phenylglyoxylic acids (MA and PGA). Over a period of three years, 210 periodical, prophylactic examinations were performed on 66 fiberglass-reinforced plastics workers. The examinations embraced environmental and biological monitoring, which involved the measurement of styrene concentration in the air and the excretion rate of urinary MA and PGA. An eight-hour time-weighed average (MAC) exposure values ranged from 16.1 to 246 mg/m3 (mean: 76.1 mg/m3). The average urinary excretion rates of MA and PGA were 17.7 (1-202) and 28.8 (2-267) mg/h, respectively. The measurements of MA excretion rate were compared with the measurements of styrene concentration in the air. It seems advisable to investigate whether the recommended biological limits value of 16 mg/h for mandelic acid is not too high in view of the binding OEL value of 50 mg/m3 for styrene.

The influence of skin moisture on the dermal absorption of propoxur in human volunteers: a consideration for biological monitoring practices.

A large number of workers in agriculture are exposed daily (through skin contact) to pesticides either directly during mixing and loading or indirectly due to contact. The aim of this study was to investigate the influence of skin moisture on the dermal uptake of the pesticide propoxur. The study was conducted in human volunteers under controlled temperature conditions (30 degrees C) and environmental relative humidities of either 50, 70 or 90%. The study was approved by the Medical Ethics Committee. In this study a linear relationship between the environmental relative humidity and the level of skin moisture was observed. The results indicate that the level of skin moisture influences the absorption of propoxur via the dermal route, dramatically ranging from, on average, 13, 33-63% of the potentially absorbed dose' which is excreted in urine as the primary metabolite 2-isopropoxyphenol (IPP) at relative humidity levels of, on average 50, 70 and 90%, respectively. The 'potentially absorbed dose' is defined as the difference between the applied dose and the dislodged dose after 4 h. It can be concluded that by assessing health risks of workers in agriculture exposed dermally to pesticides and e.g. in testing the efficiency of protective clothing under realistic conditions, the influence of the level of skin moisture on absorption of substances may be considerable and has to be taken into account.

Use of fresh and cryopreserved hepatocytes to study the metabolism of pesticides in food-producing animals and rats.

1. The possibility of using hepatocytes from food-producing animals in order to determine the metabolic routes of pesticides has been studied using a strobilurin fungicide (BAS 490 F). Hepatocytes suspensions were prepared from goat, pig, hen, and rat and the major metabolites were compared with those obtained in vivo. 2. The hepatocytes gave metabolite patterns matching qualitatively with in vivo results, but no good quantitative correlation was found. 3. A freezing and thawing method was developed using liquid nitrogen and a programmable freezer, which allows acceptable recoveries of functional cells as assessed by glutathione and cytochrome P450 contents, and phase I and II enzymatic activities (including 7-ethoxycoumarin-O-deethylase, ethoxyresorufin-O-deethylase, glutathione-S-transferase, and UDP-glucuronosyl transferase), with 60-70% viability. 4. The cells were damaged through freezing as indicated by the efflux of glutathione (40-60% of the intracellular content), but remained able to metabolize BAS 490 F, partially like fresh cells. A good qualitative but no quantitative matching of the metabolite patterns before and after cryopreservation was found, indicating that the metabolic activities are affected to variable extents during the freezing process. 5. The use of fresh and cryopreserved cells as models for metabolism and species comparison, and as a versatile tool to synthesize metabolites, is discussed.

Skin contamination, airborne concentrations, and urinary metabolite excretion of propoxur during harvesting of flowers in greenhouses.

In eight greenhouses used for carnation culture, workers engaged in harvesting (n = 16), were monitored for dermal and respiratory exposure and urinary excretion of propoxur. Dermal exposure of hands and forearms was estimated from dislodgable foliar residue, using a transfer factor (a measure of transfer of pesticides from leaves to the skin) and the total number of working hours. Total estimated dermal and respiratory exposure during harvesting ranged from 0.2 to 46 mg and from 3 to 278 micrograms, respectively. To study the relationship between external and internal exposure to propoxur, respiratory and dermal exposure levels were compared with the total amount of 2-isopropoxyphenol (IPP), the major metabolite of propoxur, excreted in urine in 24 hr. The Pearson correlation coefficient between dermal exposure and the total amount of excreted IPP was 0.95. A correlation coefficient of 0.84 was found between respiratory exposure and the amount of IPP excreted. The latter association was probably caused by the covariation of respiratory and dermal exposure levels (r = 0.85). Assuming negligible oral absorption, calculations indicated that dermal exposure could account for > 80% of the amount of excreted IPP. On the basis of the amount of IPP excreted, there was no reason to suspect increased health risks for workers from exposure to propoxur during harvesting.

