LC-MS/MS测定血液、肝组织中的欧夹竹桃苷

摘要

目的 建立血液和肝组织中欧夹竹桃苷的液相色谱-串联质谱法(liquid chromatography-tandem mass spectrometry, LC-MS/MS),并应用于夹竹桃中毒致死案例中.方法 采用乙醚对血液或肝组织进行液液提取,提取物经Agilent ZORBAX SB-C18柱分离,以乙腈和20 mmol/L乙酸铵(含0.1％甲酸)溶液进行梯度洗脱,采用电喷雾正离子化(electrospray positive ionization, ESI+)、多反应监测模式检测欧夹竹桃苷.结果 血液和肝组织中欧夹竹桃苷在相应的线性范围内线性良好(r＞0.995 0),检出限分别为1 ng/mL和2 ng/g,提取回收率均大于70.50％,日内和日间精密度均小于10.71％,准确度为98.42％～111.63％,基质效应为91.52％～106.39％.该方法成功应用于1例疑似夹竹桃中毒致死案例,在尸体心血、尿液、肝组织、胆汁、胃壁组织和胃内容物中均检出欧夹竹桃苷,其含量范围为65.5～29 600.0 ng/mL(ng/g).结论 本研究建立的方法操作简便、选择性好,适用于血液、肝组织等生物检材中的欧夹竹桃苷分析,可为夹竹桃中毒的法医学鉴定和临床诊治提供技术保障.%Objective To develop a liquid chromatography-tandem mass spectrometry (LC-MS/MS) analytical method for the determination of oleandrin in blood and liver tissues, which could be applied to the cases of death caused by oleander poisoning.Methods Blood or liver tissues underwent a liquidliquid extraction (LLE) using ethyl acetate, and the extract was separated on an Agilent ZORBAX SB-C18column and eluted with a gradient of acetonitrile and 20 mmol/L ammonium acetate (containing0.1％formic acid).Oleandrin was detected using electrospray positive ionization (ESI+) with multiplereaction monitoring (MRM) mode.Results Oleandrin showed excellent linearity in both blood and liver samples in the corresponding linear range (r＞0.995 0), with detection limits 1 ng/mL and 2 ng/g, respectively, extraction recovery rates greater than 70.50％, both intra-and inter-day precisions less than10.71％, accuracies 98.42％-111.63％, and matrix effects 91.52％-106.39％.The method was successfully applied to a case of suspected oleander poisoning.Oleandrin was detected in the blood, urine, liver tissues, bile, stomach wall tissues and stomach contents of the cadaver, with the content ranging from65.5 to 29 600.0 ng/mL (ng/g).Conclusion The method developed in this study is simple and convenient to operate with good selectivity, and is suitable for the analysis of oleandrin in biological samples such as blood and liver tissues, which can provide technical support for forensic identification and clinical diagnosis and treatment of oleander poisoning.