Department of Legal Medicine and Bioethics, Nagoya University Graduate School of Medicine, Nagoya, Japan.
Nagoya J Med Sci. 2021 Aug;83(3):567-587. doi: 10.18999/nagjms.83.3.567.
Glyphosate (GLYP) and glufosinate (GLUF) are phosphorus-containing amino acid type herbicides that are used worldwide. With their rising consumptions, fatal intoxication cases due to these herbicides, whether accidental or intentional, cannot be ignored. Both compounds are difficult to detect, and their pretreatment for instrumental analysis are complicated and time-consuming. Our aim was to develop a simple and rapid quantification method for the two herbicides and their metabolites with liquid chromatography/tandem mass spectrometry (LC/MS/MS). We also compared 2-amino-4-phosphonobutyric acid and DL-2-amino-5-phosphonopentanoic acid as alternative internal standards (IS) to GLYPC N. Herbicide-containing specimens were highly diluted, evaporated to dryness, and derivatized with acetate/acetic anhydride and trimethyl orthoacetate for 30 min. at 120°C. Our optimized LC conditions successfully separated the target analytes, with acceptable linearities ( >0.98) and matrix effects (65%-140%). Accuracy and precision ranged from 80.2 % to 111 %, and from 1.3 % to 13 % at the higher concentration, respectively.The concentration of the herbicides and their metabolites were investigated in a postmortem case of suspected herbicide poisoning cases, in which we detected GLYP and its metabolites. Using one of the three ISs, the GLYP concentrations ranged from 3.1 to 3.5 mg/mL, and 3.3 to 4.5 mg/mL in plasma and urine, respectively; GLYP metabolite concentrations in plasma and urine were 18 to 20 μg/mL and 44 to 54 μg/mL. We thus succeeded in developing a rapid method without extraction for measuring GLYP and GLUF along with their metabolites, and demonstrated its practical applicability.
草甘膦（GLYP）和草铵膦（GLUF）是含磷的氨基酸类型除草剂，在世界范围内广泛使用。随着它们的消耗量不断增加，无论是意外还是故意摄入这些除草剂导致的致命中毒病例不容忽视。这两种化合物都很难检测，而且它们的仪器分析前处理既复杂又耗时。我们的目的是开发一种简单快速的定量方法，用于同时检测这两种除草剂及其代谢物的液相色谱/串联质谱（LC/MS/MS）。我们还比较了 2-氨基-4-膦酸丁酸和 DL-2-氨基-5-膦戊酸作为替代内标（IS）与 GLYPC N。含除草剂的标本经过高度稀释、蒸发至干，然后用乙酸/乙酸酐和三甲氧基乙酸在 120°C 下衍生 30 分钟。我们优化的 LC 条件成功地分离了目标分析物，具有可接受的线性（>0.98）和基质效应（65%-140%）。准确度和精密度分别在高浓度下为 80.2%至 111%，在 1.3%至 13%之间。我们在一例疑似除草剂中毒的死后案例中研究了这些除草剂及其代谢物的浓度，在该案例中我们检测到了 GLYP 和其代谢物。使用三种 IS 中的一种，GLYP 在血浆和尿液中的浓度分别为 3.1 至 3.5 毫克/毫升和 3.3 至 4.5 毫克/毫升；GLYP 代谢物在血浆和尿液中的浓度分别为 18 至 20 微克/毫升和 44 至 54 微克/毫升。因此，我们成功开发了一种无需提取即可测量 GLYP 和 GLUF 及其代谢物的快速方法，并证明了其实际适用性。
