UPLC-MS/MS法测定血液中依托咪酯及其代谢物依托咪酯酸的含量

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

目的 建立血液中依托咪酯及其代谢物依托咪酯酸的同步定量分析方法 ,探讨其在实际案件中的应用价值。方法 采用乙腈沉淀蛋白法,选用C_(18)色谱柱,以乙腈和5 mmol/L乙酸铵水溶液在6 min内进行梯度洗脱,采用电喷雾离子源、正离子模式,通过依托咪酯-d_5碱水解反应获得内标依托咪酯酸-d_5,使用超高效液相色谱-串联质谱法(UPLC-MS/MS)定量,并进行方法 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线性关系良好(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、基质效应等参数验证结果 均满足行业验证标准。实际应用结果 表明,依托咪酯滥用者血液中能够检出依托咪酯和依托咪酯酸两种成分,其质量浓度范围为17.24~379.93 ng/mL。结论 本方法可同步定量血液中的依托咪酯及依托咪酯酸,操作简便、定量准确,能够满足实际案件的检测需求,可为执法机关打击依托咪酯滥用行为提供技术支持。

Objective 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.Re‐sults 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.Conclu‐sion 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.