Food Safety Division, ICMR-National Institute of Nutrition, Hyderabad 500007, Telangana, India.
Department of Biochemistry, Osmania University, Hyderabad 500007, Telangana, India.
Molecules. 2023 Aug 1;28(15):5800. doi: 10.3390/molecules28155800.
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.
为了监测人类接触农药的情况，专家通常会测量其尿液中的代谢物，特别是二烷基磷酸酯（DAPs），包括二乙基磷酸酯（DEP）、二乙基硫代磷酸酯（DETP）、二乙基二硫代磷酸酯（DEDTP）、磷酸二甲酯（DMP）、磷酸二硫代甲基酯（DMTP）和磷酸二硫代二甲基酯（DMDTP），以监测有机磷农药的代谢物。这些 DAP 代谢物是评估农药暴露和潜在健康风险的尿液生物标志物。本研究提出了一种新的筛选方法，结合超快速液相色谱与串联质谱（UFLC-MS/MS）检测人尿液中的六种 DAP 代谢物。该研究还比较了标准样品提取方法，即液液萃取（LLE）；快速、简单、廉价、有效、耐用和安全（QuEChERS）；和冻干法。在对包括回收率、重复性和重现性在内的分析物提取方法进行全面分析后，发现液液萃取（LLE）法效果最佳。该方法回收率高，易于操作，所需样品量少，提取时间短。因此，选择 LLE 方法进行进一步分析。结果表明，该方法具有出色的性能，回收率在 93%至 102%之间，精密度（RSD）在 0.62%至 5.46%之间，可接受的重现性（RSD）值在 0.80%至 11.33%之间。该方法的检测限（LOD）范围为 0.0201ng/mL 至 0.0697ng/mL，定量限（LOQ）范围为 0.0609ng/mL 至 0.2112ng/mL。此外，该 UFLC-MS/MS 方法是根据 SANTE 指南进行验证的，并成功分析了 150 名农民和非农民的尿液样本。该验证方法为关注有机磷农药暴露的生物监测研究提供了有用的工具。它具有减少对样品、化学物质和材料的需求以及缩短分析时间等优点。该方法在检测人尿液中的代谢物方面具有较高的灵敏度和选择性，是一种实用高效的评估农药暴露的方法。
