高效液相色谱法测定大鼠血浆中总槲皮素浓度及其药动学的应用

目的：建立高效液相色谱法测定大鼠血浆中总槲皮素的浓度，并应用于其药动学研究。方法：血浆样本在酸性条件下经酶水解后，经乙酸乙酯进行萃取，以乙腈-0.2%磷酸水溶液（30：70）为流动相，阿魏酸作内标，采用Inertsil ODS-SP（4.6 mm×150 mm，5 μm）色谱柱进行分离，检测波长366 nm，进样体积20 μL，流速1.0 mL·min^-1，柱温30℃。8只SD雄性大鼠单次灌胃槲皮素50 mg·kg^-1后采集血样，以高效液相色谱法测定槲皮素浓度，采用DAS3.0软件计算药动学参数并进行分析。结果：槲皮素在0.10~10.07 μg·mL^-1范围内线性关系良好，方法定量下限为0.10 μg·mL^-1，回收率为88.85%~101.66%，日内精密度为1.11%~2.79%，日间精密度为2.60%~3.64%。大鼠单次灌胃50 mg·kg^-1的槲皮素后的C_max=（1.97±0.41） μg·mL^-1，t_1/2=（6.10±2.84） h，t_max=（8.63±3.89）h，AUC_0-∞=（29.07±8.24） μg·h·mL^-1。结论：该方法专属性强、灵敏度高，能准确地定量大鼠血浆中槲皮素的总浓度，适用于槲皮素的药代动力学研究。

Determination of total quercetin in rat plasma by HPLC and its application in pharmacokinetics study

OBJECTIVE To develop a HPLC method for the determination of quercetin in the plasma of rats.METHODS The acidified sample was extracted by ethyl acetate. The analytical column was Inertsil ODS-SP (4.6×150 mm, 5 μm). The mobile phase was composed of acetonitrile-0.2% phosphoric acid aqueous solution (30:70) and the internal standard was ferulic acid. The flow rate was 1.0 mL·min^-1. The UV detector was set at 366 nm and the column temperature was maintained at 30℃. The sample injection volume was 20 μL. The pharmacokinetic characteristics of quercetin were investigated in 8 male SD rats. After single oral dose administration of quercetin (50 mg·kg^-1), the serial blood samples were collected and analyzed by HPLC established.RESULTS Quercetin was linear in the range of 0.10-10.07 μg·mL^-1 and the lowest detection limit was 0.01 μg·mL^-1. The recovery range of method was from 88.85% to 101.66%. The intra-day precision was 1.11%-2.79%, and the inter-day precision was 2.60%-3.64%. After the rats received a single oral dose of quercetin (50 mg·kg^-1), the main pharmacokinetic parameters were as follows:C_max=(1.97±0.41) μg·mL^-1, t_1/2=(6.10±2.84) h, t_max=(8.63±3.89) h, AUC_0-∞=(29.07±8.24) μg·h·mL^-1. CONCLUSION This method has high specificity and sensitivity, can accurately quantify total concentration of quercetin in rats plasma, and is suitable for the pharmacokinetic study of quercetin.