Diethyl Thiophosphate Research Articles

A green and sensitive electromembrane extraction (EME) method for the determination of dialkylphosphates in maternal urine and amniotic fluid

Organophosphates compounds are extensively used pesticides, whose metabolites, dialkyl phosphates, are considered as bioindicators of human exposure. Recently, research has indicated that they can affect thyroid endocrine function. Pregnant women are a particularly vulnerable cohort to these effects due to concerns about prenatal exposure to environmental pollutants. For the first time an electromembrane microextraction method is proposed for the simultaneous determination of seven dialkylphosphate metabolites in maternal urine and amniotic fluid, which is compatible with mass spectrometry detection and achieves highly sensitive levels of quantitation. Dialkylphosphates were extracted from a 10 mL donor solution at pH 7 into a 50 µL acceptor solution at pH 12 using 1-octanol as the supported liquid membrane and applying 60 V for 10 min at 600 rpm. The obtained extracts were analysed by ion pair liquid chromatography coupled to a triple quadrupole mass spectrometer, using an Acquity UPLC® BEH C18 column (100 mm × 2.1 mm i.d., 1.7 µm particle size) at 30 °C. Under optimal extraction conditions, the preconcentration factors were achieved were up to 178-fold, allowing for high sensitivity levels, in the order of ng mL−1. The limits of quantitation were within 0.075 ng mL−1 (diethyl thiophosphate) and 1.67 ng mL−1 (dimethyl phosphate) in maternal urine samples and within 0.015 ng mL−1 (diethyl thiophosphate) and 0.5 ng mL−1 (dimethyl dithiophosphate) in amniotic fluid samples. The comparative study revealed a significant improvement in terms of analysis time, sensitivity, greenness, and practicality. Finally, the applicability of the method for the determination of selected dialkylphosphates was demonstrated in paired maternal urine and amniotic fluid samples. These results suggest potential placental transfer to the offspring.

Enhanced efficiency and stability in the degradation of triazophosphorus pesticides by Al6Si2O13/WO2.72 nanocomposites through synergistic action of S-scheme heterojunction and oxygen vacancies

The environmental contamination caused by organophosphorus pesticides (for example, triazophos) is an escalating concern. To mitigate this issue, this study introduces a novel Al6Si2O13/WO2.72 (ASO/WO) nanocomposite photocatalyst, which markedly enhances the photocatalytic degradation of triazophos. The optimized nanocomposite material with a 60.0 % ASO loading (60-ASO/WO) achieves a degradation rate of 86.3 % for triazophos within 140.0 min, marginally exceeding 60-ASO/WO3 (72.6 %) and significantly outperforming individual ASO (65.0 %), WO (59.5 %), and WO3 (56.2 %). This catalyst retains 88.9 % of its activity after five cycles, showcasing exceptional efficiency and stability. Additionally, its electrochemical surface area (ECSA, 310.0 cm2), total organic carbon (TOC, removal rate of 73.7 %), photocurrent, and electrochemical impedance are all optimal. X-ray photoelectron spectroscopy (XPS), electron paramagnetic resonance (EPR), and theoretical calculations elucidate the critical role of oxygen vacancies and the S-scheme heterojunction in augmenting charge separation and photocatalytic performance, corroborating the synergistic effect of oxygen defects and the S-scheme. While individual factors can enhance photocatalytic activity, their combination results in a more pronounced effect. Liquid chromatography-mass spectrometry (LCMS) identifies the principal degradation intermediates, including 1-phenyl-3-hydroxy-1, 2, 4-triazole, diethyl thiophosphate, and 3, 5, 6-trichloro-2-pyridinol, underscoring the material’s potential in environmental remediation.

Determinants of urinary dialkyl phosphate metabolites in midlife women: the Study of Women's Health Across the Nation Multi-Pollutant Study (SWAN-MPS).

