人毛发中36种芬太尼类物质的HPLC-MS/MS检测方法

Determination of 36 Fentanyl Analogs in Human Hair by HPLC-MS/MS

建立了同时对人毛发中36种芬太尼类物质快速定性定量检测方法,并成功应用于实际案件的检测。利用高效液相色谱-串联质谱(HPLC-MS/MS)技术,采用MRM(多反应监测)模式,用Waters ACQUITY BEH C18(100 mm×2.1 mm,1.7μm)色谱柱,柱温50℃,流动相为甲醇-0.1%甲酸的水溶液(V/V),梯度洗脱。毛发样品采用0.1%SDS、水、丙酮依次清洗后,研磨成粉末,称取约20 mg,加1 mL甲醇浸泡18 h,后再加入1 mL去离子水,混匀后过滤膜,用液相色谱-串联质谱进行检测。36种芬太尼类物质的检出限均为0.01 ng·mg^-1,保留时间RSD均小于0.33%。对芬太尼和阿芬太尼进行了定量分析,在0.01~2 ng·mg^-1范围内均具有良好的线性关系,判定系数R2分别为0.997和1,在0.05、0.2和1 ng·mg^-1的添加水平的回收率为89.34%~104.80%,基质效应为96.95%~112.59%,准确度范围为91.92%~102.34%,日内精密度范围为0.49%~6.84%,日间精密度范围为1.21%~7.00%。该方法简单、高效、准确、灵敏,可作为芬太尼类物质滥用情况的监测方法。

Objective A rapid qualitative and quantitative method for the detection of 36 fentanyl analogs in human hair simultaneously was established,and this method was applied in actual cases.Methods The hair sample was determined by HPLC-MS/MS using MRM(multiple reaction monitor)mode,and separated on chromatographic column Waters ACQUITY BEH C18(100 mm×2.1 mm,1.7μm)with mobile phase consisting of methanol-aqueous solution of 0.1%formic acid(V/V),gradient elution.The hair samples were washed with 0.1%SDS,water and acetone successively.Then the hair samples were ground into a powder,weighted 20 mg,soaked with 1mL methanol for 18h,added 1mL deionized water,mixed and filtrated the solution with filter membrane,analyzed the solution using HPLC-MS/MS.Results 36 fentanyl analogs can be separated effectively,including that baseline separation of 3 pairs of isomers were achieved.The detection limits of 36 fentanyl analogs were all 0.01 ng·mg^-1.The retention time RSDs were less than 0.33%.The quantitative detection was conducted on Fentanyl and Alfentanil,the linear ranges are all 0.01~2 ng.mg^-1,and the correlation coefficient were 0.997 and 1 respectively.When the spiked levels were 0.05、0.2、1 ng·mg^-1,the average recoveries were between 89.34%~104.80%,the matrix effect was in the range of 96.95%~112.59%,the rang of accuracy was 91.92%~102.34%,the ranges of intraday and interday relative standard deviations(RSDs)were 0.49%~6.84%and 1.21%~7.00%respectively.Conclusion The method is simple,efficient,accurate and sensitive and it can be used as a monitoring method for the abuse of fentanyl substance.