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Background Serious concerns surround the potential risks resulting from inhalation exposure to pesticides amongst agricultural workers when mixing and applying these compounds. In Eswatini (formerly known as Swaziland), Southern Africa, pesticides are widely used to improve the yield and quality of sugar cane production, the largest contributor to the country's economy. We assessed applicators' inhalation exposures from multiple spraying sources to four commonly used herbicides in Eswatini. Results Analysis of 76 personal air samples by liquid chromatography with tandem mass spectrometry (LC-MS/MS) revealed four pesticides: ametryn, atrazine, pendimethalin and 2,4-dichlorophenoxyacetic acid, with mean concentrations of 36.91, 21.57, 31.05 and 0.89 μg m-3 , respectively. These inhalation exposures are much higher than those recorded in previous similar studies. Conclusion Although all applicators in this study used personal protective equipment (PPE), they nevertheless recorded high levels of inhalation exposure to commonly used pesticides. Our findings suggest that in addition to observing mandated regular changing and cleaning practices with PPE for ultimate personal protection, pesticide applicators should distance themselves from each other when spraying to effectively reduce their exposure to pesticides from multiple spraying sources. Further studies are needed to determine the optimal spraying distance between pesticide applicators. © 2021 Society of Chemical Industry.

Exposure to the vinyl monomer acrylonitrile (AN) is primarily occupational. AN is also found in cigarette smoke. AN can be detoxified to form N-acetyl-S-(2-cyanoethyl)-cysteine (CEMA) or activated to 2-cyanoethylene oxide (CEO) and detoxified to form N-acetyl-S-(1-cyano-2-hydroxyethyl)-cysteine (CHEMA) and N-acetyl-S-(2-hydroxyethyl)-cysteine (HEMA). These urinary mercapturic acids (MAs) are considered to be potential biomarkers of AN exposure. This study assessed personal AN exposure, urinary MAs (CEMA, CHEMA, and HEMA), and cotinine (a biomarker of cigarette smoke) in 80 AN-exposed and 23 non-exposed factory workers from urine samples provided before and after work shifts. Unambiguous linear correlations were observed between levels of urinary CEMA and CHEMA with personal AN exposures, indicating their potential as chemically-specific biomarkers for AN exposures. AN exposure was the dominant factor in MA formation for AN-exposed workers, whereas urinary cotinine used as a biomarker showed that cigarette smoke exposure was the primary factor for non-exposed workers. The CHEMA/CEMA and (CHEMA+HEMA)/CEMA ratios in this human study differ from those in similar studies of AN-treated rats and mice in literature, suggesting a possible dose- and species-dependent effect in AN metabolic activation and detoxification.

Background Epidemiological evidence has linked fine particulate matter (PM2.5) to neurodegenerative diseases; however, the toxicological evidence remains unclear. The objective of this study was to investigate the effects of PM2.5 on neuropathophysiology in a hypertensive animal model. We examined behavioral alterations (Morris water maze), lipid peroxidation (malondialdehyde (MDA)), tau and autophagy expressions, neuron death, and caspase-3 levels after 3 and 6 months of whole-body exposure to urban PM2.5 in spontaneously hypertensive (SH) rats. Results SH rats were exposed to S-, K-, Si-, and Fe-dominated PM2.5 at 8.6 ± 2.5 and 10.8 ± 3.8 μg/m3 for 3 and 6 months, respectively. We observed no significant alterations in the escape latency, distance moved, mean area crossing, mean time spent, or mean swimming velocity after PM2.5 exposure. Notably, levels of MDA had significantly increased in the olfactory bulb, hippocampus, and cortex after 6 months of PM2.5 exposure (p 2.5 caused significantly higher expressions of t-tau and p-tau in the olfactory bulb (p 2.5 exposure, but significantly decreased in the cortex with 6 months exposure to PM2.5. Neuron numbers had decreased with caspase-3 activation in the cerebellum, hippocampus, and cortex after 6 months of PM2.5 exposure. Conclusions Chronic exposure to low-level PM2.5 could accelerate the development of neurodegenerative pathologies in subjects with hypertension.

Acrylamide (AA) is a toxicant in high-temperature processed foods and an animal carcinogen. Upon absorption, AA is metabolized to glycidamide (GA) or conjugates with glutathione (AA-GSH). Important advantages of microdialysis coupled with liquid chromatography-tandem mass spectrometry (MD-LC-MS/MS) include its minimization of potential losses during sample collection, storage and preparation, as well as an improvement in temporal resolution for toxicokinetics (TKs). We aimed to simultaneously study the TKs of AA and products of its primary metabolism using an isotope-dilution (ID) MD-LC-MS/MS method. MD probes implanted into the jugular vein/right atrium of anesthetized Sprague Dawley rats were connected to the ID-LC-MS/MS for continuous monitoring of AA, GA and AA-GSH in the blood every 15 min over 8 h following intraperitoneal AA administration (0.1 mg/kg or 5 mg/kg). AA, GA, and AA-GSH TKs followed linear kinetics: GA AUC/AA AUC = 0.11 and AA-GSH AUC/AA AUC = 0.011 at 5 mg/kg. Elimination half-life (Te1/2) values were 2.44 ± 0.70, 4.93 ± 2.37 and 3.47 ± 1.47 h for AA, GA and AA-GSH, respectively. GA TKs reached a plateau at 3–6 h, suggesting that metabolic saturation of AA and Te1/2 values of the analytes were prolonged with AA at 5 mg/kg. Our results demonstrate that oxidation of AA to GA overwhelmed the conjugation of AA with GSH. Our innovative MD-ID-LC-MS/MS method facilitates the simultaneous characterization of multiple TKs associated with toxicants and their active metabolites with excellent temporal resolution to capture metabolic saturation of AA to GA.

Photoresist materials are indispensable in photolithography, a process used in semiconductor fabrication. The work process and potential hazards in semiconductor production have raised concerns as to adverse health effects. We therefore performed a health risk assessment of occupational exposure to positive photoresists in a single optoelectronic semiconductor factory in Taiwan. Positive photoresists are widely used in the optoelectronic semiconductor industry for photolithography. Occupational exposure was estimated using the Stoffenmanager® model. Bayesian modeling incorporated available personal air sampling data. We examined the composition and by-products of the photoresists according to descriptions published in the literature and patents; the main compositions assessed were propylene glycol methyl ether acetate (PGMEA), novolac resin, photoactive compound, phenol, cresol, benzene, toluene, and xylene. Reference concentrations for each compound were reassessed and updated if necessary. Calculated hazard quotients were greater than 1 for benzene, phenol, xylene, and PGMEA, indicating that they have the potential for exposures that exceed reference levels. The information from our health risk assessment suggests that benzene and phenol have a higher level of risk than is currently acknowledged. Undertaking our form of risk assessment in the workplace design phase could identify compounds of major concern, allow for the early implementation of control measures and monitoring strategies, and thereby reduce the level of exposure to health risks that workers face throughout their career.