Determination of 2-isopropoxyphenol in urine by capillary gas chromatography and mass-selective detection.

An analytical method for the assessment of the exposure of workers to the pesticide propoxur through biological monitoring has been developed. This study was part of a survey of occupational exposure to pesticides used in greenhouses for the growing of ornamental plants. In order to assess the actual absorbed amount of propoxur in the body, an analytical method for its metabolite 2-isopropoxyphenol in urine was required. This led to the development of a gas chromatographic-mass spectrometric assay involving hydrolysis and solvent extraction. A mass-selective detector, operated in single-ion mode, provides a selective and sensitive quantification of 2-isopropoxyphenol with a detection limit of 6 micrograms/l. The method has been validated with respect to the hydrolysis of urine samples, analytical recovery of 2-isopropoxyphenol, stability of its conjugates, limit of detection, background and precision. The analytical recovery from spiked urine was over 95%. 2-Isopropoxyphenol was excreted in urine as a conjugate and was stable for at least seven months when stored at -20 degrees C. It was not detected in urine from non-exposed persons. Between-day coefficients of variation were 20, 10, 7 and 4% for concentrations of 15, 29, 150 and 213 micrograms/l, respectively. Measured as 2-isopropoxyphenol, ca. 80% of an orally administered dose of propoxur was excreted in urine within 10 h.

Determination of the herbicide diclofop in human urine.

A simple and sensitive method for the gas chromatographic determination of diclofop residues in human urine is described. Recoveries of diclofop, as its methyl ester, from fortified urine were greater than 85% at 100, 50, 10 and 1 micrograms kg-1, and were similar with and without the inclusion of a hydrolytic step in the analytical method. However, a hydrolytic step was necessary for analysis of 24-h urine samples collected from a male applicator following a single exposure to diclofop-methyl during application to wheat using a tractor-pulled sprayer. Diclofop residues determined with hydrolysis were approximately double those without hydrolysis, suggesting that a significant portion of diclofop was excreted in the conjugated form.

Biotransformation of diphenyl ether by trout and guinea-pigs after intraperitoneal administration.

Diphenyl ether (DPE) was administered intraperitoneally to rainbow trout (Salmo gairdneri) and guinea-pigs (Duncan Hartley). DPE metabolites in the urine and bile of trout and the urine of guinea-pigs were isolated and analysed by g.l.c.-mass spectrometry. No unchanged DPE was found in the biological fluids of guinea-pigs and trout. Only conjugated metabolites of DPE were present in the bile and urine of the trout: 4-hydroxy- and possible 3-hydroxy-DPE conjugates were found in the bile whereas 4-hydroxy- and 4,4-dihydroxy-DPE conjugates were detected in the urine. Both free and conjugated metabolites of DPE were isolated from the urine of guinea-pigs. These metabolites were 2-hydroxy-, 4-hydroxy, 4,4'-dihydroxy-, 4-methoxy-monohydroxy- and methoxy-dihydroxy-derivatives of DPE.

The metabolism of 3-phenoxybenzoic acid and its glucoside conjugate in rats.

1. 3-Phenoxy[14C]benzoic acid administered orally to rats (0.76 to > 100 mg/kg) is extensively metabolized and rapidly eliminated mostly via the urine. 2. The major metabolic pathway involves 4'-hydroxylation followed by conjugation of the resulting phenol with sulphate. Only minor amounts of amino acid and glucuronic acid conjugation were observed. 3. 3-Phenoxy[14C]benzoyl glucoside, derived from the metabolism of the acid by corn leaves, was also rapidly absorbed by rats and eliminated as a mixture of metabolites very similar to that derived from 3-phenoxybenzoic acid. It was concluded that the glucoside conjugate was rapidly hydrolysed to the free acid in vivo.

Taurine conjugation in metabolism of 3-phenoxybenzoic acid and the pyrethroid insecticide cypermethrin in mouse.

1. A substantial proportion of the radioactivity of an oral dose (1-20 mg/kg) of 3-phenoxy [14C] benzoic acid and the pyrethroid insecticide trans- and cis-[aryl-14C] cypermethrin (an alpha-cyano-3-phenoxybenzyl ester) is eliminated rapidly in urine of mice. 2. The major urinary metabolite of these compounds in two strains of mice is N-(3-phenoxybenzoyl)taurine. 3. Neither 3-phenoxybenzoic acid nor cypermethrin gives a taurine conjugate in rats. 4. Benzoic acid gives no taurine conjugate in mice.