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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 investigate the application of on-line solid-phase extraction (SPE) -high performance liquid chromatography (HPLC) system in the pharmacokinetics of psoralen in mouse plasma. Methods: The serum concentration of psoralen was determined by Ultimate 3000 system based on online SPE-HPLC technique. Chromatographic conditions: Venusil MPC18 (150 mm × 4.6 mm, 5 μm) was used. The mobile phase consisted of acetonitrile-5 mmol·L-1 potassium dihydrogen phosphate buffer (pH 3.5), flow rate 1.0 mL · min-1 ; Using Capcell MF Ph-1 on-line SPE column (10 mm × 4 mm, 5 μm) with 5 mmol·L-1 potassium dihydrogen phosphate buffer (pH 3.5) as the eluent; detection wavelength was 238 nm. WinNonlin 5.2 software calculates pharmacokinetic parameters. Results: The linear range of psoralen was 20.0-4000 ng · mL-1, the lowest limit of quantification was 20.0 ng · mL-1 and the recovery rate was 90.5% -94.6%. The intra-day and inter-day precision test RSD were less than 10%, short-term stability, freeze-thaw stability and long-term stability of the accuracy of 92.38% ~ 101.7%. Conclusion: This method has the advantages of simplified workflow, rapid sample analysis and improved work efficiency. The results are credible and reproducible.