Biomonitoring data and determinants of urinary dialkylphosphate (DAP) metabolites, markers of organophosphate pesticides, in racially diverse, non-occupationally exposed populations are scarce. This study evaluated urinary concentrations and potential determinants of DAP metabolites of organophosphate pesticides in a multi-site, multi-racial/ethnic cohort of women aged 45-56 years, the Study of Women's Health Across the Nation Multi-Pollutant Study (SWAN-MPS). We analyzed 963 urine samples collected in 1999-2000, the baseline of SWAN-MPS for longitudinal studies, and quantified DAP metabolites, including dimethyl alkylphosphates (DMAPs): dimethylphosphate (DMP), dimethylthiophosphate (DMTP), dimethyldithiophosphate (DMDTP); and diethyl alkylphosphates (DEAPs): diethylphosphate (DEP), diethylthiophosphate (DETP), diethyldithiophosphate (DEDTP), using gas chromatography and triple quadrupole mass spectroscopy. Adjusted least squared geometric means (LSGMs) and 95% confidence intervals (CIs) were computed to compare DAP concentrations by socio-demographic, behavioral and dietary factors. The geometric means (geometric standard deviations) of total DAPs, DMAPs, and DEAPs were 141 (2.63) nmol/L, 102 (2.99) nmol/L, and 26.8 (2.46) nmol/L, respectively. Body mass index (BMI) was inversely associated with DMAPs and DEAPs: LSGM (95% CI) = 68.8 (55.7-84.9) and 21.0 (17.7-25.0) nmol/L for women with obesity vs. 102 (84.7-123) and 30.1 (25.7-35.1) nmol/L for women with normal/underweight, respectively. Fruit consumption was positively (74.9 (62.1-90.2) for less than 5-6 servings/week vs. 105 (84.8-130) nmol/L for 1 serving/day and more) whereas meat consumption was inversely associated with DMAPs (110 (95.0-128) for seldom vs. 82.3 (59.5-114) nmol/L for often consumption). Fresh apple consumption appears to be attributed to the DMAP differences. Alcohol consumption was positively associated with DEAPs (27.5 (23.1-32.7) for 2 drinks/week and more vs. 23.0 (20.0-26.6) nmol/L for less than 1 drink/month). Black women had higher concentrations of DEAPs compared with White women (27.3 (21.2-35.2) vs. 23.2 (20.2-26.7) nmol/L). Organophosphate pesticides (OPs) are synthetic chemicals and currently the most widely used type of insecticides. We examined multi-site, multi-ethnic cohort of midlife women in the U.S. that offers a unique opportunity to evaluate major determinants of OP exposure. We improved OP metabolite detection rates and obtained accurate concentrations using an improved analytical technique. Our findings suggest that consumptions of fruit, meat and alcohol are important determinants of OP exposure for midlife women. Higher concentrations of diethyl OP metabolites in Black women compared to White women, even after accounting for dietary intake, suggests additional, but unknown racial-ethnic differences that affect exposure.

Enhanced LC-ESI-MS/MS Sensitivity by Cationic Derivatization of Organophosphorus Acids.

The chemical derivatization to enhance the signal intensity and signal-to-noise (S/N) of several organophosphorus (OP) acids in liquid chromatography tandem mass spectrometry (LC-ESI-MS/MS) is illustrated. The OP class of compounds represents the environmental degradants of OP nerve agents and pesticides. N-(2-(bromomethyl)benzyl)-N,N-diethylethanaminium bromide (CAX-B) was utilized to derivatize a panel of eight acids consisting of five alkyl methylphosphonic acids (ethyl-, isopropyl-, isobutyl-, cyclohexyl-, and pinacolyl-methylphosphonic acid) along with three dialkylphosphate analogs (diethyl-, dibutyl-, and diethyl thio-phosphate). The derivatization reaction with CAX-B was conducted in acetonitrile in the presence of potassium carbonate at 70 °C for 1 h. The resulting acid derivatives were analyzed with an LC-Orbitrap-ESI-MS/MS, and their dissociation processes were investigated. It was found that the derivatization procedure increased the limits of identification (LOIs) by one to over two orders of magnitude from the range of 1 to 10 ng/mL for the intact OP-acids to the range of 0.02-0.2 ng/mL for the derivatized acids utilizing an LC-MS(QqQ) in MRM mode, regardless of the sample matrix (hair, concrete, or plant extracts). The interpretation of the corresponding ESI-MS/MS spectra for each type of derivatized sub-OP family revealed the formation of characteristic neutral losses and a characteristic ion for the organophosphorus core. This derivatization is beneficial and useful for screening and identifying target and "unknown" OP acids.

Development and Validation of a New UFLC-MS/MS Method for the Detection of Organophosphate Pesticide Metabolites in Urine.

To monitor human exposure to pesticides, experts commonly measure their metabolites in urine, particularly dialkyl phosphates (DAPs), which include diethyl phosphate (DEP), Diethyl thiophosphate (DETP), diethyl dithiophosphate (DEDTP), dimethyl phosphate (DMP), dimethyl thiophosphate (DMTP) and dimethyl dithiophosphate (DMDTP)to monitor the metabolites of organophosphates. These DAP metabolites are a urinary biomarker for assessing pesticide exposure and potential health risks. This study presented a new screening method combining ultrafast liquid chromatography with tandem mass spectrometry (UFLC-MS/MS) to detect six DAP metabolites in human urine. The study also compared standard sample extraction methods, namely, liquid-liquid extraction (LLE); quick, easy, cheap, effective, ruggedand safe (QuEChERS); and lyophilization. After a comprehensive analysis of the methods used to extract the analytes, including recovery rate, repeatability and reproducibility, the liquid-liquid extraction (LLE) method was found to be the best. It had a high recovery rate, was easy to handle, required less sample volume and had a short extraction time. Therefore, the LLE method was chosen for further analysis. The results showed excellent performance with high recoveries between 93% and 102%, precise repeatability (RSD) between 0.62% and 5.46% and acceptable reproducibility values (RSD) between 0.80% and 11.33%. The method also had limits of detection (LOD) ranging from 0.0201 ng/mL to 0.0697 ng/mL and limits of quantification (LOQ) ranging from 0.0609 ng/mL to 0.2112 ng/mL. Furthermore, the UFLC-MS/MS method was validated based on the SANTE guidance and successfully analyzed 150 urine samples from farmers and non-farmers. This validated method proved useful for biomonitoring studies focusing on OP pesticide exposure. It offers several advantages, such as a reduced need for samples, chemicals and materials, and a shorter analysis time. The method is sensitive and selective in detecting metabolites in human urine, making it a valuable approach for the practical and efficient assessment of pesticide exposure.

