固相萃取-高效液相色谱法对血液中唑吡坦及其两种羧酸代谢物的同时测定

建立了固相萃取-高效液相色谱分析血液中唑吡坦、6-羧酸唑吡坦及苯基-4-羧酸唑吡坦的方法。采用Waters Oasis HLB 3 mL固相萃取小柱萃取,以乙腈-2.5 mmol/L醋酸铵水溶液为流动相,梯度洗脱,经ZORBAX Eclipse XDB-C18色谱柱(4.6 mm×250 mm,5μm)分离,二极管阵列检测器(DAD)检测,检测波长为250 nm,外标法定量。比较了不同洗脱溶剂、溶液pH值对回收率的影响。在优化条件下,唑吡坦、6-羧酸唑吡坦及苯基-4-羧酸唑吡坦的线性范围分别为40~2000μg/L(r=0.9999)、20~2000μg/L(r=0.9998)和40~2 000μg/L(r=0.9998),检出限为1.2~3.3μg/L。添加16、160、360μg/L 3个水平的混合标准溶液,3种化合物的回收率为86%~106%,相对标准偏差均小于8%。结果表明,该方法具有操作简便、回收率高、精密度好和灵敏度高等特点,适于血液中唑吡坦及其2个羧酸代谢物的分析,可为相关案件提供方法及技术支持。

Simultaneous Determination of Zolpidem and Its Two Carboxyl Metabolites in Blood by High Performance Liquid Chromatography with Solid Phase Extraction

A solid phase extraction-high performance liquid chromatographic method was developed for analysis of zolpidem,zolpidem 6-carboxylic acid(ZCA) and zolpidem phenyl-4-carboxylic acid(ZPCA) in blood sample.The sample was extracted by Waters Oasis~(R) HLB 3 mL solid phase column and separated on an ZORBAX Eclipse XDB-C_(18) column(4.6 mm×250 mm,5 μm) under gradient condition using acetonitrile and 2.5 mmol/L ammonium acetate water solution as the mobile phase.Analysis of zolpidem,ZCA and ZPCA were performed by photo diode array detection(DAD) at 250 nm with external standard method.The eluting solvent and pH value of solution on the recoveries were investigated and the chromatogramic conditions were optimized.Under the optimal condition,the method showed good linearity in the ranges of 40-2 000 μg/L for zolpidem(r=0.999 9),20-2 000 μg/L for ZCA(r=0.999 8),and 40-2 000 μg/L for ZPCA(r=0.999 8).The limits of detection were between 1.2 and 3.3 μg/L.The recoveries of zolpidem and two carboxyl metabolites were from 86% to 106% at three spiked levels of 16,160 and 360 μg/L.The relative standard de- viations(RSDs) of the method were less than 8%.The results indicated that this method was simple,accurate and sensitive,and was suitable for the analysis of zolpidem and its two carboxyl metabolites in the blood sample.