Addressing occupational health and safety concerns early in the design stage anticipates hazards and enables health professionals to recommend control measures that can best protect workers’ health. This method is a well-established tool in public health. Importantly, its success depends on a comprehensive exposure assessment that incorporates previous exposure data and outcomes. Traditional methods for characterizing similar occupational exposure scenarios rely on expert judgment or qualitative descriptions of relevant exposure data, which often include undisclosed underlying assumptions about specific exposure conditions. Thus, improved methods for predicting exposure modeling estimates based on available data are needed. This study proposes that cluster analysis can be used to quantify the relevance of existing exposure scenarios that are similar to a new scenario. We demonstrate how this method improves exposure predictions. Exposure data and contextual information of the scenarios were collected from past exposure assessment reports. Prior distributions for the exposure distribution parameters were specified using Stoffenmanager® 8 predictions. Gower distance and k-Medoids clustering algorithm analyses grouped existing scenarios into clusters based on similarity. The information was used in a Bayesian model to specify the degree of correlation between similar scenarios and the scenarios to be assessed. Using the distance metric to characterize the degree of similarity, the performance of the Bayesian model was improved in terms of the average bias of model estimates and measured data, reducing from 0.77 (SD: 2.0) to 0.49 (SD: 1.8). Nevertheless, underestimation of exposures still occurred for some rare scenarios, which tended to be those with highly variable exposure data. In conclusion, the cluster analysis approach may enable transparent selection of similar exposure scenarios for factoring into design-phase assessments and thereby improve exposure modeling estimates.

Additional file 1: Table S1. Meteorological and gaseous data measured by the EPA Guting air quality monitoring stations during the study period. Figure S1. Spectrums of MDA analysed by LC-MS/MS. Figure S2. Effects (3- and 6-months exposure) of PM2.5 on the inflammatory infiltration of lungs in SH rats. Subpleural alveolar infiltration of mononuclear cells was observed in the lungs after 3- and 6-months exposure of HEPA and/or PM2.5. Figure S3. Alteration in body weight of SH rats between HEPA (control) and PM2.5 (exposure) groups after 6-months exposure. Figure S4. IHC images of total Tau (t-Tau) in the cerebellum, hippocampus, and cortex of SH rats after 6 months of exposure to HEPA (control) and PM2.5 (exposure). Scar bar is 50 μm. Figure S5. Chronic effects (6-months exposure) of PM2.5 on the histological changes of cerebellum, hippocampus, and cortex in SH rats. Scale bar: 200 μm.

Although the applications for zinc oxide nanoparticles (ZnONP) are continually increasing, the neurotoxicity of this material remains unclear. This study investigated the acute effects of pulmonary exposure to ZnONP on the brain in terms of behavioral changes, oxidative stress, inflammation, and tau and autophagy expressions using rat subjects. Based on the test subjects’ performance in a Morris water maze and an elevated-plus maze, respectively, no major alterations occurred in spatial cognition or learning ability, or anxiety. Following exposure to 10 mg kg–1 of ZnONP, we observed that the level of 8-hydroxy-2ʹ-deoxyguanosine (8-OHdG)/dG significantly increased in the hippocampus, whereas those of interleukin (IL)-1β and IL-6 significantly decreased in the cerebellum and the cortex. Additionally, microglia were activated in the hippocampus. Tau protein expression was strong in the cerebellum and the hippocampus, but no significant expression of Beclin-1, light chain 3 (LC3) II/I, or ubiquitin was detected. Our results suggest that acute exposure to ZnONP induces oxidative stress, microglia activation, and tau protein expression in the brain, leading to neurotoxicity.

Prenatal exposure to nonylphenol (NP) and/or bisphenol A (BPA) has been reported to be associated with adverse birth outcomes; however, the underlying mechanisms remain unclear. The primary mechanism is endocrine disruption of the binding affinity for the estrogen receptor, but oxidative stress and inflammation might also play a contributory role. We aimed to investigate urinary NP and BPA levels in relation to biomarkers of oxidative/nitrative stress and inflammation and to explore whether changes in oxidative/nitrative stress are a function of prenatal exposure to NP/BPA and inflammation in 241 mother-fetus pairs. Third-trimester urinary biomarkers of oxidative/nitrative stress were simultaneously measured, including products of oxidatively and nitratively damaged DNA (8-hydroxy-2′-deoxyguanosine (8-OHdG) and 8-nitroguanine (8-NO2Gua)) as well as products of lipid peroxidation (8-iso-prostaglandin F2α (8-isoPF2α) and 4-hydroxy-2-nonenal-mercapturic acid (HNE-MA)). The antioxidant glutathione peroxidase ...

Background Epidemiological evidence has linked fine particulate matter (PM2.5) to neurodegenerative diseases; however, the toxicological evidence remains unclear. The objective of this study was to investigate the effects of PM2.5 on neuropathophysiology in a hypertensive animal model. We examined behavioral alterations (Morris water maze), lipid peroxidation (malondialdehyde (MDA)), tau and autophagy expressions, neuron death, and caspase-3 levels after 3 and 6 months of whole-body exposure to urban PM2.5 in spontaneously hypertensive (SH) rats. Results SH rats were exposed to S-, K-, Si-, and Fe-dominated PM2.5 at 8.6 ± 2.5 and 10.8 ± 3.8 μg/m3 for 3 and 6 months, respectively. We observed no significant alterations in the escape latency, distance moved, mean area crossing, mean time spent, or mean swimming velocity after PM2.5 exposure. Notably, levels of MDA had significantly increased in the olfactory bulb, hippocampus, and cortex after 6 months of PM2.5 exposure (p

RATIONALE Maleic acid is an industrial-grade chemical that is often used in adhesives, stabilizers, and preservatives. It is unknown whether long-term consumption of maleic acid modified starch is harmful to humans. However, many studies have indicated that maleic acid causes renal tubular damage in animal models, even as the associated pathways remain unclear. Sequential window acquisition of all theoretical fragment ion spectra (SWATH) is the most innovative of the label-free quantitative technologies which have better quantification performance. Therefore, SWATH technology was used to investigate the effect of maleic acid on the rat kidney proteome in this study. METHODS Sprague-Dawley(SD) rats were treated with 0 mg/kg (control), 6 mg/kg (low-dose), 10 mg/kg (medium-dose), and 60 mg/kg (high-dose) of maleic acid. After kidney protein extraction, 28% SDS-PAGE was used, followed by in-gel digestion and desalting. Next, the samples were analyzed with ultra-performance liquid chromatography (UPLC) coupled with quadrupole time-of-flight mass spectrometry (Q-TOF MS), and data-dependent acquisition (DDA) and SWATH technology were also used. The gene ontology and pathway analysis were accomplished. Ultimately, these protein biomarkers were validated by using scheduled high-resolution multiple reaction monitoring (sMRMHR ). RESULTS Comparisons of the control group with the other three groups revealed that 95, 130, and 103 proteins were expressed at significantly different levels in the control group and in the low-dose, medium-dose, and high-dose groups, respectively. According to the gene ontology analysis, the major processes that these proteins were involved in were metabolic processes, biological regulation, cellular processes, and responses to stimuli; the major functions that these proteins were involved in were binding, hydrolase activity, catalytic activity, and oxidoreductase activity; and the major cellular components hat they were involved in were the cytoplasm, extracellular region, membrane, and mitochondria. According to the KEGG pathway analysis, these proteins were involved in 35 pathways, five of which, the carbohydrate metabolism, folate biosynthesis, renal tubular resorption, amino acid metabolism, and Ras signaling pathways, are discussed in this study. Ultimately, 19 proteins involved in 12 important pathways were validated by sMRMHR . CONCLUSIONS It was demonstrated that maleic acid caused insufficient energy production, which might lead to a decrease in the activity of the sodium-potassium ATP pump and hydrogen ion ATP pump, which could in turn have caused renal tubular resorption and hydrogen ion regulation to be blocked, thus leading to the accumulation of hydrogen ions in the renal tubules, which would then result in renal tubular acidification followed finally by Fanconi syndrome.