Exposure to non-persistent pesticides and sexual maturation of Spanish adolescent males

BackgroundSeveral non-persistent pesticides are endocrine disrupting chemicals and may impact on sexual maturation. ObjectiveTo examine the association between urinary biomarkers of non-persistent pesticides and sexual maturation in adolescent males in the Environment and Childhood (INMA) Project. MethodsThe metabolites of several pesticides were measured in spot urine samples collected from 201 boys aged 14–17 years, including: 3,5,6-trichloro-2-pyridinol (TCPy), metabolite of chlorpyrifos; 2-isopropyl-4-methyl-6-hydroxypyrimidine (IMPy), metabolite of diazinon; malathion diacid (MDA), metabolite of malathion; diethyl thiophosphate (DETP) and diethyl dithiophosphate, non-specific metabolites of organophosphates; 3-phenoxybenzoic acid (3-PBA) and dimethyl cyclopropane carboxylic acid, metabolites of pyrethroids; 1-naphthol (1-NPL), metabolite of carbaryl; and ethylene thiourea (ETU), metabolite of dithiocarbamate fungicides. Sexual maturation was assessed using Tanner stages, self-reported Pubertal Development Scale, and testicular volume (TV). Multivariate logistic regression was employed to examine associations between urinary pesticide metabolites and the odds of being in Tanner stage 5 of genital development (G5) or pubic hair growth (PH5); stage ≥4 of overall pubertal development, gonadarche, and adrenarche; or having mature TV (≥25 mL). ResultsDETP concentrations>75th percentile (P75) were associated with lower odds of being in stage G5 (OR = 0.27; 95% CI = 0.10–0.70), detectable TCPy with lower odds of gonadal stage≥4 (OR = 0.50; 95% CI = 0.26–0.96), and intermediate detectable MDA concentrations (<P75) with lower odds of adrenal stage≥4 (OR = 0.32; 95% CI = 0.11–0.94). Conversely, detectable concentrations of 1-NPL were associated with higher odds of adrenal stage≥4 (OR = 2.61; 95% CI = 1.30–5.24) but lower odds of mature TV (OR = 0.42; 95% CI = 0.19–0.90). ConclusionExposure to certain pesticides may be associated with delayed sexual maturity in adolescent males.

Associations between prenatal organophosphate pesticide exposure and placental gene networks

BackgroundExposure to organophosphate (OP) pesticides during pregnancy has been linked to deficiencies of neurobehavioral development in childhood; however, the molecular mechanisms underlying this association remain elusive. The placenta plays a crucial role in protecting the fetus from environmental insults and safeguarding proper fetal development including neurodevelopment. The aim of our study is to evaluate changes in the placental transcriptome associated with prenatal OP exposure. MethodsPregnant farm workers from two agricultural districts in northern Thailand were recruited for the Study of Asian Women and Offspring's Development and Environmental Exposures (SAWASDEE) from 2017 to 2019. For 254 participants, we measured maternal urinary concentrations of six nonspecific dialkyl phosphates (DAP) metabolites in early, middle, and late pregnancy. In parallel, we profiled the term placental transcriptome from the same participants using RNA-Sequencing and performed Weighted Gene co-expression Network Analysis (WGCNA). Generalized linear regression modeling was used to examine associations of urinary OP metabolites and placental co-expression module eigenvalues. ResultsWe identified 21 gene co-expression modules in the placenta. From the six DAP metabolites assayed, diethylphosphate (DEP) and diethylthiophosphate (DETP) were detected in more than 70% of the urine samples. Significant associations between DEP at multiple time points and two specific placental gene modules were observed. The ‘black’ module, enriched in genes involved in epithelial-to-mesenchymal transition (EMT) and hypoxia, was negatively associated with DEP in early (p = 0.034), and late pregnancies (p = 0.016). The ‘lightgreen’ module, enriched in genes involved in myogenesis and EMT, was negatively associated with DEP in late pregnancy (p = 0.010). We observed 2 hub genes (CELSR1 and PYCR1) of the ‘black’ module to be negatively associated with DEP in early and late pregnancies. ConclusionsOur results suggest that prenatal OP exposure may disrupt placental gene networks in a time-dependent manner. Such transcriptomic effects may lead to down-stream changes in placental function that ultimately affect the developing fetus.

Organophosphate pesticide exposure and biomarkers of liver injury/liver function.

