液相色谱-串联质谱法测定人血浆中的龙胆苦苷浓度

Determination of gentiopicroside in human plasma by liquid chromatography-tandem mass spectrometry

目的:建立测定人血浆中龙胆苦苷浓度的液相色谱-串联质谱法。方法:血浆加入内标咖啡因后经固相萃取处理,采用 RESCEK C_8柱(150 mm×2.1 mm,5μm)分离,流动相为甲醇-10 mmol·L^(-1)醋酸铵溶液-乙腈(50:40:10),流速为0.2mL·min^(-1)。样品在三级四极杆串联质谱中经 ESI 源离子化后以多反应离子监测方式测定。结果:龙胆苦苷在3～5000 ng·mL^(-1)线性良好(r=0.9985),检测限为3 ng·mL^(-1),回收率为94.4%～104.2%,绝对回收率为92.4%～98.0%,日内、日间变异(RSD)均≤15%,色谱峰保留时间为2.25 min。结论:方法灵敏、准确、快速、特异性强,适用于中药龙胆苦苷的血药浓度测定和临床药代动力学研究。

Objective：To establish an HPLC/MS/MS method in order to determine gentiopicroside in human plasma. Method：The plasma sample by solid -phase extraction with caffeine as internal standard, used the RESCEK Cs column（ 150 mm × 2. 1 mm,5 μm）as analytical column, the mobile phase consisted of methanol- 10 mmol · L^-1 ammonium acetate buffer - acetonitrile （50： 40： 10）with the flow rate of 0. 2 mL · min^-1. The sample was ionized by electrospray ionization（ESI） source in the triple quadruple tandem mass spectrometer, and was quantitated with multiple reaction monitoring mode. Results： The good linear range of gentiopicroside was 3 - 5000 ng · mL^-1 （ r = 0. 9985 ） ,the limit of quantification was 3 ng · mL^-1 ,the recovery and the absolute recovery was 94. 4% - 104.2% and 92. 4% -98.0% ,respectively. The within- day and between- day RSD were less than 15% ,the chromatography peak tR was 2. 25 min. Conclusion：The method is sensitive, accurate, rapid,, specific, suitable for determination of gentiopicroside in human plasma and clinical pharmacokinetic study.