HPLC法同时测定大鼠胆汁中二苯乙烯苷及其代谢物的浓度  

HPLC simultaneous determination of stilbene glycoside and its metabolite in rat bile liquid

作　　者：王春英[1] 袁志芳[1] 王巧[1] 谢凤立[1] 杨维[1] 张兰桐[1] 

机构地区：[1]河北医科大学药学院药物分析教研室,石家庄050017

出　　处：《药物分析杂志》2009年第1期53-57,共5页Chinese Journal of Pharmaceutical Analysis

基　　金：河北省自然基金资助项目(C2006000791)

摘　　要：目的:建立大鼠胆汁中二苯乙烯苷及其代谢物浓度的HPLC测定方法,明确二苯乙烯苷的代谢途径。方法:胆汁样品或用重蒸水稀释后的胆汁样品200μL,加入甲醇10μL后直接离心测定二苯乙烯苷及其代谢物的浓度。Dikma Diamonsil C18色谱柱(250mm×4.6mm,5μm),柱温30℃;流动相为乙腈-0.1%醋酸梯度洗脱(0～10min,15:85;15～23min,25:75;25～30min,100:0),流速1mL.min-1;检测波长320nm。测定大鼠灌胃给予二苯乙烯苷60mg.kg-1后胆汁中二苯乙烯苷及其代谢物的累积排泄率。结果:二苯乙烯苷及其代谢物在胆汁中的线性范围分别为12.0～0.8μg.mL-1和110～0.9μg.mL-1,日内和日间RSD均小于7.0%,准确度在±4.5%之间。提取回收率均大于99.1%。大鼠灌胃给予二苯乙烯苷60mg.kg-1后24h胆汁中二苯乙烯苷及其代谢物的累积排泄率分别为(0.084±0.04)%和(36.87±12.94)%。结论:经方法学考察符合生物样品的测定要求,可应用于大鼠胆汁中二苯乙烯苷及其代谢物浓度的测定,二苯乙烯苷在大鼠体内主要以代谢物的形式由胆汁排泄。Objective ： To develop an HPLC method to assess the concentrations of stilbene glycoside and its metabolite in rat bile and identify the metabolic pathway of stilbene glycoside in rat. Methods： After a single oral administration at a dose of 60 mg · kg^-1 ,the bile of rat was collected. After methanol（ 10 μL） was added into 200 μL bile or diluted bile, and then centrifuged at a high speed for 10 min, the supernatant was injected into the liquid chromatography system to determine the concentrations of stilbene glycoside and its metabolite. Dikma Diamonsil C18 column（250 mm×4. 6 mm,5 μm） was used. The mobile phase was composed of acetonitrile and 0. 1% （v/v） acetic acid solution（0 -10 min, 15：85;15 -23 min,25：75 ;25 -30 min, 100： 0）. The flow rate was 1.0 mL · min^-1 and the column compartment was kept at the temperature of 30 ℃, 320 nm was chosen to record chromatograms. Results：Calibration curves were generated over a concentration range of 12. 0 -0. 8 μg · mL^-1 for stilbene glycoside and 110 -0. 9 μg ·mL^-1 for the metabolite, respectively. Intra - and inter - day RSDs were less than 7. 0%, and relative errors were ranged within ±4. 5%. Extraction recoveries of stilbene glycoside and metabolite were all above 99. 1%. Mean biliary accumulated excretion of stilbene glycoside and metabolite for 24 h after administration were （0. 084 ± 0. 04） % and （ 36. 87 ± 12. 94 ） %, respectively. Conclusion： This specific, sensitive and precise method is suitable for the determination of stilbene glycoside and its metabolite in rat bile. Biliary excretion should be the main excretion route of stilbene glycoside in the form of its metabolite.

关 键 词：二苯乙烯苷 代谢物 含量测定 胆汁排泄 

分 类 号：R917[医药卫生—药物分析学]