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目的:建立血浆中甘草苷、甘草素、异甘草苷、异甘草素、甘草酸和甘草次酸的LC-MS/MS分析测定法,研究半夏泻心汤[由半夏和干姜(辛开组),黄连和黄芩(苦降组),人参、大枣和炙甘草(甘补组)组成]及不同配伍组中甘草活性成分在大鼠体内的药代动力学。方法:取血浆样品0.1 mL经甲醇-丙酮-乙酸乙酯(5∶4∶1)沉淀蛋白并萃取后,经Inertsil ODS-SP色谱柱(100 mm×2.1 mm,5μm)分离,流动相为甲醇-0.1%甲酸(含5 mmol.L-1醋酸铵),梯度洗脱。采用电喷雾电离源,多反应监测模式(MRM)测定各成分的血药浓度,DAS 2.0计算药动学参数,SPSS 17.0统计分析。结果:血浆中6个甘草活性成分的提取回收率均大于78%;LC-MS/MS方法测定的日内和日间精密度RSD均小于12%;药动学结果显示,全方配伍后,甘草苷和甘草素的AUC(0-∞)显著高于甘补组和甘补苦降组,相应CL/F显著性降低;异甘草素的Cmax和AUC(0-∞)显著性提高;甘草次酸的Cmax和AUC(0-∞)均高于甘补辛开组和甘补苦降组,Cmax分别高达1.93和4.08倍,AUC(0-∞)分别高达2.49和4.80倍。而甘草酸的AUC(0-∞)在甘补苦降配伍中较高,甘草次酸的AUC(0-∞)则在甘补组中较高。结论:建立的LC-MS/MS分析方法灵敏、准确,可用于半夏泻心汤中活性成分的药代动力学研究。结果表明大鼠口服半夏泻心汤及不同配伍后,各活性成分的药动学参数有一定的变化,全方配伍利于多数活性成分的吸收。
Objective: To establish a LC-MS / MS method for the determination of glycyrrhizin, glycyrrhizin, isoliquiritin, isoliquiritigenin, glycyrrhizin and glycyrrhetinic acid in plasma, Group), Coptis and Scutellaria (bitter group), ginseng, jujube and Zhigancao (Ganbu group)] and different compatibility group licorice root pharmacokinetics in rats. Methods: The plasma samples were extracted with 0.1 mL methanol-acetone-ethyl acetate (5: 4: 1) and extracted by Inertsil ODS-SP column (100 mm × 2.1 mm, 5 μm). The mobile phase consisted of methanol -0.1% formic acid (containing 5 mmol.L-1 ammonium acetate), gradient elution. The plasma concentration of each component was determined by electrospray ionization source and multi-reaction monitoring mode (MRM). The pharmacokinetic parameters were calculated by DAS 2.0 and SPSS 17.0 statistical analysis. Results: The recoveries of 6 licorice active ingredients in plasma were both higher than 78%. The RSDs of intra-and inter-day precision were both less than 12% by LC-MS / MS. The pharmacokinetic results showed that after the whole prescription was combined, The AUC (0-∞) of glycosides and glycyrrhizin were significantly higher than those of Ganfu group and Ganbu bitterness group, and the corresponding CL / F was significantly decreased; the Cmax and AUC (0-∞) of isoliquiritigenin were significantly increased; The Cmax and AUC (0-∞) of acid were higher than that of Ganduxin group and Gandu biting group, respectively. The Cmax values were as high as 1.93 and 4.08 times, respectively. The AUC (0-∞) were 2.49 and 4.80 times respectively. However, the AUC (0-∞) of glycyrrhizic acid was higher in the combination of Gan Bu Bu Kuangding, and the AUC (0-∞) of glycyrrhetinic acid was higher in Ganbu group. Conclusion: The established LC-MS / MS method is sensitive and accurate and can be used to study the pharmacokinetics of the active ingredients in Banxiaxiexin Decoction. The results showed that rats after oral administration of Banxiaxiexin Decoction and different compatibility, the pharmacokinetic parameters of each active ingredient has a certain degree of change, the whole compatibility is conducive to the absorption of most active ingredients.