An Exploratory Study of the Metabolite Profiling from Pesticides Exposed Workers.

Pesticides constitute a category of chemical products intended specifically for the control and mitigation of pests. With their constant increase in use, the risk to human health and the environment has increased proportionally due to occupational and environmental exposure to these compounds. The use of these chemicals is associated with several toxic effects related to acute and chronic toxicity, such as infertility, hormonal disorders and cancer. The present work aimed to study the metabolic profile of individuals occupationally exposed to pesticides, using a metabolomics tool to identify potential new biomarkers. Metabolomics analysis was carried out on plasma and urine samples from individuals exposed and non-exposed occupationally, using liquid chromatography coupled with mass spectrometry (UPLC-MS). Non-targeted metabolomics analysis, using principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA) or partial least squares discriminant orthogonal analysis (OPLS-DA), demonstrated good separation of the samples and identified 21 discriminating metabolites in plasma and 17 in urine. The analysis of the ROC curve indicated the compounds with the greatest potential for biomarkers. Comprehensive analysis of the metabolic pathways influenced by exposure to pesticides revealed alterations, mainly in lipid and amino acid metabolism. This study indicates that the use of metabolomics provides important information about complex biological responses.

Metabolomic Study of Urine from Workers Exposed to Low Concentrations of Benzene by UHPLC-ESI-QToF-MS Reveals Potential Biomarkers Associated with Oxidative Stress and Genotoxicity.

Benzene is a human carcinogen whose exposure to concentrations below 1 ppm (3.19 mg·m-3) is associated with myelotoxic effects. The determination of biomarkers such as trans-trans muconic acid (AttM) and S-phenylmercapturic acid (SPMA) show exposure without reflecting the toxic effects of benzene. For this reason, in this study, the urinary metabolome of individuals exposed to low concentrations of benzene was investigated, with the aim of understanding the biological response to exposure to this xenobiotic and identifying metabolites correlated with the toxic effects induced by it. Ultra-efficient liquid chromatography coupled to a quadrupole-time-of-flight mass spectrometer (UHPLC-ESI-Q-ToF-MS) was used to identify metabolites in the urine of environmentally (n = 28) and occupationally exposed (n = 32) to benzene (mean of 22.1 µg·m-3 and 31.8 µg·m-3, respectively). Non-targeted metabolomics analysis by PLS-DA revealed nine urinary metabolites discriminating between groups and statistically correlated with oxidative damage (MDA, thiol) and genetic material (chromosomal aberrations) induced by the hydrocarbon. The analysis of metabolic pathways revealed important alterations in lipid metabolism. These results point to the involvement of alterations in lipid metabolism in the mechanisms of cytotoxic and genotoxic action of benzene. Furthermore, this study proves the potential of metabolomics to provide relevant information to understand the biological response to exposure to xenobiotics and identify early effect biomarkers.

Environmental Assessment and Evaluation of Oxidative Stress and Genotoxicity Biomarkers Related to Chronic Occupational Exposure to Benzene.

Environmental and occupational exposure to benzene from fuels is a major cause for concern for national and international authorities, as benzene is a known carcinogen in humans and there is no safe limit for exposure to carcinogens. The objective of this study was to evaluate the genotoxic effects of chronic occupational exposure to benzene among two groups of workers: filling station workers (Group I) and security guards working at vehicles entrances (Group II), both on the same busy highway in Rio de Janeiro, Brazil. Sociodemographic data on the workers were evaluated; the concentration of benzene/toluene (B/T) in atmospheric air and individual trans,trans-muconic acid (ttMA) and S-phenylmercapturic acid (S-PMA) were measured; oxidative stress was analyzed by catalase (CAT), glutathione S-transferase (GST), superoxide dismutase (SOD), thiol groups (THIOL) and malondialdehyde (MDA); genotoxicity was measured by metaphases with chromosomal abnormalities (MCA) and nuclear abnormalities, comet assay using the enzyme formamidopyrimidine DNA glycosylase (C-FPG), and methylation of repetitive element LINE-1, CDKN2B and KLF6 genes. Eighty-six workers participated: 51 from Group I and 35 from Group II. The B/T ratio was similar for both groups, but Group I had greater oscillation of benzene concentrations because of their work activities. No differences in ttMA and S-PMA, and no clinical changes were found between both groups, but linearity was observed between leukocyte count and ttMA; and 15% of workers had leukocyte counts less than 4.5 × 109 cells L-1, demanding close worker's attention. No differences were observed between the two groups for THIOL, MDA, MCA, or nuclear abnormalities. A multiple linear relationship was obtained for the biomarkers MCA and C-FPG. A significant correlation was found between length of time in current job and the biomarkers C-FPG, MCA, GST, and MDA. Although both populations had chronic exposure to benzene, the filling station workers were exposed to higher concentrations of benzene during their work activities, indicating an increased risk of DNA damage.

