LC-MS-MS测定黄连解毒汤中3种生物碱在大鼠血清的含量及其药代动力学研究

Pharmacokinetics of three alkaloids in Huanglianjiedu dcoction in rat serum by LC-MS-MS

目的：建立灵敏、特异、准确、可靠的药根碱、巴马汀、小檗碱在大鼠血清中液质联用（LC-MS-MS）分析方法，用于黄连解毒汤中3种生物碱在正常大鼠体内药代动力学研究。方法：优化药根碱、巴马汀、小檗碱及内标延胡索乙素质谱检测条件，确定相关条件并开展方法学考察。正常大鼠灌胃给予黄连解毒汤，经时取血，样品经处理后将该方法用于3种成分的测定。DAS药代动力学软件拟合房室模型，计算药动参数。结果：在ESI（＋）离子化条件下采用MRM工作模式，用m／z 338～322，351．9～308，336—319．8来分别检测药根碱、巴马汀、小檗碱，同时以m／z 356．1—192．1来检测内标延胡索乙素。药根碱在0．2—25μg·L^-1、巴马汀、小檗碱在0．4～50μg·L^-1呈良好线性关系。3种生物碱在高、中、低3个浓度的准确度、精密度、稳定性等均符合要求。药根碱、巴马汀、小檗碱在正常大鼠体内药动学过程符合一室开放模型。结论：应用LC—MS—S技术建立了测定药根碱、巴马汀、小檗碱血药浓度的方法，并成功应用于黄连解毒汤正常大鼠体内这3个成分的药动学研究，为中药复方药动学研究提供了可以借鉴的分析方法。

Objective： To develop a sensitive and reliable high-performance liquid chromatography-tandem mass sepctrometry （LC-MS-MS） method for simultaneous determination of jatrorrhizine, palmatine and berberine in rat serum, and study the pharmacokinetics of the three alkaloids in rat serum after an intragastrical （ig） administration of Huanglianjiedu decoction to rat. Method： The optimal ionization and fragmentation conditions, as well as HPLC ones, to detect jatrorrhozine, palmatine and berberine were developed and validated. After the Huanglianjiedu decoction were administered to rats through ig route, LC-MS-MS method has been applied to the pharmacokinetic study of the three alkaloids in rat serum. DAS procedure was used to process concentration-time data. Result： The detection was performed by MRM mode via electrospray ionization （ESI） source operating in the positive ionization mode. The precursor-to-product ion transitions were at m/z 338-322 for jatrorrhozine, m/z 351.9-308 for palmatine, m/z 336-319.8 for berberine and m/z 356. 1-192. 1 for tetrahydropalmatine （IS） respectively. The method was linear over the concentration range of 0. 2-25 μg·L^-1 for jatrorrhozine, 0. 4-50 μg·L^- 1 for palmatine and berberine. The method was validated according to the requirements. The pharmacokinetic process of the three alkaloids after oral administration of Huanglianjiedu decoction was fitted to be a one-compartment model. Conclusion： The fully validated LC-MS-MS method has been successfully applied to the pharmacokinetic study of the three alkaloids in rat serum after oral administration of Huanglianjiedu decoction.