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目的建立大鼠尿中咖啡酸、绿原酸、1,3-二咖啡酰奎宁酸浓度的高效液相色谱测定方法,研究3个化合物在大鼠尿中的排泄情况。方法给大鼠静脉注射灯盏细辛注射液后,于不同时间段采集尿样,尿样中加入内标物原儿茶醛,经乙酸乙酯处理后,样品进HPLC分析。结果尿样中咖啡酸、绿原酸、1,3-二咖啡酰奎宁酸的方法回收率分别约为95%~98%、90%~95%、95%~97%,60 h尿中咖啡酸、绿原酸、1,3-二咖啡酰奎宁酸的累积排泄量占给药剂量的(70.97±7.31)%、(87.62±12.23)%、(9.87±2.63)%。日内及日间精密度均小于10%。结论该方法简便、准确、专属性强,可用于大鼠尿液中3个化合物的含量测定。
OBJECTIVE To establish a method for the determination of urinary caffeic acid, chlorogenic acid and 1,3-dicaffeoylquinic acid in rat urine by high performance liquid chromatography (HPLC), and to study the excretion of three compounds in rat urine. Methods After intravenous injection of Erigeron Breviscapus Injection to rats, urine samples were collected at different time points. Protocatechuic aldehyde, an internal standard, was added into the urine samples. After being treated with ethyl acetate, the samples were analyzed by HPLC. Results The recoveries of caffeic acid, chlorogenic acid and 1,3-dicaffeoylquinic acid in urine were 95% ~ 98%, 90% ~ 95%, 95% ~ 97% The cumulative excretion of caffeic acid, chlorogenic acid and 1,3-dicaffeoylquinic acid accounted for (70.97 ± 7.31)%, (87.62 ± 12.23)% and (9.87 ± 2.63)%, respectively. Intra- and inter-day precision are less than 10%. Conclusion The method is simple, accurate and specific. It can be used for the determination of three compounds in urine of rats.