LC-MS/MS同时测定大鼠血浆中4种黄连生物碱的含量

Simultaneous Determination of Four Kinds of Alkaloid from Rhizoma Coptidis in Rat Plasma by LC-MS/MS

目的:建立LC-MS/MS同时测定大鼠血浆中黄连碱、药根碱、巴马汀和小檗碱含量的方法。方法:采用ACQUITYUPLC BEH C18色谱柱(2.1 mm×50 mm,1.7μm),流动相乙腈-水(含2 mmoL乙酸铵和0.05%甲酸),流速0.3 mL.min-1,柱温30℃,以士的宁为内标,血浆样品用4倍量乙腈萃取并沉淀蛋白,用电喷雾离子化(ESI)和正离子多离子反应监测(MRM)模式检测,选择离子对为药根碱m/z 338.227→m/z 322.824;黄连碱m/z 320.244→m/z 291.996;巴马汀:m/z 352.243→m/z308.016;小檗碱:m/z 336.227→m/z 277.981;士的宁:m/z 335.264→m/z 184.017。结果:药根碱、黄连碱、巴马汀和小檗碱的线性范围分别为1.01～505,1.06～530,1.13～565,1.07～535μg.L-1;定量下限分别为0.539 2,0.646 3,0.761 5,0.197 1μg.L-1;平均回收率分别为103.90%,101.50%,101.58%,115.54%;日内和日间精密度、稳定性的RSD均<10%。结论:建立的方法准确、灵敏、特异、简便,适用于大鼠血浆中黄连碱、药根碱、巴马汀和小檗碱的同时测定及药动学研究。

Objective： To develop a LC-MS/MS method for simultaneous determination of jatrorrhizine,coptisine,palmatine and berberine in rat plasma.Method： Taking ACQUITY UPLC BEH C18column（2.1 mm × 50 mm,1.7 μm） with mobile phase of acetonitrile-water（2 mmoL ammonium acetate and 0.05% formic acid）,flow rate was 0.3 mL.min-1,column temperature was 30 ℃.Strychnine was adopted as the internal standard,plasma samples were pretreated by protein precipitation with 4 times the amount of acetonitrile,electrospray ionization（ESI） was applied and operated in positive ionmode,quantification was performed by multiple reaction monitoring（MRM） of jatrorrhizine m / z 338.227→m/z 322.824,coptisine m/z 320.244→m/z 291.996,palmatine m / z 352.243→ m/z 308.016,berberine m/z 336.227→ m/z 277.981 and the internal standard m / z 335.264 → m / z 184.017,respectively.Result： Calibration curve of jatrorrhizine,coptisine,palmatine and berberine in plasma were linear in the range of 1.01-505,1.06-530,1.13-565,1.07-535 μg.L-1;Lower limit of quantitation were 0.539 2,0.646 3,0.761 5 and 0.197 1 μg.L-1,respectively;Average recoveries were in order of 103.90%,101.50%,101.58% and 115.54%,respectively.RSD of intra-and inter-day,stability were less than 10%.Conclusion： This assay method was accurate,sensitive,specific and suitable for simultaneous determination and pharmacokinetic study of jatrorrhizine,coptisine,palmatine and berberine in rat plasma.