高效液相色谱法测定金钱草胶囊中槲皮素的血药浓度

目的建立金钱草胶囊中槲皮素的血药浓度测定方法,为临床应用和药代动力学研究提供技术参考。方法采用高效液相色谱-紫外检测方法测定,色谱柱为Kromasil C8柱(250 mm×4.6 mm,5μm),流动相为甲醇-乙腈-0.4%磷酸溶液(40∶10∶50),流速1 mL/min,柱温30℃,进样量20μL,检测波长360 nm。血样提取方法,以β-葡萄糖苷酸酶水解后,以乙酸乙酯提取,氮气流吹干后复溶进样。结果槲皮素保留时间约为8.8 min,金钱草胶囊中槲皮素的血药浓度线性范围0.08～12μg/mL,最低定量限为0.05μg/mL,检测限为0.02μg/mL。方法平均回收率为97.50%～102.50%,水解提取回收率为76.46%～87.43%,日内精密度RSD为4.18%～6.17%,日间精密度RSD为5.08%～8.63%。结论方法简便,分析速度快,灵敏度较高,重复性好,适用于临床血药浓度测定及药代动力学研究。

Determination of Plasma Quercetin Concentration after Oral Administration of Jinqianchao Capsules by HPLC

Objective To develop an HPLC method for the determination of quercetin concentration in human plasma to provide the technical reference for its pharmacokinetic study and clinical application.Methods The HPLC-UV method was used.The separation was performed on the Kromasil C8 column（250 mm×4.6 mm,5 μm） with the mobile phase of methanol-acetonitrile-0.4%phosphoric acid solution（40 ∶10 ∶50）.The flow rate was 1.0 mL/min and the detection wavelength was 360 nm.The column temperature was 30 ℃and the injection volume was 20 μL.In the blood sample extraction method,after β-glucuronidase hydrolysis,the plasma was extracted with ethyl acetate and dried under nitrogen.The residue was redissolved with mobile phase and injected into the sampler.Results The retention time of quercetin was about 8.8 min.The calibration curve for quercetin was linear in the concentration range of 0.08-12.0 μg/mL.The lowest limit of quantitation（LOQ） for quercetin was 0.05 μg/mL;the limit of detection（LOD） was 0.02 μg/mL.The methodological recovery rate of low,medium and high concentration was 97.50%-102.50%.The hydrolysis extraction recovery rate was 76.46%-87.43%.The relative standard deviations（RSD） for intra-day and inter-day assay were 4.18%-6.17%and 5.08%-8.63%,respectively.Conclusion The established HPLC method is simple,rapid and highly sensitive with good repeatability,sutable for the determination of quercetin concentration in human plasma and its pharmacokinetic study.