Pesticides are extensively used to improve crop yield and quality in many African countries where agricultural production is a critical economic activity, including Eswatini (formerly Swaziland) in southern Africa. However, the potential health effects of indoor exposure to agricultural pesticides are deeply concerning, and children are particularly vulnerable. This case study assessed such exposure and its risks by sampling and analyzing the air inside the homes of 15 pesticide applicators and 12 non-applicators in Swazi communities surrounded by nearby (~200 m) sugarcane fields on pesticide spraying days. Applying liquid chromatography with tandem mass spectrometry (LC-MS/MS) to the samples revealed mean ametryn, atrazine, pendimethalin and 2,4-dichlorophenoxyacetic acid (2,4-D) concentrations of 0.75, 0.32, 0.57 and 0.004 µg m–3, respectively, in the applicator households and corresponding values of 0.19, 0.03, 0.04 and 0.003 µg m–3, respectively, in the non-applicator households. Notably, the non-applicator households exhibited pesticide concentrations far exceeding previously published data. Furthermore, we measured significantly higher levels of ametryn and pendimethalin inside the homes of the applicators than those of the non-applicators. Daily inhalation exposure to agricultural pesticides resulted in cumulative hazard index values above 1.0 at the 95th percentile for children 3 years of age and younger in the applicator households. Thus, the take-home pathway, spray drift and household-to-field distance all play crucial roles in determining the pesticide exposure and health risks indoors. Owing to the dominance of the first factor, we advise applicators to shower and change into clean clothes before returning home to their families. This safety measure is essential for the many families living near agricultural land throughout Africa.

Maleic acid (MA) was purposefully adulterated in an array of starch-based foods in Taiwan, inciting a food safety incident. Due to limited data on the pharmacokinetics and bioavailability of ingested MA, we studied pharmacokinetic (PK) parameters in serum and urine of Sprague Dawley rats. Three groups of male and female rats were given three doses of MA by oral gavage; biofluid samples were collected accordingly. Data demonstrated that a non-compartment model best described MA's linear kinetic behavior upon ingestion. The mean residence life of maleic acid in serum was 17.58 h and 9.84 h for low-dosed male and female rats, whereas 8.24 h and 4.17 h for high-dosed male and female rats, respectively. Our results revealed oral bioavailability ranged from 30.8 to 41.0% for males and 32.2–39.1% for females. The data confirmed that ingested MA is absorbed and metabolized rapidly, along with low bioavailability. Future pathological studies may determine whether prolonged and low-level exposures of MA produce nephrotoxicity. These data provide additional contribution to current understanding of the kinetics of MA in a rat model and enable the development of a physiologically based model, which is essential to form the basis of evidenced-based food safety guidelines.

8-Hydroxy-2-deoxyguanosine (8-OHdG), 8-nitroguanine (8-NO2Gua), 8-iso-prostaglandin F2α (8-IsoPGF2α), and N-acetyl-S-(tetrahydro-5-hydroxy-2-pentyl-3-furanyl)-L-cysteine (HNE-MA) are well-studied and representative biomarkers for oxidative DNA damage, inflammation, and lipid peroxidation; all of which have been associated with increases in risks of various diseases and cancers. A rapid and highly sensitive isotope-dilution liquid-chromatography tandem mass spectrometry (LC-MS/MS) method was developed to simultaneously quantify the aforementioned biomarkers in urine. Upon validation, this method shows excellent feasibility, sensitivity (0.008-0.03 ng/mL) and satisfactory recoveries (88.7-95.4%); the calibration curves displayed excellent linearity with coefficients of determination (R(2)) greater than 0.998. Additionally, low variations were observed in the relative standard deviation for intra- and inter-day measurements for the four analytes. The relative matrix effects for all four analytes ranged from 2.04 to 3.27%, which signaled that interferences from endogenous levels of the analytes were deemed statistically insignificant. This study successfully developed an analytical method capable to simultaneously quantify urinary 8-OHdG, 8-NO2Gua, 8-IsoPGF2α, and HNE-MA. This analytical protocol can be applied towards conducting epidemiological studies to reveal the mechanisms related to disease development, and thus evaluate the associated risks of diseases.

Multiple pesticide residues are frequently present in tea leaves and while the majority of residues satisfy Taiwan's current health regulations, there are potential health effects from pesticide exposure that are of great concern for tea drinkers. We undertook a systematic probabilistic risk assessment of 59 pesticides in tea leaves from 1629 tea leaf samples obtained by Taiwan's Food and Drug Administration in two monitoring surveys in 2015. Bayesian statistics used a Markov Chain Monte Carlo approach to estimate posterior distributions of pesticide residues in tea leaves, lifetime average daily doses and hazard quotients (HQs) of evaluated pesticides. We classified 95th percentile values of HQs into three categories: 0 HQ 0.5, 0.5 ≤ HQ ≤ 1 and 1 HQ. The 95th percentiles of HQs for triazophos (3.39), carbofuran (2.04) and endosulfan (1.80) exceeded 1 in the adult population; the HQ for 3-OH carbofuran was 0.97 and was less than 0.5 for the remaining 55 pesticides. The health risk posed by pesticide residues for tea drinkers is negligible, if triazophos, carbofuran, endosulfan, and 3-OH carbofuran residues satisfy regulatory standards. However, five legacy pesticides, DDT, methomyl, carbofuran, dicofol and endosulfan, were identified. To reduce uncertainties, this study combined Bayesian statistics with a mode of action approach for systematic risk assessment of co-exposure to multiple pesticide residues in tea leaf samples. Measuring pesticide transfer rates will improve the quality of future risk assessments concerning residues in tea leaves. Appropriate management of pesticides in Taiwanese tea farms and monitoring of pesticide residues in imported tea is warranted to protect Taiwan's tea drinkers.

Maleic acid (MA), a chemical intermediate used in many consumer and industrial products, was intentionally adulterated in a variety of starch-based foods and instigated food safety incidents in Asia. We aim to elucidate possible mechanisms of MA toxicity after repeated exposure by (1) determining the changes of metabolic profile using 1H nuclear magnetic resonance spectroscopy and multivariate analysis, and (2) investigating the occurrence of oxidative stress using liquid chromatography tandem mass spectrometry by using Sprague–Dawley rat urine samples. Adult male rats were subjected to a 28 day subchronic study (0, 6, 20 and 60 mg kg−1) via oral gavage. Urine was collected twice a day on days 0, 7, 14, 21 and 28; organs underwent histopathological examination. Changes in body weight and relative kidney weights in medium- and high-dose groups were significantly different compared to controls. Morphological alterations were evident in the kidneys and liver. Metabolomic results demonstrated that MA exposure increases the urinary concentrations of 8-hydroxy-2′-deoxyguanosine, 8-nitroguanine and 8-iso-prostaglandin F2α; levels of acetoacetate, hippurate, alanine and acetate demonstrated time- and dose-dependent variations in the treatment groups. Findings suggest that MA consumption escalates oxidative damage, membrane lipid destruction and disrupt energy metabolism. These aforementioned changes in biomarkers and endogenous metabolites elucidate and assist in characterizing the possible mechanisms by which MA induces nephro- and hepatotoxicity.

