LC-MS/MS检测血液中草铵膦、草甘膦及其代谢物

Determination of glyphosate,glufosinate-ammonium and their metabolites in blood by LC-MS/MS

目的 建立血液中草铵膦、草甘膦及其代谢物3-(甲基膦基)-丙酸、氨甲基膦酸的液相色谱-串联质谱(LC-MS/MS)检验方法。方法 对血液前处理的传统方法 -乙腈沉淀蛋白法进行改进,通过添加适量水发现有利于目标毒物从血液中释放出来,并增加在上清液中添加同乙腈比例的二氯甲烷处理步骤,实现上清液有机相与水相分层,直接吸取水层过水相滤膜供检,起到对目标毒物浓缩、净化效果;以含2 mmol/L乙酸铵1.25%氨水(v/v)水溶液和乙腈作为流动相,在DikmaPolyamino HILIC (150 mm×2.0 mm,5μm)液相色谱柱上实现良好的分离,以多反应离子负模式进行串联质谱检测。结果 血液中草铵膦、草甘膦及其代谢物-3-(甲基膦基)-丙酸、氨甲基膦酸检测限分别达到0.1μg/mL、0.4μg/mL、0.2μg/mL和0.4μg/mL,线性范围分别为0.2μg/mL~10.0μg/mL、0.6μg/m L~10.0μg/mL、0.4μg/mL~10.0μg/mL和0.6μg/mL~10.0μg/mL;日内和日间精密度RSD均小于15%。结论为快速检测血液中草铵膦、草甘膦及其代谢物提供了低成本有效的分析途径,也为探索强极性毒物的血液前处理方法提供了思路。

Objective To determine glufosinate,glyphosate and their metabolites 3-(methylphosphinico)-propionic acid and aminomethylphosphonic acid in human blood by LC-MS/MS.Methods The traditional method of blood pretreatment-acetonitrile precipitation protein method was improved.Proper amount of water was added to promote the release of target poison from blood.Dichloromethane in the same proportion as acetonitrile was added into the supernatant to separate the organic phase from the water phase.The water phase was directly absorbed by membranes to concentrate and purify the target poison.Liquid chromatography was performed on a Dikmapolino HILIC(150 mm × 2.0 mm,5μm)liquid chromatographic column using 1.25 % ammonium acetate(v/v) aqueous solution and acetonitrile as the mobile phase.Results The detection limits of glufosinate,glyphosate and their metabolites-3-(methylphosphinico)-propionic acid and aminomethylphosphonic acid in blood were 0.1μg/mL,0.4μg/mL,0.2μg/mL and 0.4μg/mL,respectively,and the linear ranges were 0.2 μg/mL ~ 10.0 μg/mL,0.6 μg/mL ~ 10.0 μg/mL and 0.4 μg/mL ~ 10.0 μg/mL and 0.6 μg/mL ~ 10.0 μg/mL,respectively.The RSD of intra-day and inter-day precision were less than 15 %.Conclusion We provide a low-cost and effective method for the rapid determination of glufosinate,glyphosate and their metabolites in blood.We also provide an idea for exploring the pretreatment method for blood containing strong polar toxicants.