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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 an HPLC method for the determination of hesperetin enantiomers in rat plasma. Methods: The mobile phase was Chiralpak AD-H with n-hexane-ethanol-trifluoroacetic acid (20: 80: 0.5) After plasma samples were hydrolyzed by glucuronidase and sulfatase, the protein was precipitated with ethanol, and the organic phase and the aqueous phase were separated by adding dichloromethane, and the organic phase was nitrogen-dried to reconstitute the ethanol. Results: The linear relationship between the enantiomers of hesperetin R and S in the range of 0.1 ~ 20μg · ml ~ (-1) was good. The recovery rates of hesperetin were 86.54% and 84.62%, respectively. The intra-day and interday RSD were less than 4.8 % And 5.1%. Conclusion: This assay is specific, accurate and reliable and can be used to study the pharmacokinetics of hesperetin enantiomers.