高效液相色谱法测定大鼠血浆中桂皮酸和马尿酸  

HPLC determination of cinnamic acid and hippuric acid in rat plasma

作　　者：陈莹蓉[1] 马越鸣[1] 张宁[2] 贾伟[3] 李国文[2] 

机构地区：[1]上海中医药大学中药药代动力学实验室,上海201203 [2]上海中医药大学科技实验中心,上海201203 [3]上海中医药大学中医方证与系统生物学研究中心,上海201203

出　　处：《药物分析杂志》2009年第2期212-216,共5页Chinese Journal of Pharmaceutical Analysis

基　　金：上海市重点学科建设项目资助(№T0301)

摘　　要：目的:建立同时测定大鼠血浆中桂皮酸和马尿酸浓度的高效液相色谱法。方法:血浆样品用盐酸和乙酸乙酯萃取,采用SUPELCO Discovery C18柱(5μm,4.6 mm×250 mm),以0.05%磷酸-乙腈为流动相梯度洗脱,流速1.0 mL·min-1;以对羟基苯甲酸甲酯为内标,在230 nm(马尿酸)和278 nm(桂皮酸)波长下进行检测。结果:桂皮酸的线性范围为0.05～5.0μg·mL-1(r=0.9987),最低定量限为0.05μg·mL-1,方法回收率为87.5%～102.4%,绝对回收率大于80%,日内和日间的RSD均小于13.1%;马尿酸的线性范围为0.2～20.0μg·mL-1(r=0.9957),最低定量限为0.2μg·mL-1,方法回收率为96.2%～101.6%,绝对回收率大于70%,日内和日间的RSD均小于8.4%。血浆样品的稳定性符合要求。结论:所建立的大鼠血浆中桂皮酸和马尿酸浓度的HPLC测定方法灵敏、准确、精密、稳定,并适用于大鼠体内桂枝的药代动力学研究。Objective：To develop a method for the determination of cinnamic acid and hippuric acid in rat plasma by HPLC. Methods：The separation was carried out by using a SUPELCO Discovery column （5 μm ,4.6 mm×250 mm） with a gradient mobile phase of 0. 05% phosphoric acid -acetonitrile, at a flow rate of 1.0 mL·min^-1,and detection wavelengths were set at 230 nm （hippuric acid） and 278 nm （cinnamic acid）. Plasma samples were extracted with hydrochloric acid and ethyl acetate and methyl parahydroxybenzoate was used as an internal standard. Results ： The linear range of cinnamic acid was 0. 05 to 5.0 μg·mL^-1 （r=0.9987） with the lower limit of quanti- fication for 0.05 μg·mL^-1.The method recovery was 87.5% to 102.4% and the absolute recovery was more than 80%. The intra -day RSD and inter -day RSD were less than 13. 1%. For hippuric acid,the linear range was 0. 2 to 20. 0 μg·mL^-1（r=0.9957） with the lower limit of quantification for 0. 2 μg·mL^-1. The method recovery was 96.2% to 101.6% and the absolute recovery was more than 70%. The intra-day RSD and inter-day RSD were less than 8.4%. Conclusion：The developed method for the quantification of cinnamic acid and hippuric acid in rat plasma is sensitive, accurate, precise, stable, and is suitable for the pharmacokinetic studies of Ramulus Cinnamomi in rats.

关 键 词：桂皮酸 马尿酸 桂枝 药代动力学 高效液相色谱法 

分 类 号：R917[医药卫生—药物分析学]