Data concerning the effects of prenatal exposures to nonylphenol (NP) and oxidative stress on neonatal birth outcomes from human studies are limited. A total of 146 pregnant women were studied (1) to investigate the association between prenatal NP exposure and maternal oxidative/nitrative stress biomarkers of DNA damage (8-hydroxy-2'-deoxyguanosine (8-OHdG), 8-nitroguanine (8-NO2Gua)) and lipid peroxidation (8-iso-prostaglandin F2α (8-isoPF2α), 4-hydroxy-2-nonenal-mercapturic acid (HNE-MA)) and (2) to explore the associations among oxidative stress biomarkers, NP exposure, and neonatal birth outcomes, including gestational age, birth weight, length, Ponderal index, and head and chest circumferences. NP significantly increased the 8-OHdG and 8-NO2Gua levels. All infants born to mothers with urinary 8-OHdG levels above the median exhibited a significantly shorter gestational duration (Badjusted = -4.72 days; 95% CI: -8.08 to -1.36 days). No clear association was found between NP levels and birth outcomes. Prenatal 8-OHdG levels might be a novel biomarker for monitoring fetal health related to NP exposure.

Bisphenol A (BPA) is an endocrine disruptor widely used in the production of polycarbonate plastics and epoxy resins. Exposures to BPA have been associated with reproductive, developmental, and cardiovascular effects. In this study, the CalTOX model was used to assess the aggregate health risks on BPA by integrating the currently available BPA data in various environmental media in Taiwan. Local parameters such as chemical properties, local landscape data, and exposure factors were used as model inputs under the continuous source mode. A reference dose (RfD) of 50 μg/kg-day was adopted in this assessment. Monte Carlo simulation was used to simulate great variability of the environmental data. Our results show that an upper limit of 95 % confidence interval of aggregate exposures for the adults (19–64 years old) was 1.05 μg/kg-day, corresponding to a hazard index (HI) of 0.021. The chemical properties (BPA half-life in surface water), intake rates (fruit, vegetable, and fluid intake), and landscape data (average depth of surface waters and leaf wet density) are critical parameters. Finally, HI value would approach to 1 as BPA concentrations in ambient air, surface water, and sediment was greater than 20 ng/m3, 100 μg/L, and 3.3 mg/kg. The quality of the risk assessment on BPA can be further improved by reduction of uncertainty of the abovementioned critical parameters as well as considering additional BPA exposures from canned and packaged goods.

A rapid and simple on-line solid-phase extraction coupled with isotope dilution-liquid chromatography-tandem mass spectrometry (SPE-ID-LC-MS/MS) method was developed to quantitate maleic acid in serum and urine of SpragueDawley (SD) rats. The aforementioned biological samples were spiked with (13)C2-maleic acid, vigorously vortexed, added with acetonitrile to precipitate proteins, and then injected into the on-line SPE-LC-MS/MS system for quantification. Upon validation, this method demonstrated excellent feasibility and sensitivity: calibration curves for maleic acid in serum and urine display excellent linearity with the coefficient of determination (R(2)) greater than 0.999; the limits of detection and quantitation (LOD and LOQ) for maleic acid were determined at 0.2 and 0.5μg L(-1), respectively. Additionally, intra-day accuracy for maleic acid in serum and urine samples ranged from 94.0% to 100.2% and 101.3% to 104.4%, respectively. Furthermore, inter-day accuracy ranged from 93.6% to 101.0% and from 102.3% to 111.4% in serum and urine samples, respectively. Intra-day precision %RSD of maleic acid in serum and urine samples was 13.8% or less, whereas the inter-day precision was 6.1% or less. The matrix effects were not found to be statistically significant (p=0.9145 and p=0.5378, correspondingly) based on the calculations of recovery functions. The collected serum and urine samples were analyzed using SPE-ID-LC-MS/MS. Our results reveal trace levels of maleic acid in the control rats, demonstrating that this method is capable of analyzing background levels of contaminants in biofluids with excellent sensitivity and specificity at part-per-billion levels concentrations in complex matrices.

Combustion-derived nanoparticles are characterised by a high surface area (SA) per mass. SA is proposed to regulate the bioreactivity of nanoparticles; however, the dose metric for carbon black remains controversial. To determine the relationships between bioreactivity and SA, male spontaneously hypertensive rats were exposed to carbon black (CB) nanoparticles (15, 51 and 95 nm) via intratracheal instillation for 24 h. Pulmonary exposure to CB resulted in a significant increase in systemic 8-hydroxy-2′-deoxyguanosine (8-OHdG), DNA single-strand breakages in peripheral blood cells and pulmonary cell infiltration in rats. The oxidative potential and particularly the corresponding SA of CB were correlated with the level of 8-OHdG, DNA single-strand breakages and pulmonary cell infiltration in rats. We conclude that SA is an important dose metric for CB that can regulate oxidative stress and DNA damage in rats. Furthermore, this observation was more significant for smaller sized CB.

Maleic acid (MA), an intermediate reagent used in many industrial products, instigated public health concerns in Taiwan when it was used to adulterate an array of starch-based delicacies to improve texture and storage time. Established studies reported that exposure to high concentrations of MA induce renal injury; little is known whether oxidative stress is induced at a relative low dose. This study aims to investigate the effect of oral single dose exposure of MA on the status of oxidative stress and inflammation. Single dose of MA at 0, 6 and 60 mg/kg (control, low- and high-dose groups, respectively) were orally administered to adult male and female rats. Urine samples were collected and analyzed to measure 8-hydroxy-2'-deoxyguanosine (8-OHdG), 8-iso-prostaglandin F2α (8-IsoPGF2α), 8-nitroguanine (8-NO2Gua) and N-acetyl-S-(tetrahydro-5-hydroxy-2-pentyl-3-furanyl)-L-cysteine (HNE-MA) using LC-MS/MS. Results revealed that oral consumption of MA induced oxidative DNA damage and lipid peroxidation, as demonstrated by the statistically significant increases in urinary levels of 8-NO2Gua, 8-OHdG, and 8-isoPGF2α, in high-dosed male rats within 12 h of oral gavage (p < 0.05). Additionally, increases in concentration of these biomarkers persist for days after consumption; male rats appear to be more sensitive to oxidative burden compared to their counterparts. The aforementioned findings could help elucidate the mechanisms through which nephrotoxicity occur.

Acrylamide (AA), a probable human carcinogen, is a widely-used industrial chemical but is also present in tobacco smoke and carbohydrate-rich foods processed at high temperatures. AA is metabolized to glycidamide (GA) to cause the formation of DNA adducts. N7-(2-carbamoyl-2-hydroxyethyl) guanine (N7-GAG), the most abundant DNA adduct induced by GA, was recently detected in urine of smokers and non-smokers. In this study, we assessed the variability of AA exposure and biomarkers of AA exposure in urine samples repeatedly collected from AA-exposed workers and explored the half-life of N7-GAG. A total of 8 AA-exposed workers and 36 non-exposed workers were recruited. Pre-shift and post-shift urine samples were collected from the exposed group in parallel with personal sampling for eight consecutive days and from the control group on day 1 of the study. Urinary N7-GAG and the mercapturic acids of AA and GA, namely N-acetyl-S-(2-carbamoylethyl)-L-cysteine (AAMA) and N-(R,S)-acetyl-S-(1-carbamoyl-2-hydroxyethyl)-L-cysteine (GAMA) were analyzed using on-line solid phase extraction-liquid chromatography-electrospray ionization/tandem mass spectrometry methods. We found that N7-GAG levels in urine were significantly higher in exposed workers than in controls and that N7-GAG level correlated positively with AAMA and GAMA levels. Results from this study showed that AAMA and GAMA possibly remain the more preferred biomarkers of AA exposure and that N7-GAG levels could be elevated by occupational exposures to AA and serve as a biomarker of AA-induced genotoxicity for epidemiological studies.