There is little epidemiological evidence linking the exposure of organophosphate pesticides (OPs) to liver function or liver injury in the general population. We used data from the National Health and Nutrition Examination Survey 1999-2012 to investigate the relationship of urinary OPs with biomarkers of liver function/liver injury. The exposures were the concentrations of urinary OP metabolites (dimethyl phosphate [DMP], dimethyl thiophosphate [DMTP], diethyl phosphate [DEP] and diethyl thiophosphate [DETP]). The health outcomes were biomarkers of liver function/liver injury. The multivariable linear regression model, restricted cubic splines (RCSs) analysis and weighted quantile sum (WQS) regression were used to evaluate the relationship between individual or overall exposure of OPs and outcomes. Regressions of RCSs suggested linear and positive associations of OP metabolites with aspartate aminotransferase/alanine aminotransferase (AST/ALT) ratio (DMP and DEP) and fibrosis-4 (FIB-4) index (DMP, DEP and DMTP) (all p-non-linear values >.05). However, L-shaped relationships were found between OP metabolites (DMTP and DETP) and blood albumin and total protein (TP) concentrations (both p and non-linear values <.05). The positive associations of urinary DMP, DEP and DMTP with AST/ALT ratio, and with FIB-4 score were more pronounced among non-smokers than smokers, among alcohol drinkers than non-drinkers and among those with a body mass index (BMI) of ≥25 than participants with a BMI of <25. However, most of the interaction p values were more than .05, indicating no significant interactions between covariates and OPs on outcomes mainly including AST/ALT, FIB-4, ALB and TP levels. Finally, the WQS indices were positively associated with AST/ALT ratio (p=.014) and FIB-4 score (p=.002). Our study added novel evidence that exposures to OPs might be adversely associated with the biomarkers of liver function/liver injury. These findings indicated the potential toxic effect of OP exposures on the human liver.

Chlopyrifos fatal intoxication: homicide or suicide?

To document a fatal case where the organophosphorus compound chlorpyrifos is the cause of death, but the manner of exposure (voluntary or surreptitious) remains unclear. Chlorpyrifos is an organophosphorus insecticide used against a wide range of pests in commercially important crops and also for ornamental garden plants. In addition to being a particularly harmful substance at an environmental level, it also has a dangerous profile for humans. Voluntary suicidal intoxication generally occurs by oral ingestion and gastrointestinal absorption; systemic effects occur within minutes of inhalation when exposed to vapors or aerosols, but are delayed in the case of gastrointestinal absorption. A case of human intoxication is discussed: a 77 y.o. man was admitted at the emergency department due to a sudden alteration of the sensory, with tremors in the upper limbs and poor contactability. His GCS was 10, and he exhibited neurological symptoms of unclear etiology, with myoclonus, diffuse fasciculations, strong asthenia, hyponatremia. Family members did not refer any suspect ingestion nor peculiar circumstances. Only nine days after hospitalization, upon the release of liquid faeces mixed with sand, the toxicological evaluation of biological samples was undertaken thus disclosing the presence of chlorpyrifos. Death occurred after 30 days of coma. A judiciary autopsy was requested by the prosecutor office. Blood, stool, hair were collected during hospitalization. At the autopsy, blood from femoral vein, vitreous, urine, bile, liver, brain, fat and lung tissues were collected. A specific screening and confirmation method for pesticides was performed by GC-MS/MS at the Laboratory of Forensic Toxicology after liquid/liquid extraction with dichloromethane. Chlorpyrifos metabolites (chlorpyrifos oxon, 3,5,6-trichloro-2-pyridinol (TCPy), diethyl thiophosphate, diethylphosphate and glucuronide and sulfate conjugates of TCPy) were also investigated. In vivo: chlorpyrifos levels in blood samples were 893 ng/mL (admittance), 319 ng/mL (2 days later), 92 ng/mL (10 days later), 26 ng/mL (20 days later), and negative (<1 ng/mL) 22 days later. Metabolites were also detected but not quantified due to the lack of reference standards. In the 0-1 cm proximal section of 3 cm long white hair, the parent molecule was absent while O,O,O-triethylthiophosphate was identified. The solid material found in faeces with the appearance of sand was identified as sepiolite. Post-mortem: chlorpyrifos was detected at low levels (<1 ng/g) in fat and brain specimens; nor chlorpyrifos nor its metabolites were found in blood, urine, vitreous, bile, liver and lung specimens. A sealed bottle of “universal insecticide” found near his garden proved to be an emulsion containing chlorpyrifos, with different co-formulants but without any solid material such as sepiolite. The way of ingestion was deemed to be oral comparing the case blood levels with those reported for different ways of exposure. The doubt arose about the intentionality of the ingestion. The sepiolite found in faeces suggested the use of a solid formulation of chlorpyriphos, whereas finding the sealed bottle of liquid formulation seemed to be purportedly confusing. In insecticides containing chlorpyrifos found on the market, solid silica can be used as formulants or co-formulants, and the resulting products are described as free of significant odor, which would make them not excessively disgusting to ingest or to be administered orally as such or mixed with foods/drinks. The cause of death was acute chlorpyrifos intoxication, associated with prolonged coma, multi-organ failure and terminal cardiac arrest. There were no circumstantial and documentary elements useful for defining the dynamics of the intoxicating event as a suicide or a homicide. Voluntary suicide with organophosphorus compounds represents a common occurrence in Asian countries and in Eastern European countries but not in Italy.

