HPLC法测定大鼠血浆中橙皮素对映异构体浓度

Determination the Concentration of Hesperetin in Rat Plasma by HPLC

目的:建立HPLC法测定大鼠血浆中的橙皮素对映异构体。方法:采用ChiralpakAD-H色谱柱,正己烷-乙醇-三氟乙酸(20:80:0.5)为流动相,检测波长288 nm。血浆样品经过葡萄糖醛酸酶和硫酸酯酶水解后,用乙醇沉淀蛋白,并加入二氯甲烷使有机相和水相分层,取有机相氮气吹干后乙醇复溶。结果:橙皮素R,S对映异构体在0.1～20μg·ml^(-1)浓度范围内线性关系良好,提取回收率分别为86.54%和84.62%,日内、日间RSD分别小于4.8%和5.1%。结论:该检测方法专属性强,准确可靠,可用于橙皮素对映异构体的药物动力学研究。

Objective ： To establish a stereospecific method for quantitation of the enatiomers of hesperetin in rat plasma. Method： The enatiomers of the hesperetin were separated on the Chiralpak AD-H column. The mobile phase consisted of n-hexane ： ethonoll ： trifluoroacetie acid （20： 80： 0. 5 ） ,separation was carried out isocratically at a flow rat of 0. 8 ml·min^-1 with ultraviolet detection at 288 nm. Plasma sample was preparation with incubating with 13-glucuronidase/sulphatase, precipitate the protein and isolated the analytes with ethanol,then dichlormethane was added to gain layer separated, organic layer separated was dried and reconsituted in ethonol. Resalt： The calibration curves were both linear in the concentration range of 0. 1-20 μg· ml^-1. The recoveries were 86. 54% and 84. 62%, with the intra- and inter-RSDs less than 4. 8 % and 5.1% respectively. Conclusion： The method established for analyzing naringenin, hesperetin in rat plasma is accurate and specific and has been successfully applied to pharmacokinetic studies in rats.