A two-dimensional (2D) hydrophilic interaction liquid chromatography (HILIC) and reverse-phase (RP) liquid chromatography (LC) system coupled with triple-quadrupole mass spectrometry (MS) was developed to comprehensively profile ceramides and phosphatidylcholine in extracted biological samples. Briefly, the 2D HILIC-RPLC system used a silica HILIC column operated in the first dimension to distinguish the lipid classes and a BEH C18 column operated in the second dimension to separate the lipid species of the same class. The regression linearity of each lipid was satisfactory in both systems; however, the absolute matrix effect factor was reduced in 2D LC-MS/MS system. Limits of detection of 2D LC-MS/MS system were 2- to 3-fold lower compared with one-dimensional RPLC-MS/MS. The recovery from the sample ranged from 84.5 to 110%. To summarize, the developed method was proven to be accurate and producible, as relative standard deviations remained

We evaluated the reliability of 8-hydroxy-2'-deoxyguanosine (8-OHdG), and determined its ability to predict functional outcomes in stroke survivors. The rehabilitation effect on 8-OHdG and functional outcomes were also assessed. Sixty-one stroke patients received a 4-week rehabilitation. Urinary 8-OHdG levels were determined by liquid chromatography–tandem mass spectrometry. The test-retest reliability of 8-OHdG was good (interclass correlation coefficient = 0.76). Upper-limb motor function and muscle power determined by the Fugl-Meyer Assessment (FMA) and Medical Research Council (MRC) scales before rehabilitation showed significant negative correlation with 8-OHdG (r = −0.38, r = −0.30; p < 0.05). After rehabilitation, we found a fair and significant correlation between 8-OHdG and FMA (r = −0.34) and 8-OHdG and pain (r = 0.26, p < 0.05). Baseline 8-OHdG was significantly correlated with post-treatment FMA, MRC, and pain scores (r = −0.34, −0.31, and 0.25; p < 0.05), indicating its ability to predict functional outcomes. 8-OHdG levels were significantly decreased, and functional outcomes were improved after rehabilitation. The exploratory study findings conclude that 8-OHdG is a reliable and promising biomarker of oxidative stress and could be a valid predictor of functional outcomes in patients. Monitoring of behavioral indicators along with biomarkers may have crucial benefits in translational stroke research.

Perfluoroalkyl substances (PFASs) can cross the placenta, enter fetal circulation, and were found to correlate with adverse fetal growth. However, determinants of cord blood PFASs are not fully characterized. The study aimed to explore the association between PFASs and neonatal-maternal factors within a Taiwanese birth cohort. We selected subjects from Taiwan Birth Panel Study, which enrolled 486 infant-mother pairs in 2004-2005. We collected cord blood and analyzed perfluorooctanoic acid (PFOA), perfluorooctanyl sulfonate (PFOS), perfluorononanoic acid (PFNA) and perfluoroundecanoic acid (PFUA) using a simple protein precipitation and an ultra-high performance liquid chromatography/tandem mass spectrometry. We retrieved information pertaining to maternal socio-demographics, lifestyle- and dietary-related factors through structured questionnaires during the postpartum hospital stay. A total of 439 subjects, with 90% response rate, have completed serum analysis and questionnaire survey. The median concentrations for PFOA, PFOS, PFNA, and PFUA in cord blood were 1.86, 5.67, 3.00, and 13.5ngmL(-1), respectively. After adjusting for potential confounders, multiple linear regression models revealed that log10-PFOA was positively associated with maternal age (β=0.011) and negatively associated with multiparity (β=-0.044). Log10-PFOS was negatively correlated with birth weight (β=-0.011) and higher maternal education (senior high school: β=-0.067; university: β=-0.088). Log10-PFUA tended to negatively associate with gender, male infants (β=-0.075), and using cosmetics during pregnancy (β=-0.065). Interestingly, presence of cockroaches in the home was positively associated with log10-PFOA (β=0.041) and 1og10-PFNA (β=0.123). In conclusion, this study demonstrated several factors to correlate with cord blood PFASs and further investigation are still needed for confirmation of exposure routes.

Introduction Environmental tobacco smoke (ETS) is a hazardous component of indoor air, and may increase the risk of respiratory diseases, atherosclerosis and otitis media in children. In this study, we explored the relationship between time inside the house, ETS exposure and urinary cotinine level, and also determined the association of time inside the house on asthma phenotypes when children exposed to ETS. Methods A total of 222 asthmatic children and 205 non-asthmatic controls were recruited in the Genetic and Biomarker study for Childhood Asthma (GBCA). Structured questionnaires and time–location pattern questionnaires were administered by face-to-face interview. Urinary cotinine was measured by liquid chromatography tandem mass spectrometry (LC/MS/MS). The level of household ETS exposure was assessed using the cotinine/creatinine ratio (CCR). Results In general, urinary cotinine and CCR were higher in subjects exposed to household ETS than those who never had ETS at home. A significant positive relationship was found between average time inside the house and urinary CCR in asthmatic children with current ETS at home (β=0.278, p=0.02). After adjustment for age and gender, average time inside the house was positively related to severe wheeze in asthmatic children with household ETS within 1 month (OR: 1.26, 95%: 1.02–1.64). Conclusions Our study suggests that the major source of ETS exposure for children is due to longer period of exposures among children living with adult smokers at home. Home-smoking restrictions that effectively prevent children from being exposed to ETS would be worthwhile.

