Abstract

Identifying the source of ethanol in a decedent remained a complicated problem for forensic toxicologists because of postmortem ethanol formation. As ethanol’s non-oxidative metabolites, ethyl glucuronide (EtG) and ethyl sulfate (EtS) have the potential to distinguish between antemortem ethanol consumption and postmortem ethanol formation, due to their high sensitivity and selectivity. In the current study, a simple and quick liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed for the determination of EtG and EtS in human whole blood and vitreous humor (VH). A total of 20 μL of the sample was precipitated by methanol, and the analytes were detected by LC-MS/MS in a run of 6 min. This method achieved high sensitivity (limits of detection: 2 ng/mL for both EtG and EtS), with linearity in the range of 5–10,000 ng/mL in both whole blood and VH. Deviations in accuracy, inter- and intra-day precision were all lower than 15% at three quality control levels. Subsequently, this method was applied to 62 real forensic cases. Only blood samples were available in 52 cases. Paired blood and VH samples were present in 10 cases. The concentrations of EtG and EtS in blood were in the range of 0–22,264.8 ng/mL and 0–2,126.0 ng/mL, respectively. In one case with both blood and VH, the blood ethanol concentration was 1.22 mg/mL, with EtG and EtS both below limits of quantification (5 ng/mL) in VH, and no EtG and EtS found in whole blood. The results suggested that EtG and EtS were useful markers for the interpretation of ethanol resource in postmortem blood and VH.

中文翻译：

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液相色谱-质谱-质谱法测定人全血和玻璃体液中的乙醛酸乙酯和硫酸乙酯及其在尸体酒精检测结果的解释中的应用

摘要

由于死后乙醇的形成，对死者中乙醇的来源进行鉴定对于法医毒理学家仍然是一个复杂的问题。作为乙醇的非氧化性代谢产物，葡糖醛酸乙酯（EtG）和硫酸乙酯（EtS）由于具有高灵敏度和选择性，因此有可能区分前验乙醇消耗和死后乙醇形成。在当前的研究中，开发了一种简单快速的液相色谱串联质谱法（LC-MS / MS），用于测定人全血和玻璃体液（VH）中的EtG和EtS。用甲醇沉淀总共20μL的样品，并在6分钟的运行中通过LC-MS / MS检测分析物。此方法实现了高灵敏度（检测极限：EtG和EtS均为2 ng / mL），线性范围为5-10，全血和VH中的浓度均为000 ng / mL。在三个质量控制级别上，准确性，日间和日内精度的偏差均低于15％。随后，该方法被应用于62个真实的法医案件。在52例中只有血液样本可用。配对的血液和VH样本有10例。血液中EtG和EtS的浓度分别为0-222.64.8 ng / mL和0-21266.0 ng / mL。在同时有血液和VH的情况下，血液乙醇浓度为1.22 mg / mL，EtG和EtS均低于VH的定量限（5 ng / mL），并且在全血中未发现EtG和EtS。结果表明，EtG和EtS是解释死后血液和VH中乙醇资源的有用标记。在三个质量控制级别上，日间和日间精度均低于15％。随后，该方法被应用于62个真实的法医案件。在52例中只有血液样本可用。配对的血液和VH样本有10例。血液中EtG和EtS的浓度分别为0-222.64.8 ng / mL和0-21266.0 ng / mL。在同时有血液和VH的情况下，血液乙醇浓度为1.22 mg / mL，EtG和EtS均低于VH的定量限（5 ng / mL），并且在全血中未发现EtG和EtS。结果表明，EtG和EtS是解释死后血液和VH中乙醇资源的有用标记。在三个质量控制级别上，日间和日间精度均低于15％。随后，该方法被应用于62个真实的法医案件。在52例中只有血液样本可用。配对的血液和VH样本有10例。血液中EtG和EtS的浓度分别为0-222.64.8 ng / mL和0-21266.0 ng / mL。在同时有血液和VH的情况下，血液乙醇浓度为1.22 mg / mL，EtG和EtS均低于VH的定量限（5 ng / mL），并且在全血中未发现EtG和EtS。结果表明，EtG和EtS是解释死后血液和VH中乙醇资源的有用标记。配对的血液和VH样本有10例。血液中EtG和EtS的浓度分别为0-222.64.8 ng / mL和0-21266.0 ng / mL。在同时有血液和VH的情况下，血液乙醇浓度为1.22 mg / mL，EtG和EtS均低于VH的定量限（5 ng / mL），并且在全血中未发现EtG和EtS。结果表明，EtG和EtS是解释死后血液和VH中乙醇资源的有用标记。配对的血液和VH样本有10例。血液中EtG和EtS的浓度分别为0-222.64.8 ng / mL和0-21266.0 ng / mL。在同时有血液和VH的情况下，血液乙醇浓度为1.22 mg / mL，EtG和EtS均低于VH的定量限（5 ng / mL），并且在全血中未发现EtG和EtS。结果表明，EtG和EtS是解释死后血液和VH中乙醇资源的有用标记。