Organophosphate esters exposure in relation to glucose homeostasis and type 2 diabetes in adults: A national cross-sectional study from the national health and nutrition survey

BackgroundExposure to organophosphate esters (OPEs) may be associated with impaired glucose homeostasis and increased risk of type 2 diabetes (T2D) in adolescent. However, the evidence in general population is scarce, especially for glucose homeostasis. We used data from the National Health and Nutrition Examination Survey (NHANES) 1999–2008 and 2011–2012 to investigate the relationship of urinary OPEs with glucose homeostasis and T2D in adults. MethodsA total of 5347 participants aged ≥20 years were included. The exposures were the concentrations of urinary OPEs metabolites [dimethyl phosphate (DMP), dimethyl thiophosphate (DMTP), dimethyl dithiophosphate (DMDTP), diethyl phosphate (DEP), diethyl thiophosphate (DETP), and diethyl dithiophosphate (DEDTP)]. The health outcomes were prevalence of T2D and glucose homeostasis [i.e., fasting glucose, 2-h plasma glucose during a 75-g oral glucose tolerance test (2 h-OGTT), serum insulin, HemoglobinA1c (HbA1c), HOMA-IR, and HOMA-β]. The multivariable linear regression model was used to evaluate the relationship between OPEs and glucose homeostasis. The multivariable binary logistic regression analysis was used to investigate the relationship between OPEs and prevalence of T2D. ResultsThe OPEs compound DMTP was significantly associated with higher levels of serum insulin [β (95%confidence interval, CI) = 0.21 (0.06,0.36), for one unit increase in log2-transformed exposure] and HOMA IR [β (95%CI) = 0.08 (0.02,0.14)], and increased odds of T2D [odds ratio (95% CI) = 1.05 (1.01–1.08)]. Other OPEs were not statistically associated with the serum markers for glucose homeostasis or T2D prevalence. ConclusionOur study found that the OPEs compound DMTP might be associated with impaired glucose homeostasis and may increase the prevalence of T2D in U.S. adults. Further longitudinal or experimental studies are warranted to verify our findings in different populations and different OPEs concentrations.

Exposure to non-persistent pesticides, BDNF, and behavioral function in adolescent males: Exploring a novel effect biomarker approach

BackgroundNumerous contemporary non-persistent pesticides may elicit neurodevelopmental impairments. Brain-derived neurotrophic factor (BDNF) has been proposed as a novel effect biomarker of neurological function that could help to understand the biological responses of some environmental exposures. ObjectivesTo investigate the relationship between exposure to various non-persistent pesticides, BDNF, and behavioral functioning among adolescents. MethodsThe concentrations of organophosphate (OP) insecticide metabolites 3,5,6-trichloro-2-pyridinol (TCPy), 2-isopropyl-4-methyl-6-hydroxypyrimidine (IMPy), malathion diacid (MDA), and diethyl thiophosphate (DETP); metabolites of pyrethroids 3-phenoxybenzoic acid (3-PBA) and dimethylcyclopropane carboxylic acid (DCCA), the metabolite of insecticide carbaryl 1-naphthol (1-N), and the metabolite of ethylene-bis-dithiocarbamate fungicides ethylene thiourea (ETU) were measured in spot urine samples, as well as serum BDNF protein levels and blood DNA methylation of Exon IV of BDNF gene in 15–17-year-old boys from the INMA-Granada cohort in Spain. Adolescents’ behavior was reported by parents using the Child Behavior Check List (CBCL/6–18). This study included 140 adolescents of whom 118 had data on BDNF gene DNA methylation. Multivariable linear regression, weighted quantile sum (WQS) for mixture effects, and mediation models were fit. ResultsIMPy, MDA, DCCA, and ETU were detected in more than 70% of urine samples, DETP in 53%, and TCPy, 3-PBA, and 1-N in less than 50% of samples. Higher levels of IMPy, TCPy, and ETU were significantly associated with more behavioral problems as social, thought problems, and rule-breaking symptoms. IMPy, MDA, DETP, and 1-N were significantly associated with decreased serum BDNF levels, while MDA, 3-PBA, and ETU were associated with higher DNA methylation percentages at several CpGs. WQS models suggest a mixture effect on more behavioral problems and BDNF DNA methylation at several CpGs. A mediated effect of serum BDNF within IMPy-thought and IMPy-rule breaking associations was suggested. ConclusionBDNF biomarkers measured at different levels of biological complexity provided novel information regarding the potential disruption of behavioral function due to contemporary pesticides, highlighting exposure to diazinon (IMPy) and the combined effect of IMPy, MDA, DCCA, and ETU. However, further research is warranted.

