在线SPE-HPLC系统在补骨脂二氢黄酮甲醚药代动力学研究中的应用

Application of online SPE-HPLC system in pharmacokinetic study of bavachinin

目的:探讨在线固相萃取(SPE)-高效液相色谱(HPLC)系统在小鼠血浆中补骨脂二氢黄酮甲醚药代动力学研究的应用。方法:基于在线SPE-HPLC技术的Ultimate 3000系统测定补骨脂二氢黄酮甲醚血药浓度。色谱条件:采用Venusil MPC18(150 mm×4.6 mm,5μm)色谱柱,流动相为乙腈-5 mmol·L-1磷酸二氢钾缓冲液(pH 3.5),流速1.0 mL·min-1,梯度洗脱;使用Capcell MF Ph-1在线SPE柱(10 mm×4 mm,5μm),洗脱剂为5 mmol·L-1磷酸二氢钾缓冲液(pH 3.5);检测波长238 nm。WinNonlin 5.2软件计算药动学参数。结果:补骨脂二氢黄酮甲醚线性范围为20.0～4000 ng·mL-1,最低定量限为20.0 ng·mL-1,提取回收率为90.5%～94.6%,日内、日间精密度试验的RSD均小于10%,短期稳定性、冻融稳定性及长期稳定性准确度为92.38%～101.7%。结论:该方法工作流程简化,样品分析快速,提高了工作效能,其结果可信且重复性好。

Objective：To study the application of online solid phase extraction - high performance liquid chromatography （ SPE - HPLC ） systems in the plasma pharmacokinetic study of bavaehinin in mouse. Methods： Online SPE - HPLC systems were applied on determination of bavachinin in mouse plasma. The chromatographic conditions were as follows ： a C18 column （ Venusil MP, 150 mm × 4.6 mm, 5 μm） was adopted with a linear gradient elution, and the mobile phase consisted of acetonitrile -5 mmol ·L-1 monopotassium phosphate buffer at a flow rate of 1.0 mL ·min-1 ;a MF Ph - 1 online SPE column （ 10 mm × 4 mm, 5 μm） was adopted with isocratic elution, and the washing solvent was 5 mmol · L^-1 monopotassium phosphate buffer;the detected wavelength was set at 238 nm. The pharmaeokinetic parameters were calculated by WinNonlin 5.2 software. Results： The linear range of the calibration curve was 20.0 - 4000 ng · mL- 1, and the lower limit of qualification was 20 ng · mL- 1. The extraction recovery was 90. 5% -94.6%. The RSD of intra - day and inter - day precision was less than 10%. The accuracy of short - term stability, freeze - thaw stability and long - term stability was from 92. 38% to 101.7%. Conclusion：The method is proved to be simple, rapid and high efficiency, and the result is reliable and reproducible.