利用两份口服草铵膦中毒死者提取的心血样本,分析了人体内草铵膦代谢物的存在情况,明确了草铵膦中毒死者血液的检测目标物为草铵膦及其主要代谢物3-甲基膦基-丙酸,并提出了高效液相色谱法-串联质谱法测定血液中草铵膦及其代谢物3-甲基膦基-丙酸含量的方法。在血液、水、乙腈的体积比为1∶0.5∶3条件下沉淀蛋白,获取上清液后添加与乙腈体积相同的二氯甲烷,涡旋、离心,上层水相经0.22 μm水相滤膜过滤,所得滤液注入高效液相色谱仪。以价格低廉的Dikma Polyamino HILIC色谱柱(150 mm×2.0 mm,5 μm)为固定相,以含5 mmol·L^-1乙酸铵的1.25%(体积分数)氨水溶液-乙腈为流动相体系进行梯度洗脱;分离后的目标物经电喷雾离子源负离子模式扫描,多反应监测模式检测,以基质匹配法定量。结果显示:草铵膦和3-甲基膦基-丙酸工作曲线的线性范围均为0.2~10.0 mg·L^-1,检出限(3S/N)均为0.1 mg·L^-1;对空白心血进行3个浓度水平的加标回收试验,草铵膦和3-甲基膦基-丙酸的回收率分别为35.6%~50.1%和50.6%~70.1%,日内精密度(n=6)分别为5.7%~8.7%和6.9%~8.9%,日间精密度(n=6)分别为5.7%~6.3%和4.2%~9.6%;对两份实际案例的血液检材进行定量分析,其中草铵膦质量浓度为5.8 mg·L^-1和7.5 mg·L^-1,代谢物3-甲基膦基-丙酸质量浓度为5.3 mg·L^-1和5.8 mg·L^-1。

Two heart blood samples of poisoning decedents caused by oral glufosinate-ammonium were used to analyze the presence of glufosinate-ammonium metabolites in human body, and it was determined that the detection targets in blood of poisoning deceden caused by glufosinate-ammonium were glufosinate-ammonium and its main metabolite 3-methylphosphinico-propionic acid, and a method for the determination of glufosinate-ammonium and its metabolite 3-methylphosphinico-propionic acid in blood by high performance liquid chromatography-tandem mass spectrometry was proposed. The protein was precipitated under the condition of blood, water and acetonitrile at volume ratio of 1∶0.5∶3, and the obtained supernatant was mixed with dichloromethane with the same volume as acetonitrile. After vortex and centrifugation, the upper water phase was filtered by 0.22 μm water phase filter membrane, and the resulting filtrate was injected into high performance liquid chromatograph. Gradient elution was performed on a cheap Dikma Polyamino HILIC column (150 mm×2.0 mm, 5 μm) as stationary phase, with 1.25% (volume fraction) ammonia solution containing 5 mmol·L^-1 ammonium acetate-acetonitrile as mobile phase system. The separated targets were scanned by electrospray ion source with negative ion mode, and detected by multi-reaction monitoring mode, with matrix matching method for quantification. As shown by the results, the linear ranges of the working curves for glufosinate-ammonium and 3-methylphosphinico-propionic acid were 0.2-10.0 mg·L^-1, with detection limits (3S/N) of 0.1 mg·L^-1. Test for recovery was made on the blank heart blood at 3 concentration levels by standard addition method, giving results of glufosinate-ammonium and 3-methylphosphinico-propionic acid in the ranges of 35.6%-50.1% and 50.6%-70.1%, respectively,with intra-day precision (n=6) in the ranges of 5.7%-8.7% and 6.9%-8.9%, respectively, and inter-day precision (n=6) in the ranges of 5.7%-6.3% and 4.2%-9.6%, respectively. The two blood samples from actual cases were quantitatively analyzed, in which the mass concentrations of glufosinate-ammonium were 5.8 mg·L^-1 and 7.5 mg·L^-1, and the mass concentrations of metabolite 3-methylphosphinico-propionic acid were 5.3 mg·L^-1 and 5.8 mg·L^-1.