高效液相色谱法同时测定醉鱼草中五种黄酮成分

Simultaneous determination of five kinds of flavonoid ingredients in Buddleia by HPLC

目的:建立同时测定中药材醉鱼草中黄酮成分蒙花苷、芦丁、木犀草素、槲皮素和芹菜素含量的方法,为醉鱼草的质量控制提供理论依据。方法:采用高效液相色谱法对醉鱼草中五种黄酮类有效成分进行同时测定。色谱柱型号:Hypersil ODS色谱柱(200 mm×4.6 mm,5μm),柱温:25℃;流速:1.0mL·mL^(-1);流动相:甲醇-0.2%磷酸水溶液;检测波长:256 nm,梯度洗脱。结果:中药材醉鱼草中的黄酮成分蒙花苷、芦丁、木犀草素、槲皮素和芹菜素在质量浓度为1.63~75μg·mL^(-1)(r=0.999 5),0.42~26μg·mL^(-1)(r=0.999 7),0.77~152μg·mL^(-1)(r=0.999 4),1.02~108μg·mL^(-1)(r=0.999 6),0.91~99μg·mL^(-1)(r=0.999 2)范围内与峰面积线性相关,醉鱼草样品加标回收率均值分别为99.7%,97.9%,99.7%,99.5%,99.1%,RSD分别为1.3%,1.7%,0.7%,1.1%,1.0%。结论:该测定方法适用性强、快捷,重复性好,适用于中药材醉鱼草的质量控制。

OBJECTIVE To establish a method for simultaneous determination of flavonoids including buddleoside,rutin,luteolin,quercetin and apigenin in Chinese herbal medicine Buddleja,and provide the theoretical basis for the quality control of Buddleja.METHODS Based on HPLC method,Hypersil ODS chromatographic column（200 mm×4.6 mm,5μm）was used and the mobile phase was composed of methyl alcohol（A）-0.2% phosphoric acid solution（B）.Flavonoid was determined at 1.0 mL·min-1,25 ℃,256 nm wavelength by gradient elution.RESULTS The calibration curves were linear with in the range of1.63-75μg·mL-1（r=0.999 5）for buddleoside,0.42-26μg·mL-1（r=0.999 7）for rutin,0.77-152μg·mL-1（r=0.999 4）for luteolin,1.02-108μg·mL-1（r=0.999 6）for quercetin and 0.91-99μg·mL-1（r=0.999 2）for apigenin,the average recovery rates of Buddleja of buddleoside,rutin,luteolin,quercetin and apigenin were 99.7%,97.9%,99.7%,99.5% and 99.1%,the RSD values were 1.3%,1.7%,0.7%,1.1% and 1.0%.CONCLUSION The method has the advantages of strong applicability,fastness and good reproducibility,and is suitable for the quality control of Chinese herbal medicine Buddleja.