Wang Jinglong, Zheng Dandan, Wang Yingzi, Zhang Chao, Sun Xiumei
College of Life Sciences, Zaozhuang University, Zaozhuang 277160, China.
College of Life Sciences, Zaozhuang University, Zaozhuang 277160, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2017 Aug 1;1059:35-42. doi: 10.1016/j.jchromb.2017.05.019. Epub 2017 May 24.
To study the pharmacokinetics of Erhuang decoction extracts, a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method was established for the determination of effective substances in rat plasma. The extracts prepared by water extraction (WE) method were given to rats by oral administration. After collected from the orbital venous plexus, plasma was treated by protein precipitation method. Then, the concentration of index components, including baicalin, liquiritin, berberine, palmatine and glycyrrhetinic acid, were determined by HPLC-MS/MS. Gradient elution mode was used to the chromatographic separation with an Inertsil ODS-SP column (100 mm×2.1mm, 5μm), with acetonitrile and 0.1% formic acid containing 10mmolL ammonium acetate as the mobile phase. MS analysis was conducted by multiple reactions monitoring (MRM) with Electrospray Ionization (ESI). The extraction recoveries of the five active ingredients from plasma were greater than 86.04%, and the intra- and inter-day precisions were less than 16.57%. Results indicated that active ingredients in plasma of rats with oral administration of extracts showed certain difference in the pharmacokinetic parameters, which proved that the active ingredients were effectively absorbed. The established HPLC-MS/MS analytical method was sensitive and accurate, suitable for the pharmacokinetic study of active ingredients in Erhuang decoction.
为研究二黄汤提取物的药代动力学，建立了高效液相色谱 - 串联质谱法（HPLC-MS/MS）用于测定大鼠血浆中的有效成分。采用水提法制备的提取物经口服给予大鼠。从眶静脉丛采血后，血浆采用蛋白沉淀法处理。然后，通过HPLC-MS/MS测定指标成分黄芩苷、甘草苷、小檗碱、巴马汀和甘草次酸的浓度。采用梯度洗脱模式，使用Inertsil ODS-SP柱（100 mm×2.1mm，5μm）进行色谱分离，以乙腈和含10mmol/L醋酸铵的0.1%甲酸为流动相。通过电喷雾电离（ESI）的多反应监测（MRM）进行质谱分析。五种活性成分在血浆中的提取回收率大于86.04%，日内和日间精密度小于16.57%。结果表明，口服提取物的大鼠血浆中活性成分的药代动力学参数存在一定差异，证明活性成分被有效吸收。所建立的HPLC-MS/MS分析方法灵敏、准确，适用于二黄汤活性成分的药代动力学研究。