Urinary organophosphate metabolite concentrations and birth sizes among women conceiving through in vitro fertilization in Shanghai, China

BackgroundFew studies have examined the adverse birth sizes of preconception exposure to organophosphate pesticides (OPs) in women undergoing in vitro fertilization (IVF). ObjectivesWe investigated the relationship of preconception OP exposure with birth sizes among Chinese women undergoing IVF. MethodsThis study included 302 couples seeking infertility treatment in the China National Birth Cohort Study, from Shanghai, China, who gave birth to singleton infants between 2018 and 2021. Clinical data were collected from medical records. We measured the concentrations of six nonspecific dialkyl phosphates (DAP) metabolites of OPs [diethylthiophosphate (DETP), diethylphosphate (DEP), diethyldithiophosphate (DEDTP), dimethyldithiophosphate (DMTP), dimethylphosphate (DMP), and dimethyldithiophosphate (DMDTP)] in maternal urine. DMDTP and DEDTP were precluded from further analyses due to the low detection rates. Generalized linear models (GLMs) and weighted quantile sum (WQS) regression analyses were performed to examine the individual and joint effects of OP exposures on gestational age, birth weight, body length, and ponderal index. Odder ratio (OR) of preterm birth were estimated using logistic regression models. ResultsWomen in the highest as compared with lowest quartile of DEP had shorter gestational age (β = - 0.68; 95% CI = −1.24, −0.11). The association was modified by sex, with boys showing larger decreases in gestational age (β = - 0.86; 95% CI = −1.60, −0.13). No associations were found between other DAP metabolites and birth sizes. Results from linear models with individual DAP metabolites were corroborated by the WQS regression where DEP had the largest contribution to the overall mixture effect on gestational age (weight = 0.70). Moreover, DEP concentration was associated with an elevated risk of preterm birth (OR = 1.35, 95% CI = 1.11, 2.25). ConclusionPreconception DEP concentration was associated with shortened gestational age and increased risk of preterm birth, and the association was more pronounced among boys than girls.

Association between urinary dialkylphosphate metabolites and dyslexia among children from three cities of China: The READ program

Exposure to organophosphate (OP) insecticides has been found to be related to neurodevelopmental disorders in children. However, no study has examined the association between OP insecticide exposure and the risk of dyslexia among children. We aimed to explore the association between OP insecticide exposure, indicated by urinary dialkylphosphate metabolites (DAPs), and the risk of dyslexia among Chinese Han children from three cities. A total of 845 children (422 dyslexics and 423 non-dyslexics) from Tongji Reading Environment and Dyslexia research program were included in the current case-control study. We measured six DAPs in urine samples, collected from November 2017 to December 2020. Logistic regression models were used to estimate odds ratios (ORs) for the association between DAPs and dyslexia risk, adjusting for potential confounders. The detection frequencies of DAPs were above 97.5%, except for diethyldithiophosphate and dimethyldithiophosphate. Diethyl phosphate metabolites (DEs) were significantly associated with the risk of dyslexia. Compared with the lowest quartile, the adjusted ORs of dyslexia risk for the highest quartile of urinary diethylthiophosphate (DETP) and diethylphosphate (DEP) were 1.82 (1.04, 3.20) and 1.85 (1.08, 3.17), respectively. In addition, the adjusted ORs for dyslexia per 10-fold of urinary DEP, DETP, and ∑DEs concentration were 1.87 (1.12, 3.13), 1.55 (1.03, 2.35), and 1.91 (1.13, 3.21), respectively. Analyses stratified by gender indicated that such associations were more significant among boys. This study suggested that exposure to OP insecticides may be related to dyslexia among Chinese Han children from the three studied cities. However, our results should be interpreted with caution because of the case-control design and the fact that only one-spot urine sample was collected from the children. More studies with children living in China are necessary concerning the relatively high levels of urinary OP metabolites in our study.

Ecological safety with multifunctional applications of biogenic mono and bimetallic (Au–Ag) alloy nanoparticles

Recently, the design and biosynthesis of metallic nanoparticles (NPs) have drawn immense interest, but their very specific function and secondary toxic effects are major concern towards commercial application of NPs. That's why environment-friendly (nontoxic) NPs having multiple functions are extremely important. Herein, we report the mechanism of biosynthesis of mono and bimetallic (Au–Ag) alloy NPs and study their multifunctional (antioxidant, antifungal and catalytic) activity and ecotoxicological property. AgNPs exhibit phytotoxicity (at 100 μg/ml) on morphological characteristics of Lentil (during germination), while alloy and AuNPs are non-toxic (up to 100 μg/ml). In-vitro antioxidant response using DPPH methods reveals that alloy NPs (IC50 = 55.8 μg/ml) possesses better antioxidant activity compared to the monometallic NPs (IC50 = 73.6–82.6 μg/ml). In addition, alloy NPs displayed appreciable antifungal efficacy against a plant pathogenic fungus Gloeosporium musarum by structural damage to hyphae and conidia of the fungus. The catalytic performance of NPs for degradation of chlorpyriphos (CP) pesticide reveals that alloy NPs is more efficient in terms of rate constant (k = 0.405 d−1) and half-life (T50 = 1.71 d) compared to the monometallic counterparts (k = 0.115–0.178 d−1; T50 = 3.89–6.04 d). Degradation products of CP (3,5,6-trichloropyridinol and diethyl thiophosphate) are confirmed using mass spectrometry and based on that a degradation pathway has been suggested. Thus, these sustainable and ecological safe biogenic (Au-Ag) alloy NPs promise multiple applications as an antioxidant in the pharmaceutical sector, as a fungicide for disease control in agriculture, as a catalyst for remediation of toxic pollutants and in other pertinent areas.