WOS: 000319871200001, PubMed ID: 23198723, Aims: Urinary 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) is a widely used biomarker of oxidative stress. However, variability between chromatographic and ELISA methods hampers interpretation of data, and this variability may increase should urine composition differ between individuals, leading to assay interference. Furthermore, optimal urine sampling conditions are not well defined. We performed inter-laboratory comparisons of 8-oxodG measurement between mass spectrometric-, electrochemical- and ELISA-based methods, using common within-technique calibrants to analyze 8-oxodG-spiked phosphate-buffered saline and urine samples. We also investigated human subject- and sample collection-related variables, as potential sources of variability. Results: Chromatographic assays showed high agreement across urines from different subjects, whereas ELISAs showed far more inter-laboratory variation and generally overestimated levels, compared to the chromatographic assays. Excretion rates in timed 'spot' samples showed strong correlations with 24 h excretion (the 'gold' standard) of urinary 8-oxodG (r(p) 0.67-0.90), although the associations were weaker for 8-oxodG adjusted for creatinine or specific gravity (SG). The within-individual excretion of 8-oxodG varied only moderately between days (CV 17% for 24 h excretion and 20% for first void, creatinine-corrected samples). Innovation: This is the first comprehensive study of both human and methodological factors influencing 8-oxodG measurement, providing key information for future studies with this important biomarker. Conclusion: ELISA variability is greater than chromatographic assay variability, and cannot determine absolute levels of 8-oxodG. Use of standardized calibrants greatly improves intra-technique agreement and, for the chromatographic assays, importantly allows integration of results for pooled analyses. If 24 h samples are not feasible, creatinine- or SG-adjusted first morning samples are recommended., ECNIS (Environmental Cancer Risk, Nutrition and Individual Susceptibility), a network of excellence operating within the European Union 6th Framework Program, Priority 5:"Food Quality and Safety" [FOOD-CT-2005-513943]; ECNIS2, a coordination and support action within the European Union FP7 Cooperation Theme 2 Food, Agriculture, Fisheries and Biotechnologies; CISBO; Ingeborg; Leo Dannin Foundation; National Science Council, TaiwanNational Science Council of Taiwan [NSC 97-2314-B-040-015-MY3, NSC 100-2628-B-040-001-MY4]; US NIHUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USA [P30ES009089]; Instituto Carlos III division of the Government for Clinical Research [PI-10/00802, RD06/0045/0006]; Generalitat ValencianaGeneralitat Valenciana [ACOM/2012/238]; Swedish Council for Working Life and Social ResearchSwedish Research CouncilSwedish Research Council for Health Working Life & Welfare (Forte); TUBITAK (Technical and Scientific Research Council of Turkey)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [108Y049]; Grant Agency of the Czech RepublicGrant Agency of the Czech Republic [P503/11/0084]; Sahlgrenska University Hospital, Gothenburg; UK Medical Research Council via a People Exchange Programme Research Leader Fellowship award [G1001808/98136], Some of the authors of this work were partners in, and this work was partly supported by, ECNIS (Environmental Cancer Risk, Nutrition and Individual Susceptibility), a network of excellence operating within the European Union 6th Framework Program, Priority 5:"Food Quality and Safety" (Contract No. FOOD-CT-2005-513943), and also ECNIS2, a coordination and support action within the European Union FP7 Cooperation Theme 2 Food, Agriculture, Fisheries and Biotechnologies.; P Moller and S Loft are supported by CISBO and the Ingeborg and Leo Dannin Foundation.; M-R Chao and C-W Hu acknowledge financial support from the National Science Council, Taiwan (Grants NSC 97-2314-B-040-015-MY3 and NSC 100-2628-B-040-001-MY4).; R Santella acknowledges the contribution of Qiao Wang, and support from US NIH P30ES009089.; G Saez and C Cerda acknowledge financial support from the Instituto Carlos III division of the Government for Clinical Research (Grants PI-10/00802 and RD06/0045/0006) and Grant ACOM/2012/238 from Generalitat Valenciana.; K Broberg, C Lindh, and M Hossain acknowledge financial support from the Swedish Council for Working Life and Social Research; H Orhan and N Senduran acknowledge financial support from TUBITAK (Technical and Scientific Research Council of Turkey), grant number 108Y049.; P Rossner, Jr. and RJ Sram acknowledge support from the Grant Agency of the Czech Republic (P503/11/0084).; L Barregard acknowledges financial support from the Sahlgrenska University Hospital, Gothenburg.; MS Cooke acknowledges support from the UK Medical Research Council via a People Exchange Programme Research Leader Fellowship award (G1001808/98136).

To date, the variant Creutzfeldt-Jakob disease (vCJD) risk assessments that have been performed have primarily focused on predicting future vCJD cases in the United Kingdom, which underwent a bovine spongiform encephalopathy (BSE) epidemic between 1980 and 1996. Surveillance of potential BSE cases was also used to assess vCJD risk, especially in other BSE-prevalent EU countries. However, little is known about the vCJD risk for uninfected individuals who accidentally consume BSE-contaminated meat products in or imported from a country with prevalent BSE. In this article, taking into account the biological mechanism of abnormal prion PrP(res) aggregation in the brain, the probability of exposure, and the expected amount of ingested infectivity, we establish a stochastic mean exponential growth model of lifetime exposure through dietary intake. Given the findings that BSE agents behave similarly in humans and macaques, we obtained parameter estimates from experimental macaque data. We then estimated the accumulation of abnormal prions to assess lifetime risk of developing clinical signs of vCJD. Based on the observed number of vCJD cases and the estimated number of exposed individuals during the BSE epidemic period from 1980 to 1996 in the United Kingdom, an exposure threshold hypothesis is proposed. Given the age-specific risk of infection, the hypothesis explains the observations very well from an extreme-value distribution fitting of the estimated BSE infectivity exposure. The current BSE statistics in the United Kingdom are provided as an example.

Although, humans are exposed to perflourinated compounds (PFCs) from various media, water consumption could be an important source for the residents living near to contaminated areas. Since comprehensive multimedia exposure model has not been developed for PFCs, assessment of the potential risk due to exposure to PFCs through direct water consumption could be a conservative estimate. The human health risks derived from the exposure to PFCs through water consumption were assessed for different age groups of general population in Taiwan using probabilistic approach. Based on available data on concentrations of PFCs in river water, exposure to PFOS, PFOA and PFDA via water consumption for different age groups were calculated using deterministic and probabilistic risk assessment methods. The oral non-cancer risks from PFOS, PFOA and their combination, expressed as a Hazard Index (HI), was determined by comparing oral exposure dose (through water intake) with the oral Reference Dose (RfD). The average exposure to PFOS, PFOA and PFDA via water consumption for adults ranged from 0.16 to 220.15, 0.43 to 12.5 and 0.43 to 2.36 ng kg-bw−1 d−1 and for children 0.13–354.3, 0.35–20.17 and 0.35–3.79 ng kg-bw−1 d−1, respectively. Probabilistic values of total HIs for all age groups reside near to Keya River exceed the RfD 2.4–4.8 times, corresponding mainly to PFOS with a percentage of 97%. In conclusions, children aged 1–3 years old and the residents reside near to Keya River are at the highest risk of exposure to PFCs via water consumption.

Introduction: No study have comprehensively evaluated the complex pollutants emitted from coal-fired power plants and oil refineries in petrochemical industries and the health impact on nearby comm...

This study assesses the association of acrylamide (AA) and glycidamide (GA) hemoglobin adducts (AAVal and GAVal) and their ratios with genetic polymorphisms of the metabolic enzymes cytochrome P450 2E1 (CYP2E1), exon 3 and 4 of microsomal epoxide hydrolase (mEH3 and mEH4), glutathione transferase theta (GSTT1), and mu (GSTM1) or/and the combinations of these polymorphisms, involved in the activation and detoxification of AA in humans. Fifty-one AA-exposed workers and 34 controls were recruited and provided a post-shift blood sample. AAVal and GAVal were determined simultaneously using isotope-dilution liquid chromatography-electronspray ionization/tandem mass spectrometry (LC-ESI-MS/MS). Genetic polymorphisms of CYP2E1, mEH3 and 4, GSTT1, and GSTM1 were also analyzed. Our results reveal that the GAVal/AAVal ratio, potentially reflecting the proportion of AA metabolized to GA, ranged from 0.13 to 0.45 with a mean at 0.27. Multivariate regression analysis demonstrates that the joint effect of CYP2E1, GSTM1, and mEH4 genotypes was significantly associated with AAVal and GAVal levels after adjustment for AA exposures. These results suggest that mEH4 and the combined genotypes of CYP2E1, GSTM1 and mEH4 may be associated with the formation of AAVal and GAVal. Further studies may be needed to shed light on the roles that phase I and II enzymes play in AA metabolism.

