UPLC-MS/MS法研究小檗碱、巴马汀和药根碱在大鼠体内的组织分布

Tissue distribution of berberine,palmatine and jateorhizine in rat determined by UPLC-MS/MS

目的:建立UPLC-MS/MS同时测定大鼠组织中小檗碱、巴马汀和药根碱3个生物碱成分的分析方法,并应用于这3个成分在大鼠体内的组织分布研究。方法:采用BEH C18色谱柱(2.1 mm×50 mm,1.7μm),以乙腈-0.1%甲酸水溶液(20∶80)为流动相,流速0.3 m L·min^(-1),正离子多离子反应监测(MRM)检测。组织样品匀浆后乙腈沉淀蛋白供测定。结果:在线性范围内3个成分的线性关系良好,准确度、日内和日间精密度、稳定性、提取回收率均符合生物样品的分析要求。3个生物碱成分在大鼠体内分布广泛,胃、小肠和肝组织中浓度较高,在心脏中的分布浓度最低。结论:本文方法可用于黄连提取物在大鼠体内的组织分布研究。

Objective： To develop a UPLC-MS/MS method for simultaneous determination of berberine, palmatine and jateorhizine in rat tissues, and apply the method to tissue distribution study of three alkaloids. Methods： The UPLC gradient elution was performed using BEH C18 （ 2.1 mm ×50 mm, 1.7 μ m ） column with the mobile phase of acetonitrile-0.1% formic acid water solution （ 20 ： 80 ） at the flow rate of 0.3 mL min-1. The mass spectrometer was operated in Multi ion reaction monitoring （ MRM ） mode with positive electrospray ionization. The tissue samples were homogenated and the suspension was extracted by precipitating with aeetonitrile for determination. Results： The relationships between the concentration and the peak areas of the three compounds were all linear. The preeisions, accuracy, stability, extract recoveries and stability of the analytes met the biological requirements. Berberine, palmatine and jateorhizine were widely distributed in rat tissues. High concentrations were found in stomach, small intestine and liver, and the distribution concentration was the lowest in heart. Conclusion： The method can be used to study the tissue distribution of Rhizoma Coptidis extract in rats.