HPLC-ECD法测定大鼠血浆中银杏叶黄酮成分及药代动力学研究  

HPLC-ECD determination of flavonols from Ginkgo biloba leaf extracts in rat plasma and its pharmacokinetic study

作　　者：张颖[1] 刘建勋[1] 林力[1] 张强强[1] 王建农[1] 

机构地区：[1]中国中医科学院西苑医院实验研究中心,北京100091

出　　处：《药物分析杂志》2011年第1期10-14,共5页Chinese Journal of Pharmaceutical Analysis

基　　金：科技部重大新药创制专项(20092x09502-014);国家自然科学基金重点项目(30830118)

摘　　要：目的：建立大鼠灌胃银杏叶提取物后血浆中黄酮类成分的灵敏分析方法,并进行药代动力学研究。方法：血浆样品经葡萄糖醛酸苷水解酶水解后,采用高效液相色谱分离及电化学检测,同时分析大鼠血浆样品中槲皮素、异鼠李素和山柰酚。色谱柱为Agilent Zorbax Ec lipse XDB-C8（150 mm×4.6 mm,5μm）,流动相为甲醇-异丙醇-磷酸盐缓冲液（磷酸二氢钠-磷酸,pH=2）（15？15？70）,流速0.8 mL.m in-1,检测电压为200 mV,柱温30℃。结果：槲皮素、异鼠李素和山柰酚与其他内源性和药源性成分达到良好分离;最低检测浓度达到0.5 ng.mL-1;平均提取回收率分别为88.7%,86.2%,87.0%;准确度在92.7%~105.3%之间,日内精密度RSD小于6.6%,日间精密度RSD小于14.1%,样品分析时间为20 m in。大鼠灌胃给予银杏叶提取物10 mg.kg-1后,酶水解血浆中槲皮素、异鼠李素和山柰酚的半衰期分别为（3.53±1.88）h,（6.94±4.05）h,（3.97±2.30）h;3个黄酮类成分在大鼠血浆中都显示明显的二次达峰现象。结论：建立的检测方法能灵敏、特异、准确,可用于银杏叶提取物灌胃给药后黄酮类成分的药代动力学研究。Objective：To establish a sensitive method for the quantification of flavonols from rat plasma after intragastric administration of Ginkgo biloba leaf extracts.Methods：HPLC with electrochemical detection（ECD） was employed to quantify quercetin,isorhamnetin and kaempferol simultaneously after plasma samples were hydrolyzed by glucuronidase.The sample was separated by Agilent Zorbax Eclipse XDB-C8（150 mm×4.6 mm,5 μm） column and eluted by the mobile phase composed of methanol-isopropanol-phosphate buffer（NaH2PO4-H3PO4,pH=2）（15∶15∶70） at column temperature of 30 ℃,and the flow rate was 0.8 mL·min-1.The potential of analysis cell was selected as 200 mV.Results：The analysis of plasma samples could be performed in 20 min with the quantitative limit of 0.5 ng·mL-1 and average recovery of 88.7%,86.2% and 87.0% for quercetin,isorhamnetin and kaempferol,respectively.The intra-and inter-day precision（RSD） was measured less than 6.6% and 14.1%,meanwhile the accuracy was calculated as 92.7%-105.3%.The concentration-time data after oral administration of 10 mg·kg-1 Ginkgo biloba leaf extracts showed that flavanols was absorbed rapidly and presenting the fist peak appeared at 0.17-0.23 h and the second peak at 6 h.The elimination half-life time（t1/2k） was calculated as（3.53±1.88） h,（6.94±4.05） h and（3.97±2.30） h for quercetin,isorhamnetin and kaempferol,respectively.Conclusion：The method developed in the present study using HPLC-ECD is highly sensitive,accurate,rapid and suitable for pharmacokinetic study of Ginkgo biloba leaf extracts in rats.

关 键 词：高效液相色谱-电化学 银杏叶提取物 黄酮 血药浓度 药代动力学 

分 类 号：R917[医药卫生—药物分析学]