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目的:建立HPLC法同时测定大鼠血浆中的橙皮素和柚皮素。方法:采用C_(18)色谱柱,甲醇和磷酸盐缓冲液为流动相,线性梯度洗脱,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,加入二氯甲烷使有机相和水相分层,取有机相氮气吹干后复溶。结果:橙皮素和柚皮素均在0.05～25μg·ml~(-1)浓度范围内线性关系良好,回收率分别为93.7%～113.2%和97.7%～104.3%,日内、日间RSD分别小于11.3%和13.9%。结论:该法专属性强,准确可靠,可用于橙皮素和柚皮素的药物动力学研究。
Objective: To establish a HPLC method for simultaneous determination of hesperetin and naringenin in rat plasma. Methods: A C 18 column was used. The methanol and phosphate buffers were used as the mobile phase. The gradient elution was linear and the detection wavelength was 288 nm. After the plasma sample was hydrolyzed by glucuronidase and sulfatase, the protein was precipitated with ethanol, dichloromethane was added to separate the organic phase from the aqueous phase, and the organic phase was re-dissolved after drying with nitrogen. RESULTS: The linear relationship between hesperetin and naringenin was in the range of 0.05 to 25 μg·ml -1 . The recoveries were 93.7% to 113.2% and 97.7% to 104.3%, respectively. Less than 11.3% and 13.9%. Conclusion: This method is highly specific, accurate and reliable, and can be used for pharmacokinetic studies of hesperetin and naringenin.