UPLC-MS/MS Method for Detection of Etomidate and Its Metabolite Etomidate Acid Quantity in Blood.

Journal of Forensic Medicine Pub Date : 2023-12-25 DOI:10.12116/j.issn.1004-5619.2023.330901

Xing Han, Xin Liu, Ming-Luo DU, Ruo-Lun Xu, Jia-Rong Li, Chao Liu, Wei-Guo Liu

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Abstract

Objectives: To establish a method for the simultaneous quantitative analysis of etomidate and its metabolite etomidate acid in blood, and to discuss its application value in actual cases.

Methods: Acetonitrile precipitate protein method was used, and C₁₈ column was selected. Gradient elution was performed with acetonitrile and 5 mmol/L ammonium acetate within 6 min. Electrospray ionization source in positive ion mode was used. The internal standard etomidate acid-d₅ was obtained by etomidate-d₅ alkaline hydrolysis reaction. Ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was used for quantitative analysis. The methodological verification was conducted.

Results: Etomidate and etomidate acid in blood showed good linear relationship in the quantitative linear range (r>0.999), with the lower limit of quantification was 2.5 ng/mL and 7.5 ng/mL, respectively. The accuracy, precision, recovery rate, and matrix effect of the method met the professional verification standards. The practical application results showed that etomidate and etomidate acid could be detected in the blood of the abusers, and their mass concentrations ranged from 17.24 to 379.93 ng/mL.

Conclusions: The method established in this study can simultaneously quantify etomidate and etomidate acid in blood, which is simple and convenient to operate with accuracy. It can meet the detection needs of actual cases and provide technical support for law enforcement to crack down on etomidate abuse.

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用于检测血液中依托咪酯及其代谢物依托咪酯酸含量的 UPLC-MS/MS 方法。

目的建立血液中依托咪酯及其代谢产物依托咪酯酸的同时定量分析方法，并探讨其在实际案例中的应用价值：方法：采用乙腈沉淀蛋白法，C18色谱柱。采用乙腈沉淀蛋白法，选用C18色谱柱，以乙腈和5 mmol/L乙酸铵在6 min内梯度洗脱。电喷雾离子源为正离子模式。内标依托咪酯酸-d5 由依托咪酯-d5 碱水解反应得到。采用超高效液相色谱-串联质谱（UPLC-MS/MS）进行定量分析。进行了方法学验证：结果：血液中依托咪酯和依托咪酯酸在定量线性范围内呈良好的线性关系（r>0.999），定量下限分别为2.5 ng/mL和7.5 ng/mL。该方法的准确度、精密度、回收率和基质效应均符合专业验证标准。实际应用结果表明，在滥用者血液中可检测到依托咪酯和依托咪酯酸，其质量浓度范围为17.24～379.93 ng/mL：本研究建立的方法可同时定量检测血液中的依托咪酯和依托咪酯酸，操作简单方便，结果准确。可满足实际案件的检测需要，为执法部门打击依托咪酯滥用行为提供技术支持。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Journal of Forensic Medicine

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