Chlorpyrifos Disrupts Acetylcholine Metabolism Across Model Blood-Brain Barrier.

Despite the significant progress in both scientific understanding and regulations, the safety of agricultural pesticides continues to be called into question. The need for complementary analytics to identify dysregulation events associated with chemical exposure and leverage this information to predict biological responses remains. Here, we present a platform that combines a model organ-on-chip neurovascular unit (NVU) with targeted mass spectrometry (MS) and electrochemical analysis to assess the impact of organophosphate (OP) exposure on blood-brain barrier (BBB) function. Using the NVU to simulate exposure, an escalating dose of the organophosphate chlorpyrifos (CPF) was administered. With up to 10 μM, neither CPF nor its metabolites were detected across the BBB (limit of quantitation 0.1 µM). At 30 µM CPF and above, targeted MS detected the main urinary metabolite, trichloropyridinol (TCP), across the BBB (0.025 µM) and no other metabolites. In the vascular chamber where CPF was directly applied, two primary metabolites of CPF, TCP and diethylthiophosphate (DETP), were both detected (0.1–5.7 µM). In a second experiment, a constant dose of 10 µM CPF was administered to the NVU, and though neither CPF nor its metabolites were detected across the BBB after 24 h, electrochemical analysis detected increases in acetylcholine levels on both sides of the BBB (up to 24.8 ± 3.4 µM) and these levels remained high over the course of treatment. In the vascular chamber where CPF was directly applied, only TCP was detected (ranging from 0.06 μM at 2 h to 0.19 μM at 24 h). These results provide chemical evidence of the substantial disruption induced by this widely used commercial pesticide. This work reinforces previously observed OP metabolism and mechanisms of impact, validates the use of the NVU for OP toxicology testing, and provides a model platform for analyzing these organotypic systems.

Biosensors for the detection of organophosphate exposure by a new diethyl thiophosphate-specific aptamer.

An aptamer specifically binding to diethyl thiophosphate (DETP) was constructed and incorporated in an optical sensor and electrochemical techniques to enable the specific measurement of DETP as a metabolite and a biomarker of organophosphate exposure. A DETP-bound aptamer was selected from the library using capillary electrophoresis-systematic evolution of ligands by exponential enrichment (CE-SELEX). A colorimetric method revealed that the aptamer had the highest affinity for DETP, with a mean Kd value (± SD) of 0.103 ± 0.014µM. The docking results and changes in resistance showed that the selectivity of the aptamer for DETP was higher than that for the similar structures of dithiophosphate (DEDTP) and diethyl phosphate (DEP). The altered amplitude of cyclic voltammetry showed a linear range of DETP detection covering 0.0001-10µg/ml with a limit of detection of 0.007µg/ml. The recovery value of a real sample of pH 7 was 97.2%. The current method showed great promise in using the DETP-specific aptamer to detect the exposure history to organophosphates by measuring their metabolites, although degradation of organophosphate parent compounds might occur.

Application of Q-TOF–MS based metabonomics techniques to analyze the plasma metabolic profile changes on rats following death due to acute intoxication of phorate

Organophosphorus pesticides (OPS) are widely used in the world, and many poisoning cases were caused by them. Phorate intoxication is especially common in China. However, there are currently few methods for discriminating phorate poisoning death from phorate exposure after death and interpretation of false-positive results due to the lack of effective biomarkers. In this study, we investigated the metabonomics of rat plasma at different dose levels of acute phorate intoxication using ultra-performance liquid chromatography quadrupole-time of flight mass spectrometry (UPLC-Q-TOF-MS) analysis. A total of 11 endogenous metabolites were significantly changed in the groups exposed to phorate at LD50 level and three times of LD50 (3LD50) level compared with the control group, which could be potential biomarkers of acute phorate intoxication. Plasma metabonomics analysis showed that diethylthiophosphate (DETP) could be a useful biomarker of acute phorate intoxication. The levels of uric acid, acylcarnitine, succinate, gluconic acid, and phosphatidylcholine (PC) (36:2) were increased, while pyruvate level was decreased in all groups exposed to phorate. The levels of ceramides (Cer) (d 18:0/16:0), palmitic acid, and lysophosphatidylcholine (lysoPC) (18:1) were only changed after 3LD50 dosage. The results of this study indicate that the dose-dependent relationship exists between metabolomic profile change and toxicities associated with apoptosis, fatty acid metabolism disorder, energy metabolism disorder especially tricarboxylic acid (TCA) cycle, as well as liver, kidney, and nervous system functions after acute exposure of phorate. This study shows that metabonomics is a useful tool in identifying biomarkers for the forensic toxicology study of phorate poisoning.

