HPLC-DAD-ELSD法测定黄芪经九州虫草双向固体发酵前后8种成分的含量  

Content Determination of 8 Components in Astragalus membranaceus before and after Bidirectional Solid Fermentation with Cordyceps kyushuensis by HPLC-DAD-ELSD

作　　者：杜以晴 于钦辉 文冉 菅彤彤 容蓉[1] 吕青涛[1] 张国英[1] DU Yiqing;YU Qinhui;WEN Ran;JIAN Tongtong;RONG Rong;LYU Qingtao;ZHANG Guoying(School of Pharmacy,Shandong University of TCM,Jinan 250355,China)

机构地区：[1]山东中医药大学药学院,济南250355

出　　处：《中国药房》2020年第23期2841-2846,共6页China Pharmacy

基　　金：国家自然科学基金资助项目(No.21807066)。

摘　　要：目的:建立同时测定黄芪经九州虫草双向固体发酵前后毛蕊异黄酮苷、芒柄花苷、毛蕊异黄酮、芒柄花素、黄芪皂苷Ⅳ、异黄芪皂苷Ⅱ、环黄芪醇、异黄芪皂苷Ⅰ等8种成分含量的方法,并探讨发酵处理对黄芪中上述8种成分含量的影响。方法:采用高效液相色谱-二极管阵列检测器-蒸发光检测器联用技术(HPLC-DAD-ELSD)进行含量测定。色谱柱为Agilent 5 TC-C18;流动相为0.1%甲酸水溶液(A)-乙腈(B),梯度洗脱;流速为1 mL/min;柱温为30℃;DAD检测波长为260 nm;ELSD蒸发管温度为100℃;雾化器温度为80℃;载气流速为1.6 L/min;进样量为15μL。结果:8种成分在各自质量浓度范围内线性关系良好(R2均大于0.9990);精密度、稳定性、重复性试验的RSD均小于3%(n=3或n=6);平均加样回收率为97.88%~101.32%,RSD为1.22%~2.39%(n=6)。以未发酵黄芪中相应成分含量为100%计算,经九州虫草双向固体发酵后,黄芪中毛蕊异黄酮苷、芒柄花苷、毛蕊异黄酮、芒柄花素、黄芪皂苷Ⅳ、异黄芪皂苷Ⅱ、环黄芪醇、异黄芪皂苷Ⅰ的含量变化率分别为-98.51%、-96.41%、-94.74%、-96.40%、289.20%、20.25%、-75.05%、562.46%。结论:黄芪经过九州虫草发酵后,有利于增加其主要活性成分黄芪皂苷Ⅳ、异黄芪皂苷Ⅰ、异黄芪皂苷Ⅱ的含量。OBJECTIVE:To establish a method for simultaneous determination of calycosin glucoside,ononin,calycosin,formononetin,astragalosideⅣ,isoastragalosideⅡ,cycloastragenol and isoastragalosideⅠin Astragalus membranaceus before and after bidirectional solid fermentation with Cordyceps kyushuensis,and to investigate the effects of fermentation on the contents of above 8 components in A.membranaceus.METHODS:HPLC-DAD-ELSD was adopted.The determination was performed on Agilent 5 TC-C18 column with mobile phase consisted of 0.1%formic acid aqueous solution-acetonitrile(gradient elution)at the flow rate of 1 mL/min.The column temperature was set at 30℃.DAD detection wavelength was set at 260 nm,ELSD evaporation tube temperature was 100℃,atomizer temperature was 80℃,carrier gas flow rate was 1.6 L/min;injection volume was 15μL.RESULTS:The eight components had a good linear relationship within their respective ranges of concentration(all R2>0.9990);RSDs of precision,stability and repeatability tests were all less than 3%(n=3 or n=6);the recoveries was 97.88%-101.32%,and RSDs were 1.22%-2.39%(n=6).Setting the content of components in unfermented A.membranaceus as 100%,after bidirectional solid fermentation with C.kyushuensis,the change rates of 8 components were-98.51%,-96.41%,-94.74%,-96.40%,289.20%,20.25%,-75.05%,562.46%,respectively.CONCLUSIONS:After fermentation with C.kyushuensis,the contents of active components as astragalosideⅣ,isoastragalosideⅠand isoastragalosideⅡcan be increased significantly in A.membranaceus.

关 键 词：黄芪 九州虫草 双向固体发酵 高效液相色谱-二极管阵列检测器-蒸发光检测器联用技术 含量测定 

分 类 号：R283[医药卫生—中药学]