手性柱液质联用法同时测定人体尿液中 泮托拉唑对映体浓度*

目的:建立同时测定人体尿液中左旋泮托拉唑和右旋泮托拉唑浓度的手性柱-液质联用法(Chiral-LC-MS)。方法:以乙腈作为蛋白质沉淀剂,对尿液中的泮托拉唑对映异构体进行Chiral-LC-MS分析。色谱柱:Chiral PAK IE(4.6 mm×150 mm,5μm),柱温为30℃,流动相为乙腈-10 mmol·L^-1乙酸铵(含0.1%乙酸)(72∶28,v/v),泮托拉唑对映异构体在9.0 min内可达到基线分离。使用电喷雾离子化(ESI)正离子模式,泮托拉唑和内标((R)-Pantoprazole-d6)的多反应监测(MRM)扫描离子对分别为m/z 384.1→200.1和390.1→206.0。结果:该方法无明显的基质效应,左旋泮托拉唑和右旋泮托拉唑在标准曲线范围内线性关系良好,均为2.50~5000 ng·mL^-1,批内、批间准确度和精密度均<15%。在泮托拉唑对映体的储存、处理和分析过程中未发现两者间的手性转化,测定过程中各化合物稳定性符合要求。结论:建立了简单、高选择性的Chiral-LC-MS方法,可用于同时测定人体尿液中的泮托拉唑对映体的浓度。

Determination of Pantoprazole Enantiomers in Human Urine by a Chiral-LC-MS Method*

Objective:A simple enantioselective method was developed and validated for simultaneous determination of S-and R-pantoprazole in human urine by chiral liquid chromatography with tandem mass spectrometry(Chiral-LC-MS).Methods:Pantoprazole enantiomers and internal standard((R)-pantoprazole-d6)were extracted from urine using acetonitrile as the protein precipitating agent.Baseline chiral separation was achieved within 9.0 min on a Chiral PAK IE column(4.6 mm×150 mm,5μm)at 30℃.The mobile phase was consisted of acetonitrile-10 mmol·L^-1 ammonium acetate solution containing 0.1%acetic acid(72∶28,v/v).Using the electrospray ionization(ESI)positive ion mode,multiple reaction monitoring(MRM)scanning ion pairs for pantoprazole and(R)-pantoprazole-d6 were m/z 384.1→200.1 and 390.1→206,respectively.Results:For each enantiomer,no apparent matrix effect was found.All the analytes showed good linearity between 2.50-5000 ng·mL^-1.The RSD for intra-and inter-precisions and accuracy were within 15.0%.No chiral inversion was observed during the procedures of sample storage,preparation or analysis.Analytes were stable during the study.Conclusion:A simple and selective chiral-LC-MS method has been successfully developed for the simultaneous determination of pantoprazole enantiomers in human urine.