大鼠口服半夏泻心汤及不同配伍组中甘草活性成分的药代动力学研究

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 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,5 μm)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C_max和AUC_(0-∞)显著性提高;甘草次酸的C_max和AUC_(0-∞)均高于甘补辛开组和甘补苦降组,C_max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 rapid LC-MS/MS method for simultaneous determination of liquiritin, liquiritigenin, isoliquiritin, isoliquiritigenin, glycyrrhizic acid and glycyrrhetinic acid in rat plasma,so as to study pharmacokinetic process of the active ingredients of Banxia Xiexin decoction and its compatibility groups. Methods: Six components of Banxia Xiexin decoction and internal standard carbamazepine were extracted from blood samples by protein precipitation and methanol-acetone-ethyl acetate(5: 4: 1) extraction,and then separated on an Inertsil ODS-SP column(100 mm×2.1 mm,5 μm) with gradient elution of methanol-0.1% formic acid (including 5 mmol·L^-1 ammonium acetate).Electrospray ionization(ESI) source was applied under the negative ion mode,and multi-reaction monitoring mode(MRM) was used to qualify the analytes in plasma samples.Then DAS 2.0 and SPSS 17.0 software were selected to calculate and analyze the pharmacokinetic parameters of each analyte. Results: The average recoveries of six active ingredients were more than 78%,and the RSDs of intra-and inter-day precisions of all analytes were less than 12%.The pharmacokinetic results showed that after oral administration of Banxia Xiexin decoction,AUC_(0-∞)of liquiritin and isoliquiritigenin increased significantly compared with sweet and sweet-bitter groups,while CL/F decreased accordingly;C_max and AUC_(0-∞) of isoliquiritigenin increased significantly;meanwhile,compared to sweet-pungent and sweet-bitter groups,C_max of glycyrrhetinic acid was improved by 1.93 and 4.08 folds and AUC_(0-∞) was improved by 2.49 and 4.80 folds respectively.Besides,glycyrrhizic acid in sweet-bitter group and glycyrrhetinic acid in sweet group showed higher AUC_(0-∞)values than other compatibilities. Conclusions: This method is of high sensitivity,accuracy and can be successfully applied to the pharmacokinetic study of active components of Banxia Xiexin decoction.The results indicate that after oral administration of different combinations of Banxia Xiexin decoction,some pharmacokinetics parameters of active ingredients are significantly changed,and the bioavailability of most components are improved with Banxia Xiexin decoction is oral administratration.