LC-MS/MS法测定大鼠肝微粒体孵育液中靛玉红的含量

Determination of Indirubin in Rat Hepatomicrosome Incubation Solution Using LC-MS/MS

目的:建立测定大鼠肝微粒体孵育液中靛玉红含量的方法。方法:采用液相色谱-电喷雾串联质谱法。肝微粒体孵育液样品处理采用液-液提取,提取物以甲醇-10mmol·L-1乙酸铵溶液(氨水调pH值至7.40)=90:10为流动相进行分离,以选择性反应离子监测(SRM)方式进行定量分析,用于监测的离子为m/z262.9→219.0(靛玉红)和m/z531.0→82.1(酮康唑,内标)。结果:5min内完成靛玉红的检测,工作曲线线性范围为1～100ng·mL-1,日内、日间精密度分别<3.6%、<13.4%,平均回收率为94.0%～104.4%,检测限为0.5ng·mL-1。结论:本方法灵敏度高、特异性好,可以用于肝微粒体孵育液样品中靛玉红的检测。

OBJECTIVE： To establish LC-MS/MS method for the content determination of indirubin in rat hepatomicrosome in- cubation solution. METHODS ： A liquid-liquid extraction procedure was used for the processing of hepatomicrosome incubation solu- tion through the LC-MS/MS method The extracts were separated by HPLC using methanol-10 mmol-L-~ ammonium acetate （pH val- ue adjusted to 7.40） （90：10） as mobile phase. Selected reaction monitoring （SRM） mode with the transition of m/z 262.9→219.0 （indirubin） and m/z 531.0→82.1 （ketoconazole, IS） was used for quantitative analysis. RESULTS ： The determination of indirubin was completed within 5 min. The linear range of indirubin was 1-100 ng.mL-1 with average recovery of 94.0%-104.4%. The RSD of intra-day and inter-day were lower than 3.6% and 13.4%, and the limit of detection was 0.5 ng-mL -1. CONCLUSION： The method is accurate, sensitive and rapid for the determination of indirubin in rat hepatomicrosome incubation solution.