Organophosphate Pesticide Exposures in Early and Late Pregnancy Influence Different Aspects of Infant Developmental Performance

Organophosphate (OP) pesticides can transfer from mother to fetus via the placenta and amniotic fluid and may affect the development of infants. This study aims to evaluate the associations between maternal OP concentrations collected in the 1st–2nd trimester and the 3rd trimester of pregnancy and the infant developmental performance. The Screening Test of the Bayley Scales of Infants and Toddler Development, Third Edition (BSID–III screening test) was used to assess development performance at 2 and 6 months of age. Multiple regression analysis showed a negative correlation between cognitive performance at 2 months and maternal diethylthiophosphate (DETP) levels in the 1st–2nd trimester (β ± SE = −0.012 ± 0.004, p < 0.05). We also found that expressive communication and fine motor performance at 6 months were negatively associated with maternal diethyldithiophosphate (DEDTP) levels in the 3rd trimester (β ± SE = −0.047 ± 0.016, p < 0.05, and β ± SE = −0.044 ± 0.017, p < 0.05, respectively). These results suggest that maternal ethylated OP concentrations at different timing of exposure during pregnancy may influence different aspects of infant developmental performance.

Aptasensor for paraquat detection by gold nanoparticle colorimetric method

This study aimed to develop an aptasensor for paraquat detection by gold nanoparticles. The specific aptamer for paraquat was selected by using the SELEX process via capillary electrophoresis. Sixty-three aptamer candidates were amplified by asymmetric PCR and screened for paraquat binding using gold nanoparticles (AuNPs). Aggregation of AuNPs was specifically induced by desorption of paraquat binding aptamers from the surface of AuNPs as a result of aptamer–target interaction leading to the color change from red to purple. Aptamer 77F with the following sequence: 5′-AGGCTTACACCTGAAAAGCGGCTTAATTTACACTACTGTAT-3′ was selected as a highly specific aptamer for paraquat. The detection limit of paraquat was 0.267 µg/mL by colorimetry and 1.573 µg/mL by the quartz crystal microbalance (QCM) technique. This aptamer showed specificity for paraquat by colorimetry. Dimethyl phophite, diethyl phophite and O,O diethyl thiophosphate potassium salt did not react by colorimetry but, exhibited a weak nonspecific reaction by QCM. This is first time that an aptasensor was used for detection of paraquat based on QCM. However, the aptasensor based on the colorimetric method simply uses a generation of a signal that can be detected by the naked eye. This technique is rapid, low cost easy to use and suitable for on-site detection of herbicide residue.

Organophosphate pesticide exposure, hormone levels, and interaction with PON1 polymorphisms in male adolescents

ObjectiveTo examine the association between urinary metabolites of organophosphate (OP) pesticides and serum concentrations of thyroid and reproductive hormones in male adolescents and to assess the potential effect of interactions between OP pesticides and paraoxonase 1 (PON1) polymorphisms on hormone levels. MethodsStudy subjects (N = 117) were male 16- to 17-year-olds from the Environment and Childhood (INMA)-Granada cohort in Spain. Concentrations of 3,5,6-trichloro-2-pyridinol (TCPy), a metabolite of chlorpyrifos/chlorpyrifos-methyl, 2-isopropyl-6-methyl-4-pyrimidinol (IMPy), a metabolite of diazinon, and diethylthiophosphate (DETP) and diethyldithiophosphate (DEDTP), non-specific metabolites of OP pesticides, were measured in a spot urine sample from each subject and adjusted for creatinine. Levels of reproductive hormones (total testosterone [TT], estradiol [E2], dehydroepiandrosterone sulfate [DHEAS], sex hormone binding globulin [SHBG], luteinizing hormone [LH], follicle stimulating hormone [FSH], anti-Müllerian hormone [AMH], insulin growth factor 1 [IGF-1], and prolactin), thyroid hormones (free thyroxine [FT4], total triiodothyronine [TT3], and thyroid stimulating hormone [TSH]), and PON1 Q192R and L55M polymorphisms were determined in blood drawn during the same clinical visit. ResultsMultiple linear regression models showed that detectable levels of TCPy were associated with an increase in DHEAS and decreases in E2, FSH, and AMH; detectable IMPy with increases in E2, DHEAS, FSH, AMH, and prolactin and decreases in SHBG and LH; and detectable DETP with marginally-significant increases in TT and TT3 and decreases in FSH, AMH, and prolactin. The effect of IMPy and DETP on DHEAS and TT levels, respectively, was higher in subjects that carried the PON1 55MM genotype, while the effect of TCPy, IMPy, and DETP on thyroid hormone levels was higher in PON1 192QR/RR or 55MM genotype carriers. ConclusionsIn male adolescents, non-occupational exposure to OP pesticides was associated with several changes in reproductive and thyroid hormone levels, and the magnitude of some associations was greater in adolescents genetically more susceptible to OP pesticide exposure who carry the PON1 55MM genotype.