大鼠体内槲皮素的血药浓度测定及其药代动力学研究

Study on Plasma Concentration and Pharmacokinetics of Quercetin in Rats by HPLC

目的建立测定大鼠血浆中槲皮素浓度的高效液相色谱法,并应用于药代动力学研究。方法10只大鼠(雌雄各半)灌胃给予槲皮素50 mg/kg。采用高效液相色谱法测定槲皮素的血药浓度,以山柰酚为内标,流动相为甲醇-缓冲液(0.1 mol/L NH4AC+0.3 mmol/LEDTA-Na2)-乙酸=(60∶40∶1,V/V),色谱柱为迪马ODS C18柱(250 mm×4.6 mm,5μm),测定波长为370 nm,进样量为20μL,流速为1 mL/min,柱温为35℃。结果槲皮素血药浓度线性范围是0.01～50μg/mL(r=0.999 9),定量下限为0.01μg/mL。低、中、高(0.05,5,25μg/mL)3个质量浓度的回收率为97.2%～103.4%,日内、日间精密度的RSD均小于10%。大鼠灌胃给予槲皮素50 mg/kg后的最大血药浓度为(2.033±0.41)μg/mL,达峰时间为0.5 h,t1/2ke为(4.17±0.64)h,0-∞药时曲线下面积为(6.77±0.80)μg/(h.mL)。结论方法专属性高,准确、灵敏,可为今后槲皮素的药代动力学研究提供方法学依据。

Objective To develop a HPLC method for the determination of quercetin in rat plasma for the subsequent pharmacokinetic study. Methods 10 rats（5 males and 5 females ） were given quercetin 50 mg/kg by gastric gavage. The HPLC method was used for determining the quercetin concentration in rat plasma,using kaempferol as the internal standard. The chromatographic column was the Diamonsil C18 column （250 mm ×4. 6 mm,5 μm）. The mobile phase consisted of methanol-buffer solution （0. 1 mol/L NH4AC ＋ 0. 3 mmol/L ED- TA - Na2） - acetic acid （60 ： 40 ：1, V/ V）. The flow rate was 1 mL/min. The UV detector was set at 370 nm and the column temperature was maintained at 35 ℃. The sample injection volume was 20 I.tL. Results The linear relationship was in the range of 0. 01 -50 μg/mL for quercetin （r=0. 999 9）.The lowest detection limit was 0. 01 μg/mL. Tbe recovery rate in （0. 05,5,25 μg/mL） 3 concentrations was 97.2% -103.4% and RSD of intra-day and inter-day was less than 10%. The mean pharmacokinetic parameters for rats were as follows： Cmax（2. 033 ±0.41） μg/mL, tmax（0. 5） h, t1/2ke（4. 17 ±0. 64） h, AUC0-∞ （6. 77 ±0. 80） μg/（h · mL）. Conclusion This method has good specificity,good accuracy and high sensitivity,which can provide the methodological basis for further study of quercetin pharmacokinetics.