HPLC法测定大鼠尿液中瑞香素的浓度

目的: 建立简单、灵敏、准确的大鼠尿液中瑞香素浓度的HPLC测定方法。 方法: 尿样经乙酸乙酯萃取,采用HPLC法检测,色谱柱为Agilent Zorbax SB-C₁₈(250 mm×4.6 mm,5 μm)柱,流动相为甲醇-0.5%醋酸水(22: 78),流速1 mL·min^-1,检测波长327 nm。大鼠灌胃不同剂量的瑞香素固体分散体溶液(5,25,125 mg·kg^-1),收集0~2,2~6,6~12,12~24,24~48 h的尿液并分析,计算瑞香素在尿液中的累积排泄率。 结果: 瑞香素尿液的线性范围为0.155~396 μg·mL^-1(r=0.9991),定量限为0.155 μg·mL^-1。大鼠灌胃给药瑞香素后2 h即能检测到原型药物,同时发现其代谢产物,24 h内经尿液排泄基本完全,累积排泄率不到6%。 结论: 所建立的方法准确可靠,可用于大鼠灌胃瑞香素后原型药物尿样浓度的测定及其尿排泄研究。

Objective: To develop a rapid and sensitive HPLC method to determine daphnetin in rat urine. Methods: The analytes were extracted with ethyl acetate.The separation was performed on an Agilent Zorbax SB-C₁₈ column(250 mm×4.6 mm,5 μm) with methanol-0.5% acetic acid(22: 78) as mobile phase at a flow rate of 1 mL·min^-1 and a wavelength of 327 nm.Solid dispersion solution of daphnetin was administered intragastrically to rats at doses of 5,25 and 125 mg·kg^-1,and the cumulative excretion rate of daphnetin in urine were determined at different time(0-2,2-6,6-12,12-24,24-48 h). Results: The calibration curve was linear in the range of 0.155-396 μg·mL^-1(r=0.9991) with LOQ of 0.155 μg·mL^-1.Daphnetin and its metabolites were detected in the urine collected within 2 h after intragastric administration and almost completely excreted within 24 h.Cumulative urinary excretion of 24 h was not more than 6% of administration dosage. Conclusion: The method is accurate,reliable and suitable for determination of daphnetin in rat urine and the study of cumulative excretion.

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