LC/MS/MS测定龙胆苦苷的尿药浓度

Determination of Gentiopicroside in Urine by LC/MS/MS

目的建立龙胆苦苷尿药浓度的液质联用测定方法(LCM/SM/S)。方法以咖啡因作内标,固相萃取法处理尿样。色谱柱:RESCEK C8柱(150mm×2.1mm,5μm),流动相为甲醇-10mm ol.L-1醋酸铵缓冲液(pH=6.5)-乙腈(50∶40∶10),流速为0.2m L.min-1。样品经电喷雾离子源(ESI)正离子化后,通过三级四极杆串联质谱仪,采用多反应离子监测方式测定龙胆苦苷(mz/374.1→195.2)和咖啡因(m z/195.2→138.2)浓度。结果尿液中龙胆苦苷在30～9000ng.mL-1浓度范围内线性关系良好r(=0.9980),方法回收率为91.10%～96.21%,提取回收率为100.52%～103.83%,日内、日间精密度均<10%。结论该方法灵敏、准确、快速、特异性强。

Objective To establish a HPLC/MS/MS method for the determination of gentiopicroside in human urine. Methods The urine sample was treated by solid-phase extraction with internal standard of caffeine; The RESCEK C8 column （150 mm×2.1 mm, 5μm） was used as the analytical column with a mobile phase consisting of methanol-10mmol·L^-1NH4AC buffer （pH=6.5）-acetonitrile （50：40：10, V/V） , the flow rate was 0.2mL·min^-1; A triple quadruple tandem mass spectrometer was used as the detector, Electrospray ionization source was applied and operated in positive ion mode. Gentiopicroside and caffeine were detected by monitoring the ion transition of m/z 374.1→195.2 and m/z 195.2→138.2 respectively. Results The linear range was 30-9000 ng·mL^-1（r=0.9980） for gentiopicroside in human urine. The recovery was 91.10%-96.21%, The absolute recovery was 100.52%-103.83%, The within-day and between-day precisions were less than 10%. Conclusion The method is proved to be sensitive, accurate, rapid, specific.