采用反相高效液相色谱-紫外可见检测法同时分离并测定大鼠尿液和鸡血浆中的六种黄酮类化合物及曲克芦丁。

The simultaneous separation and determination of six flavonoids and troxerutin in rat urine and chicken plasma by reversed-phase high-performance liquid chromatography with ultraviolet-visible detection.

作者信息

Yang Gong-Jun, Liu Ping, Qu Xi-Long, Xu Min-Juan, Qu Qi-Shu, Wang Cheng-Yin, Hu Xiao-Ya, Wang Zhi-Yue

机构信息

College of Chemistry and Chemical Engineering, Yangzhou University, Yangzhou, 225002, PR China.

出版信息

J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Sep 1;856(1-2):222-8. doi: 10.1016/j.jchromb.2007.06.002. Epub 2007 Jun 12.

DOI:10.1016/j.jchromb.2007.06.002

PMID:17588830

Abstract

The method of high-performance liquid chromatography (HPLC) with UV-vis detection was used and validated for the simultaneous determination of six flavonoids (puerarin, rutin, morin, luteolin, quercetin, kaempferol) and troxerutin in rat urine and chicken plasma. Chromatographic separation was performed using a VP-ODS column (150 mm x 4.6 mm, 5.0 microm) maintained at 35.0 degrees C. The mobile phase was a mixture of water, methanol and acetic acid (57:43:1, v/v/v, pH 3.0) at the flow rate of 0.8 mL/min. Six flavonoids and troxerutin were analyzed simultaneously with good separation. On optimum conditions, calibration curves were found to be linear with the ranges of 0.10-70.00 microg/mL (puerarin, rutin, morin, luteolin, quercetin, kaempferol) and 0.50-350.00 microg/mL (troxerutin). The detection limits were 0.010-0.050 microg/mL. The method was validated for accuracy and precision, and it was successfully applied to determine drug concentrations in rat urine and chicken plasma samples from rat and chicken that had been orally administered with six flavonoids and troxerutin.

摘要

采用高效液相色谱（HPLC）-紫外可见检测法，对大鼠尿液和鸡血浆中6种黄酮类化合物（葛根素、芦丁、桑色素、木犀草素、槲皮素、山奈酚）和曲克芦丁进行了同时测定，并对该方法进行了验证。色谱分离采用VP-ODS柱（150 mm×4.6 mm，5.0μm），柱温35.0℃。流动相为水、甲醇和乙酸的混合溶液（57:43:1，v/v/v，pH 3.0），流速为0.8 mL/min。6种黄酮类化合物和曲克芦丁同时得到良好分离。在最佳条件下，葛根素、芦丁、桑色素、木犀草素、槲皮素、山奈酚的校准曲线在0.10 - 70.00μg/mL范围内呈线性，曲克芦丁的校准曲线在0.50 - 350.00μg/mL范围内呈线性。检测限为0.010 - 0.050μg/mL。该方法的准确性和精密度得到验证，并成功应用于测定口服6种黄酮类化合物和曲克芦丁的大鼠和鸡的尿液及血浆样品中的药物浓度。