A Simple and Quick Method for the Determination of Pesticides in Environmental Water by HF-LPME-GC/MS.

This paper describes a simple and quick method for sampling and also for carrying out the preconcentration of pesticides in environmental water matrices using two-phased hollow fiber liquid phase microextraction (HF-LPME). Factors such as extraction mode, time, solvents, agitation, and salt addition were investigated in order to validate the LPME method. The following conditions were selected: 6 cm of polypropylene hollow fiber, ethyl octanoate as an acceptor phase, and extraction during 30 min under stirring at 200 rpm. The optimized method showed good linearity in the range of 0.14 to 200.00 µg L-1; the determination coefficient (R2) was in the range of 0.9807-0.9990. The LOD ranged from 0.04 µg L-1 to 0.44 µg L-1, and LOQ ranged from 0.14 µg L-1 to 1.69 µg L-1. The recovery ranged from 85.17% to 114.73%. The method was applied to the analyses of pesticides in three environmental water samples (a spring and few streams) collected in a rural area from the state of Minas Gerais, Brazil.

Determination of 2,5-hexanedione in urine by headspace solid-phase microextraction and gas chromatography.

2,5-Hexanedione (2,5-HD) is the most important metabolite of n-hexane and methyl ethyl ketone in human urine. Urinary 2,5-HD is used as a biomarker for biological monitoring of workers exposed to n-hexane. A simple method using headspace solid-phase microextraction (HS-SPME) and gas chromatography (GC) equipped with a flame-ionization detector (FID) was developed. The parameters that affect the HS-SPME-GC-FID process were optimized (i.e., fiber coating, sample volume, adsorption and heating time, salt addition, and extraction temperature). The assay presented linearity in the range of 0.075 to 20.0 mg/L, precision (coefficient of variation < 7.0%), and detection limit of 0.025 mg/L for 2,5-HD in urine. The method was successfully applied to the analysis of 2,5-HD in urine samples from eight workers occupationally exposed to n-hexane in shoemaker's glue.

Hippuric acid in urine: reference values.

OBJECTIVE: To establish reference values for hippuric acid (HA) excreted in the urine, and to evaluate the impact of age, gender, alcohol, and tobacco, on these levels in a population nonexposed to toluene. METHODS: Reference values for hippuric acid in urine were determined in 115 toluene nonexposed healthy volunteers, from Alfenas city, Southeastern Brazil. A questionnaire was applied to each volunteer and data on occupational and personal habits were collected. Biochemical and hematological analyses were used to confirm the volunteers' good health condition. Reference values were expressed in g HA/g urine creatinine, as mean +/- standard deviation (x +/- SD), median, 95% confidence interval (95%CI), 95th percentile, and upper reference value (URV, mean +2 SD). RESULTS: Reference values of hippuric acid in urine were: mean +/- standard deviation =0.18+/-0.10; median =0.15; 95% confidence interval =0.16+/-0.20; 95th percentile = 0.36 and upper reference value (URV, mean +2 SD) =0.38. Statistically significant differences in urinary HA (Wilcoxon - Mann/Whitney, p<0.05) were observed for different genders and age groups. Alcohol ingestion and smoking habit did not significantly affect the results. CONCLUSIONS: The reference values of hippuric acid in urine can be used in biomonitoring programs of workers occupationally exposed to toluene, especially in the southern region of the state of Minas Gerais. Age and gender may affect the HA reference values.

Hippuric acid in urine: reference values

OBJECTIVE: To establish reference values for hippuric acid (HA) excreted in the urine, and to evaluate the impact of age, gender, alcohol, and tobacco, on these levels in a population nonexposed to toluene. METHODS: Reference values for hippuric acid in urine were determined in 115 toluene nonexposed healthy volunteers, from Alfenas city, Southeastern Brazil. A questionnaire was applied to each volunteer and data on occupational and personal habits were collected. Biochemical and hematological analyses were used to confirm the volunteers' good health condition. Reference values were expressed in g HA/g urine creatinine, as mean Ý standard deviation (x Ý SD), median, 95 percent confidence interval (95 percentCI), 95th percentile, and upper reference value (URV, mean +2 SD). RESULTS: Reference values of hippuric acid in urine were: mean Ý standard deviation =0.18Ý0.10; median =0.15; 95 percent confidence interval =0.16Ý0.20; 95th percentile = 0.36 and upper reference value (URV, mean +2 SD) =0.38. Statistically significant differences in urinary HA (Wilcoxon - Mann/Whitney, p<0.05) were observed for different genders and age groups. Alcohol ingestion and smoking habit did not significantly affect the results. CONCLUSIONS: The reference values of hippuric acid in urine can be used in biomonitoring programs of workers occupationally exposed to toluene, especially in the southern region of the state of Minas Gerais. Age and gender may affect the HA reference values