Safrole, a naturally occurring product derived from spices and herbs, has been shown to be associated with the development of hepatocellular carcinoma in rodents. Safrole 2',3'-oxide (SFO), an electrophilic metabolite of safrole, was shown to react with DNA bases to form detectable DNA adducts in vitro, but not detected in vivo. Therefore, the objective of this study was to investigate the formation of N7-(3-benzo[1,3]dioxol-5-yl-2-hydroxypropyl)guanine (N7γ-SFO-Gua) resulting from the reaction of SFO with the most nucleophilic site of guanine in vitro and in vivo with a newly developed isotope-dilution high performance liquid chromatography electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS) method. N7γ-SFO-Gua and [(15)N(5)]-N7-(3-benzo[1,3]dioxol-5-yl-2-hydroxypropyl)guanine ([(15)N(5)]-N7γ-SFO-Gua) were first synthesized, purified, and characterized. The HPLC-ESI-MS/MS method was developed to measure N7γ-SFO-Gua in calf thymus DNA treated with 60 μmol of SFO for 72 h and in urine samples of mice treated with a single dose of SFO (30 mg/kg body weight, intraperitoneally). In calf thymus DNA, the level of N7γ-SFO-Gua was 2670 adducts per 10(6)nucleotides. In urine of SFO-treated mice, the levels of N7γ-SFO-Gua were 1.02±0.14 ng/mg creatinine (n=4) on day 1, 0.73±0.68 ng/mg creatinine (n=4) on day 2, and below the limit of quantitation on day 3. These results suggest that SFO can cause in vivo formation of N7γ-SFO-Gua, which may then be rapidly depurinated from the DNA backbone and excreted through urine.

Fingerprint analysis comparing the polychlorinated dibenzo-p-dioxin and dibenzofuran (PCDD/F) congener profile patterns of collected samples with those of potential dioxin emission source(s) is an important tool for identifying environmental dioxin pollution. The constraint that the proportions of the 17 PCDD/F congeners comprising a fingerprint sum up to one motivates a multivariate gamma distribution, which leads to a Dirichlet distribution. Because of the complexity in restricted likelihood ratio tests and typical sample size limitations resulting from laboratory analysis costs, permutation test procedures are employed for hypothesis testing of the homogeneity of congener profiles. Pearson-type chi-squared tests based on the Dirichlet distribution (DM) assumption, the generalized form of DM using the arithmetic mean and geometric mean of the proportions, and the robust aligned rank test, are proposed and compared through simulations. PCDD/F samples collected from the stack of a local municipal solid waste incinerator and from ambient air near the municipal solid waste incinerator in Taiwan were illustrated as an example. The simulation results showed that the aligned rank test, followed by the DM-based test, was generally robust to distributional assumptions and had high statistical power. The arithmetic-mean-based and geometric-mean-based tests outperformed one another in different conditions, dependent on the underlying distribution. Copyright © 2012 John Wiley & Sons, Ltd.

Safrole (1) is a natural product found in herbs and spices. Upon uptake, it can be metabolized to safrole 2′,3′-oxide [(±)-SFO, 2], which can react with DNA bases to form DNA adducts. The reactions of 2 with 2′-deoxyadenosine (3) and adenine (8) under physiological conditions (pH 7.4, 37 °C) were carried out to characterize its possible adducts with adenine. Four adducts were isolated by reverse-phase liquid chromatography and their structures were characterized by UV/Vis, 1H and 13C NMR spectroscopy and MS. The reaction of 2 with 3 produced two regioisomers, N1γ-SFO-dAdo (4) and N6γ-SFO-dAdo (5), in 4.2–4.5 % yield, and the reaction of 2 with 8 generated N3γ-SFO-Ade (9) and N9γ-SFO-Ade (10) in 1.0–2.4 % yield. Using HPLC–ESI-MS/MS, we traced the amounts of the four adducts formed when calf thymus DNA (10 mg) was treated with 2 (60 μmol) and the levels of 4, 5, and 9 were determined to be 2000, 170, and 660 adducts per 106 nucleotides, respectively. Adduct 10 was not detected under these conditions. These results suggested that stable DNA adducts of 2 were formed in vitro, and further studies on the formation of these DNA adducts in vivo may help to elucidate their role in safrole carcinogenicity.

The total surface area is known to be an effective exposure metric for predicting the lung toxicity of low solubility nanoparticles (NPs). However, if NPs are dissolved quickly enough in the lungs, the mass may be correlated with the toxicity. Recent studies have found that the toxicity of zinc oxide (ZnO) NPs was caused by the release of zinc ions. Thus, we hypothesized that mass could be used as an exposure metric for the toxicity of ZnO NPs. Healthy Sprague-Dawley rats were exposed to a low, moderate, or high dose of 35 and 250 nm ZnO particles or filtered air. Bronchoalveolar lavage fluid was collected to determine lung inflammation, injury and oxidative stress. The lung inflammation induced by ZnO particles according to different concentration metrics, including number, mass and surface area, was compared. The mass concentration was significantly correlated with the percentage of neutrophils (R(2) = 0.84), number of neutrophils (R(2) = 0.84) and total cells (R(2) = 0.73). Similarly, surface area concentration was significantly correlated with the percentage of neutrophils (R(2) = 0.94), number of neutrophils (R(2) = 0.81) and total cells (R(2) = 0.76). There was no correlation between the number and lung inflammation. We found that both mass and surface area were effective as metrics for the toxicity of ZnO NPs, although only surface area was previously indicated to be an effective metric. Our results are also consistent with recent study results that ZnO NPs and released zinc ions may play a role mediating the toxicity of NPs.

Background The effects of emissions from coal-fired power plants on children's exposure to polycyclic aromatic hydrocarbons (PAHs) are not well understood. Objectives This study aimed to evaluate the sources and the urinary levels of 1-hydroxypyrene, a biomarker of exposure to PAHs, among children living in proximity to a coal-fired power plant. Methods Study areas consisted of two high exposure and two low exposure communities, at different distances and directions from a large coal-fired power plant in central Taiwan. Study subjects included 369 children aged 1–13 years and randomly selected from each community. Each child's urinary 1-hydroxypyrene concentration was measured by a high-performance liquid chromatography-fluorescence detector method. Samples of ambient air were analyzed for PAHs using a gas chromatography–mass spectrometry method. Information on important factors was collected by an interview using a structured questionnaire. Multiple regression analysis was used to assess factors significantly associated with urinary 1-hydroxypyrene levels. Results Levels of PAHs in ambient air in the high exposure communities were higher than those in the low exposure communities. Children living in high exposure communities had higher mean urinary 1-hydroxypyrene concentrations than those in low exposure communities (0.186 and 0.194 vs. 0.113 and 0.122 μmol/mol-creatinine, respectively). The difference in urinary 1-hydroxypyrene levels between the high exposure communities and one low exposure community remained significant after adjusting for age, gender, environmental tobacco smoke, dietary exposure, and traffic. Conclusions Children living in communities downwind of and in proximity to the coal-fired power plant had significantly increased urinary 1-hydroxypyrene levels.

