HPLC波长切换法同时测定杜仲双降袋泡剂中7种成分的含量

Simultaneous determination of seven ingredients in Duzhong Shuangjiang Daipaoji by HPLC

目的建立高效液相(HPLC)波长切换法同时测定杜仲双降袋泡剂中桃叶珊瑚苷、京尼平苷酸、绿原酸、京尼平苷、松脂醇二葡萄糖苷、芦丁及槲皮素7种成分含量的方法。方法以80%甲醇为溶剂,加热回流提取;色谱柱:Hydrosphere C18(4.6 mm×200 mm,5μm);柱温:30℃;流动相:乙腈-0.1%磷酸溶液,梯度洗脱;检测波长:208 nm(桃叶珊瑚苷)、235 nm(京尼平苷酸、绿原酸、京尼平苷、松脂醇二葡萄糖苷)、270 nm(芦丁、槲皮素);流速:0.9 ml/min。结果桃叶珊瑚苷、京尼平苷酸、绿原酸、京尼平苷、松脂醇二葡萄糖苷、芦丁、槲皮素的质量浓度分别在10.97~274.25μg/ml(r=0.999 7)、9.78~244.50μg/ml(r=0.999 8)、6.86~171.50μg/ml(r=0.999 6)、2.47~61.75μg/ml(r=0.999 7)、8.11~202.75μg/ml(r=0.999 1)、4.59~114.75μg/ml(r=0.999 9)、1.85~46.25μg/ml(r=0.999 5)范围内线性关系良好;平均加样回收率分别为99.85%、98.92%、97.52%、97.08%、98.51%、97.10%、96.91%,RSD分别为0.93%、0.74%、1.26%、1.37%、0.88%、1.05%、1.33%。结论所建立的HPLC波长切换法可以同时测定杜仲双降袋泡剂中桃叶珊瑚苷、京尼平苷酸、绿原酸、京尼平苷、松脂醇二葡萄糖苷、芦丁和槲皮素的含量,方法简便准确、灵敏度高,可用于杜仲双降袋泡剂的质量控制。

Objective To establish an HPLC method for the simultaneous determination of the contents of aucubin,geniposidic acid,chlorogenic acid,geniposide,pinoresinol diglucoside,rutin and quercetin in Duzhong Shuangjiang Daipaoji.Methods Samples were prepared by reflux extraction with 80%methanol.The chromatography method was accomplished on an Hydrosphere C18(4.6 mm×200 mm,5μm)chromatographic column,with a mobile phase consisting of acetonitrile-0.1%phosphoric acid solution with gradient elution;the column temperature was set at 30℃;aucubin was detected at 208 nm;geniposidic acid,chlorogenic acid,geniposide and pinoresinol diglucoside were detected at 235 nm;rutin and quercetin were detected at 270 nm;the flow rate was 0.9 ml/min.Results The linear correlation of aucubin,geniposidic acid,chlorogenic acid,geniposide,pinoresinol diglucoside,rutin and quercetin showed good linearity in the given concentration range of 10.97~274.25μg/ml(r=0.9997),9.78~244.50μg/ml(r=0.9998),6.86~171.50μg/ml(r=0.9996),2.47~61.75μg/ml(r=0.9997),8.11~202.75μg/ml(r=0.9991),4.59~114.75μg/ml(r=0.9999)and 1.85~46.25μg/ml(r=0.9995).The average recoveries and RSDs were 99.85%(0.93%),98.92%(0.74%),97.52%(1.26%),97.08%(1.37%),98.51%(0.88%),97.10%(1.05%)and 96.91%(1.33%),respectively.Conclusion This HPLC wavelength switching method could simultaneously determin the contents of aucubin,geniposidic acid,chlorogenic acid,geniposide,pinoresinol diglucoside,rutin and quercetin in Duzhong Shuangjiang Daipaoji.The method is simple,accurate and sensitive,which can be used for the quality control of Duzhong Shuangjiang Daipaoji.