Method Consolidation to Improve Scope and Efficiency in Postmortem Toxicology
Method Consolidation to Improve Scope and Efficiency in Postmortem Toxicology
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DOI：
10.1093/jat/bkaa003
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发表时间：
2020-06-01
影响因子：
2.5
通讯作者：
Rodda, Luke N.
中科院分区：
文献类型：
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作者：
Gevorkyan, Jirair;Wong, Megan;Rodda, Luke N.
系统毒理学方法同时采用意识形态的变化以及仪器和样品提取的改进，可以通过降低灵敏度、提高特异性和最小化资源使用来提高毒理学测试效率。从历史上看，旧金山首席法医办公室严重依赖气相色谱-质谱 (GC-MS) 测试体系，其中包括单个药物类别的确认和定量测定。利用 GC-MS 的传统方法通常需要重复测试，耗尽样品量，并影响生产率和周转时间，特别是对于现代尸检毒理学中常见的多重用药案例。这里描述的方法将七种传统方法的范围整合到单一的液相色谱-串联质谱 (LC-MS-MS) 方法中，以实现更好的灵敏度、更高的通量、最小化滥用药物定量的样品消耗，并通过结合智能、自动化处理来提高质量保证。使用简单的乙腈蛋白破碎和随后的小瓶内过滤，快速提取约 100 μL 的血液或尿液，并将其注射到 LC-MS-MS 系统中。开发的方法经过了 SWGTOX 和国际指南的全面验证，并纳入了 55 种分析物以及定制查询，有助于快速、一致地应用数据处理和审查的可接受性标准。通过对 1,389 个样本（858 个血液和 531 个尿液）的分析证明了其适用性，其中至少 41% 的阳性结果可能由于灵敏度降低而被遗漏，并且 11% 的结果不在先前分析方法估计的范围内。平均而言，本研究中的病例以前需要进行 3 次不同的 GC-MS 检测、3 mL 血液以及超过 30 小时的工作人员时间。所描述的 LC-MS-MS 分析方法减少了执行多次测定的需要，仅使用 0.1 mL 样品，显着减少了分析人员的工作时间，纳入了 10 种额外的分析物，并允许更全面的测试制度，以便更好地确定死因。
Systematic toxicological approaches that employ both ideology changes and improvements in instrumentation and sample extraction allow for improved toxicology testing efficiency through lower sensitivities, higher specificity and minimized resource use. Historically, the San Francisco Office of the Chief Medical Examiner relied heavily on a gas chromatography-mass spectrometry (GC-MS) testing regime, comprised of individual drug-class confirmation and quantitation assays. Traditional methods utilizing GC-MS typically require iterations of testing, exhausting sample volume, and hindering productivity and turnaround times, particularly for polypharmacy cases frequently seen in modern postmortem toxicology. The method described here consolidated the scope of seven legacy methods into a single liquid chromatography-tandem mass spectrometry (LC-MS-MS) method for better sensitivity, higher throughput, minimal sample consumption for the quantitation of drugs of abuse and improved quality assurance with the incorporation of smart, automated processing. About 100 mu L of blood or urine were rapidly extracted using a simple acetonitrile protein crash and subsequent in-vial filtration and injected on to an LC-MS-MS system. The developed method was fully validated to SWGTOX and international guidelines and incorporated 55 analytes along with a customized query that facilitates rapid and consistent application of acceptability criteria for data processing and review. Applicability was demonstrated with the analysis of 1,389 samples (858 blood and 531 urine) where at least 41% of positive results may have been missed due to their decreased sensitivity and 11% of results were not within the scope of the previous analytical methods estimated. On average, cases in this study would have previously required three distinct GC-MS assays, 3mL of blood, and upwards of 30 h of active staff time. The described LC-MS-MS analytical approach has mitigated the need to perform multiple assays, utilized only 0.1 mL of sample, significantly reduced analyst work time, incorporated 10 additional analytes and allowed for a more comprehensive testing regime to better inform cause of death determinations.