在线固相萃取-液质联用法同时检测血液样品中12种芬太尼类药物  

Integral SPE-LC-MS/MS to Simultaneously Determine 12 Fentanylkind Drugs in Blood

作　　者：孙立敏[1] 王松才[1] 朱焕慧[1] 林贤文[1] 管旭 谭莉 SUN Limin;WANG Songcai;ZHU Huanhui;LIN Xianwen;GUAN Xu;TAN Li(Guangzhou Institute of Forensic Science,Guangzhou 510030,China)

机构地区：[1]广州市刑事科学技术研究所,广州510030

出　　处：《刑事技术》2022年第2期121-127,共7页Forensic Science and Technology

基　　金：广州市民生科技攻关重大专项(2019030009)。

摘　　要：目的 采用在线固相萃取-液质联用(in-connection SPE-LC-MS/MS)技术,建立血液样品中12种芬太尼类药物的同时检测方法,为刑事案件血液中12种芬太尼类药物的检验鉴定提供依据。方法 血液样品用乙腈沉淀蛋白,经离心、稀释、过滤后上样,采用Agilent PLRP-S(15~20μm,2.1mm×12.5mm)在线SPE柱富集纯化,以Agilent Poroshell 120 EC-C18(3.0mm×150mm,2.7μm)为分析柱,用5mmol/L醋酸铵缓冲液(含0.1%甲酸)-乙腈为流动相进行梯度洗脱,采用电喷雾离子源(ESI)、多反应监测(MRM)对12种芬太尼类药物进行检测。结果 实验优化了色谱条件、质谱条件、在线SPE上样流速、流动相种类、六通阀切换时间、样品有机溶剂比例等参数,经验证,在实验浓度范围内,12种芬太尼类药物线性关系良好,R^(2)均大于0.9990,方法检出限为0.2~0.4ng/mL,定量限为0.7~1.4ng/mL,回收率为83.1%~131.1%,日内精密度和日间精密度分别为1.0%~25.6%、2.1%~30.5%(n=6)。结论 本实验建立的血液样品中的12种芬太尼类药物同时检测方法,具有操作简单方便、灵敏度高、样品需求量少、检出限低等特点,适合于血液样品中芬太尼类药物的检测分析。Objective To establish a method for simultaneous determination of 12 fentanyl-kind drugs in blood through integral SPE-LC-MS/MS with purpose to provide reference for identifi cation of relevant cases. Methods The blood samples(harboring the 12 fentanyl-kind drugs) were treated with acetonitrile for proteins to precipitate, successively centrifugated to collect the supernatant that was subjected to dilution and fi ltration. The fi ltrate was directly undergone into integral SPE-LCMS/MS processing so that Agilent PLRP-S(15-20μm, 2.1mm×12.5mm) column had played its role to enrich and purify the injected fi ltrate and Agilent Poroshell 120 EC-C18(3.0mm×150mm, 2.7μm) column to perform its chromatographic separation under the gradient elution of mobile phases of A(5mmol/L ammonium acetate plus 0.1% formic acid) and B(acetonitrile).The analytes were detected in multiple reaction monitoring(MRM) mode through the electrospray positive ionization(ESI+).Results Under the optimization of chromatographic condition, mass spectral setting, sampling flow, mobile phase, valve swithching programs and organic solvent ratio, the 12 fentanyl-kind drugs showed good linear relationship within the tested concentration ranges, revealing their coeffi cients of determination as R^(2)>0.9990. The method was present of detection limits and quantifi cation limits being 0.2-0.4ng/mL and 0.7-1.4ng/mL, demonstrating the recoveries at three levels being fallen into 83.1%-131.1%, intra-and inter-day RSDs being the respective 1.0%-25.6% and 2.1%-30.5%(n=6). Conclusions The method established here is simple and effi cient, demonstrating its excellence of low quantity of required sample, high sensitivity and low detection limit, suitable for simultaneous determination of 12 fentanyl-kind drugs in blood.

关 键 词：毒物分析 在线固相萃取 液质联用 芬太尼 

分 类 号：DF795.1[医药卫生—法医学]