大鼠胆汁中木犀草素和芹菜素HPLC测定方法的建立及应用  

HPLC determination of luteolin and apigenin in rat bile and its application

作　　者：陈婷[1] 蒋惠娣[1] 鲁鑫炎[1] 曾苏[1] 

机构地区：[1]浙江大学药学院药物分析与药物代谢研究室,杭州310058

出　　处：《药物分析杂志》2007年第4期490-493,共4页Chinese Journal of Pharmaceutical Analysis

基　　金：浙江省自然科学基金(Y204379);浙江省经贸委(G20050066)

摘　　要：目的:建立大鼠胆汁中木犀草素和芹菜素浓度测定的HPLC方法,并考察大鼠经口给予菊花提取物(CME)后,木犀草素和芹菜素在胆汁中的排泄情况。方法:胆汁样品经葡萄糖醛酸酶和硫酸酯酶酶解后测定,以SB-C_(18)柱(250mm×4.6mm,5μm)为分离柱,流动相为甲醇-0.2%磷酸水溶液(52∶48,v/v),流速为1.0mL·min^(-1),检测波长为350nm。结果:本法木犀草素、芹菜素分离良好,线性范围分别为0.1745～6.986μg·mL^(-1)(r=0.9998)和0.3693～11.08μg·mL^(-1)(r=0.9999),绝对回收率为84.6%～106.2%,方法回收率为80.1%～106.9%,日内和日间精密度RSD小于14%。大鼠灌胃给药CME后36h内,胆汁中木犀草素和芹菜素的累积排泄量占给药量的2.05%和6.34%。结论:本法可以准确、灵敏地同时测定大鼠胆汁中木犀草素和芹菜素的浓度,两者经胆汁排泄证明存在肝肠循环。Objective：To establish a RP - HPLC method for the determination of luteolin and apigenin in rat bile, and study the biliary excretion of luteolin and apigenin after rat orally administrated Chrysanthemum morifolium extract （CME）. Methods：Bile samples were enzymatic hydrolyzed before analysis by RP -HPLC. SB -C18HPLC col- umn （250 mm ×4. 6 mm,5 μm）was used. The mobile phase was methanol-0. 2% phosphoric acid （52：48,v/v） with the flow rate of 1.0 mL ·min^-1 ;the UV detector was set at 350 nm. Results：A good separation was obtained. The standard curves for luteolin and apigenin in bile were linear over the range of 0. 1745 -6. 986 μg ·min^-1 and 0. 3693 -11.08 μg ·min^-1 ,respectively. The absolute recovery for luteolin and apigenin was 84. 6% to 106. 2% and assay recovery was 80. 1% to 106. 9%. The intra - and inter - day precisions （RSD） for luteolin and apigenin were all less than 14%. The relative biliary excretion rate of intake dose Of luteolin and apigenin during 36 hours after oral administration were 2. 05% and 6. 34%, respectively. Conclusion：The method developed is accurate, sensitive for the determination of luteolin and apigenin in bile. Enterohepatic circulation of luteolin and apigenin associated with double peaks in the blood concentration -time curves can be proved by the biliary excretion.

关 键 词：木犀草素 芹菜素HPLC 大鼠 胆汁 菊花提取物(CME) 

分 类 号：R917[医药卫生—药物分析学]