基于UPLC-MS/MS大承气汤多种活性成分大鼠体内药动学研究

In vivo pharmacokinetic study on several active components of Dachengqi Decoction in rats by UPLC-MS/MS

目的建立超高效液相色谱-串联质谱法（UPLC-MS/MS）,研究大鼠ig大承气汤后芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚、柚皮苷、橙皮苷、新橙皮苷、和厚朴酚及厚朴酚10种有效成分的体内药动学过程。方法大鼠ig大承气汤（10.35 g/kg）后,于不同时间点从眼底静脉丛取血,以1,8-二羟基蒽醌为内标,血浆样品前处理后进行UPLC-MS/MS分析。以甲醇-0.1%甲酸为流动相梯度洗脱,色谱柱为岛津Shim-pack XR-ODS III（75 mm×2.0mm,1.6μm）,体积流量0.35mL/min,采用电喷雾离子化（ESI）源,负离子模式扫描,多反应监测模式（MRM）检测各有效成分;采用Win Nonlin 4.1药动学软件计算药动学参数。结果芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚、柚皮苷、橙皮苷、新橙皮苷、和厚朴酚、厚朴酚10种有效成分质量浓度分别在0.52-520.00、2.06-616.00、0.57-568.00、3.1-1 240.0、18.5-1 000.0、1.94-972.00、1.59-796.00、1.63-816.00、0.025-50.000、0.027-53.300 ng/mL线性关系良好。各成分精密度、回收率良好,且在整个分析过程中稳定,均符合生物样品分析要求。给药大鼠血浆中大黄素甲醚大多数点血药浓度低于检测限,未能得到药动学参数;其他成分均得到完整的血药浓度-时间曲线和药动学参数。结论该方法可用于同时测定大承气汤中多种有效成分的血药浓度,可用于药动学研究,且方法准确。

Objective To study the in vivopharmacokinetics of aloe emodin, rhein, emodin, chrysophanol, physcion, hesperidin, neohesperidin, naringin, honokiol, and magnolol in plasma of rats after ig administration of Dachengqi Decoction. Methods Afterig administration of Dachengqi Decoction（10.35 g/kg）, the blood was collected from theretinal vein plexus of rats at predetermined time and pre-prepared with 1,8-dihydroxyanthraquinone as internal standard（IS）. A reliable ESI source, negative ion mode scanning, and multiple reaction monitoring（MRM） method were developed for thesimultaneous determination of the active components in Dachengqi Decoction and chromatography was carried out on a Shimadzu Shim-pack XR-ODS III（75mm ×2.0 mm, 1.6μm） using a gradient mobile phase consisted of methanol and 0.1% formic acid water at a flow rate of 0.35 mL /min. Calculations of the pharmacokinetic data were performed using Win Nonlin 4.1software. Results Calibration curves for aloe emodin, rhein, emodin, chrysophanol, physcion, hesperidin, neohesperidin, naringin, honokiol, and magnolol in plasma of rats were linear within the ranges of 0.52—520.00, 2.06—616.00, 0.57—568.00, 3.1—1240.0, 18.5—1000.0, 1.94—972.00, 1.59—796.00, 1.63—816.00, 0.025—50.000, and 0.027—53.300 ng/mL, respectively. The precision, recovery, and stability of each component in the present assay were acceptable for the analysis. They are in the line with the requirements of biological sample analysis. The concentration of physcion in plasma of rats was under the limit of detection at almost every time point, thus we failed to obtain the pharmacokinetic parameters of it. While the plasma concentration and pharmacokinetic parameters of other components were successfully calculated. Conclusion The proposed method can be used forthesimultaneous determination of the concentration of several active components in plasma of rats and successfully applied to the pharmacokinetic study of the analytes in rats afterig administration of Dachengqi Decoction.