UPLC快速测定葛根芩连汤肠外翻囊样品中6个黄酮类成分含量

目的:建立UPLC法同时测定葛根芩连汤肠外翻囊样品中6个黄酮类成分(葛根素、大豆苷、甘草苷、野黄芩苷、黄芩苷、汉黄芩苷)的含量。方法:样品用甲醇提取,采用Agilent Proshell 120 EC-C18(4.6 mm×50 mm,2.7μm)色谱柱,以0.2%醋酸水溶液(A)-甲醇(B)为流动相进行梯度洗脱,流速1 mL.min-1,紫外检测波长270 nm,柱温30℃。结果:葛根素、大豆苷、甘草苷、野黄芩苷、黄芩苷、汉黄芩苷质量浓度分别在0.1870～93.50μg.mL-1(r=0.9999),0.02730～13.65μg.mL-1(r=0.9999),0.02550～12.75μg.mL-1(r=0.9999),0.05580～27.90μg.mL-1(r=0.9998),0.2124～106.2μg.mL-1(r=0.9999),0.09140～45.70μg.mL-1(r=0.9999)范围内有良好的线性关系;日间、日内精密度变异均小于2%;平均回收率(n=9)分别为101.4%,97.88%,99.15%,99.72%,98.50%,101.4%;稳定性良好。结论:所建立的分析方法操作简便、快速、灵敏,结果准确,重复性好,所需样品少,并已成功应用于葛根芩连汤大鼠肠吸收动力学的研究。

UPLC rapid determination for six flavonoids of Gegenqinlian decoction in rat everted gut sacs

Objective:To develop a sensitive and selective ultra performance liquid chromatography(UPLC) method for identification and quantification of six flavonoids components(puerarin,daidzin,liquiritin,scutellarin,baicalin,wogonoside) in rat everted gut sacs.Methods:The method was based on simple liquid-liquid extraction.The separation was performed on a Agilent Proshell 120 EC-C18(4.6 mm×50 mm,2.7 μm) column.The mobile consisted of 0.2% acetic acid aqueous(A) and methanol(B) with a gradient elution at a flow-rate of 1.0 mL·min-1.The detection wavelength was 270 nm and the column temperature was 30 ℃.Results:The linear ranges of puerarin,daidzin,liquiritin,scutellarin,baicalin,wogonoside were 0.1870-93.50 μg·mL-1(r=0.9999),0.02730-13.65 μg·mL-1(r=0.9999),0.02550-12.75 μg·mL-1(r=0.9999),0.05580-27.90 μg·mL-1(r=0.9998),0.2124-106.2 μg·mL-1(r=0.9999),0.09140-45.70 μg·mL-1(r=0.9999),respectively.The intra-and inter-day precisions(RSD) were within 2% for the analytes.The mean recoveries of puerarin,daidzin,liquiritin,scutellarin,baicalin,wogonoside were 101.4%,97.88%,99.15%,99.72%,98.50%,101.4%.Conclusion:The proposed method enables unambiguous identification and quantification for the preliminary pharmacokinetic studies of the compounds(puerarin,daidzin,liquiritin,scutellarin,baicalin,wogonoside),and has been applied to a intestinal absorption kinetics of Gegenqinlian decoction in rats successfully.