Aristolochic acids (AAs), nephrotoxicants and known human carcinogens, are a mixture of structurally related derivatives of nitrophenanthrene carboxylic acids with the major components being aristolochic acid I and aristolochic acid II. People may ingest small amounts of AAs from its natural presence in medicinal plants and herbs of the family Aristolochiaceae, including the genera Aristolochia and Asarum, which have been used worldwide in folk medicine for centuries. In order to assess AA intake, an on-line solid-phase extraction coupled with liquid chromatography–tandem mass spectrometry (on-line SPE-LC/MS/MS) method was developed to analyze their most abundant corresponding metabolites, aristolactams (ALs), in urine to serve as biomarkers. The limits of quantitation were 0.006 ng for aristolactam I (AL-I), and 0.024 ng for aristolactam II (AL-II) on column. Recovery varied from 98.0% to 99.5%, and matrix effects were within 75.3–75.4%. This method was applied to analyze ALs in the urine samples collected on days 1, 2, 4, and 7 from mice treated with 30 mg/kg or 50 mg/kg AAs. Their half lives were estimated to be 3.55 h and 4.00 for AL-I, and 4.04 and 4.83 h for AL-II, depending on AAs doses. These results demonstrated that the first simple on-line SPE-LC/MS/MS method was successfully developed to analyze urinary ALs with excellent sensitivity and specificity to serve as biomarkers to assess current AA intake from AAs-containing Chinese herbs.

Chlorpyrifos and cypermethrin are the most used insecticides in Taiwan. Exposure to both pesticides has been associated with reproductive and developmental health effects in humans and animals. This study describes an online solid-phase extraction coupled with liquid chromatography-heated electrospray ionization tandem mass spectrometry (online SPE-LC/HESI/MS/MS) method to analyze chlorpyrifos and cypermethrin in cord blood of pregnant women. Calibration curves showed good linearity (r² > 0.998) for both pesticides within the range of 0.1-100 ppb. Limits of detection (LODs) were 0.01 and 0.05 ppb and recoveries in cord blood were 97.2 ± 4.8% and 93.5 ± 9.5% for chlorpyrifos and cypermethrin respectively. After analysis of 396 samples, the mean concentrations of chlorpyrifos and cypermethrin were 0.38 and 1.08 ppb respectively. These results demonstrate that LC/HESI/MS/MS is effective for the simultaneous analysis of chlorpyrifos and cypermethrin in cord blood with excellent sensitivity and specificity and may also be effective for high throughput assay in future epidemiology studies.

The use of a physiologically based toxicokinetic (PBTK) model to reconstruct chemical exposure using human biomonitoring data, urinary metabolites in particular, has not been fully explored. In this paper, the trichloroethylene (TCE) exposure dataset by Fisher et al. (Toxicol Appl Pharm 152:339–359, 1998) was reanalyzed to investigate this new approach. By treating exterior chemical exposure as an unknown model parameter, a PBTK model was used to estimate exposure and model parameters by measuring the cumulative amount of trichloroethanol glucuronide (TCOG), a metabolite of TCE, in voided urine and a single blood sample of the study subjects by Markov chain Monte Carlo (MCMC) simulations. An estimated exterior exposure of 0.532 mg/l successfully reconstructed the true inhalation concentration of 0.538 mg/l with a 95% CI (0.441–0.645) mg/l. Based on the simulation results, a feasible urine sample collection period would be 12–16 h after TCE exposure, with blood sampling at the end of the exposure period. Given the known metabolic pathway and exposure duration, the proposed computational procedure provides a simple and reliable method for environmental (occupational) exposure and PBTK model parameter estimation, which is more feasible than repeated blood sampling.

This study elucidates the association of acrylamide metabolites, N-acetyl-S-(2-carbamoylethyl)-cysteine (AAMA), N-acetyl-S-(1-carbamoyl-2-hydroxyethyl)-cysteine (GAMA2), and N-acetyl-S-(2-carbamoyl-2-hydroxyethyl)-cysteine (GAMA3) in urine with genetic polymorphisms of the metabolic enzymes cytochrome P450 2E1 ( CYP2E1 ), microsomal epoxide hydrolase ( mEH ) in exon 3 and exon 4, glutathione transferase theta ( GSTT1 ) and mu ( GSTM1 ), involved in the activation and detoxification of acrylamide (AA) in humans. Eighty-five workers were recruited, including 51 AA-exposed workers and 34 administrative staffs serve as controls. Personal air sampling was performed for the exposed workers. Each subject provided pre- and post-shift urine samples and blood samples. Urinary AAMA, GAMA2 and GAMA3 levels were simultaneously quantified using liquid chromatography-electronspray ionization/tandem mass spectrometry (LC-ESI–MS/MS). CYP2E1 , mEH (in exon 3 and exon 4), GSTT1 , and GSTM1 were analyzed using polymerase chain reaction (PCR). Our results reveal that AA personal exposures ranged from 4.37 × 10 −3 to 113.61 μg/m 3 with a mean at 15.36 μg/m 3 . The AAMA, GAMA2, and GAMA3 levels in the exposed group significantly exceeded those in controls. The GAMAs (the sum of GAMA2 and GAMA3)/AAMA ratios, potentially reflecting the proportion of AA metabolized to glycidamide (GA), varied from 0.003 to 0.456, and indicate high inter-individual variability in the metabolism of AA to GA in this study population. Multivariate regression analysis demonstrates that GSTM1 genotypes significantly modify the excretion of urinary AAMA and the GAMAs/AAMA ratio, exon 4 of mEH was significantly associated with the urinary GAMAs levels after adjustment for AA exposures. These results suggest that mEH and/or GSTM1 may be associated with the formation of urinary AAMA and GAMAs. Further study may be needed to shed light on the role of both enzymes in AA metabolism.

Mass spectrometry plays an increasingly important role in the search for and quantification of novel chemically specific biomarkers. The revolutionary advances in mass spectrometry instrumentation and technology empower scientists to specifically analyze DNA and protein adducts, considered as molecular dosimeters, derived from reactions of a carcinogen or its active metabolites with DNA or protein. Analysis of the adducted DNA bases and proteins can elucidate the chemically reactive species of carcinogens in humans and can serve as risk-associated biomarkers for early prediction of cancer risk. In this article, we review and compare the specificity, sensitivity, resolution, and ease-of-use of mass spectrometry methods developed to analyze ethylene oxide (EO)-induced DNA and protein adducts, particularly N7-(2-hydroxyethyl)guanine (N7-HEG) and N-(2-hydroxyethyl)valine (HEV), in human samples and in animal tissues. GC/ECNCI-MS analysis after HPLC cleanup is the most sensitive method for quantification of N7-HEG, but limited by the tedious sample preparation procedures. Excellent sensitivity and specificity in analysis of N7-HEG can be achieved by LC/MS/MS analysis if the mobile phase, the inlet (split or splitless), and the collision energy are properly optimized. GC/ECNCI-HRMS and GC/ECNCI-MS/MS analysis of HEV achieves the best performance as compared with GC/ECNCI-MS and GC/EI-MS. In conclusion, future improvements in high-throughput capabilities, detection sensitivity, and resolution of mass spectrometry will attract more scientists to identify and/or quantify novel molecular dosimeters or profiles of these biomarkers in toxicological and/or epidemiological studies.