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,5 μm),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(0.05,5,25 μg/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 11m and the column temperature was maintained at 35℃. The sample injection volume was 20 μL. 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. The 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 fol-lows: Cmax(2.033±0.41 )μg/mL, tmax(0. 5) h, r1/2ke(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.
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