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
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篇名: | 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 |
TITLE: | Content Determination of 8 Components in Astragalus membranaceus before and after Bidirectional Solid Fer- mentation with Cordyceps kyushuensis by HPLC-DAD-ELSD |
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Agilent5TC-C18;流动相为0.1%甲酸水溶液(A)-乙腈(B),梯度洗脱;流速为1mL/min;柱温为30℃;DAD检测波长为260nm;ELSD蒸发管温度为100℃;雾化器温度为80℃;载气流速为1.6L/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。 |
ABSTRACT: | OBJECTIVE:To establish a method for simultaneous determination of calycosin glucoside ,ononin,calycosin, formononetin,astragaloside Ⅳ,isoastragaloside Ⅱ,cycloastragenol and isoastragaloside Ⅰ in 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 15 μL. RESULTS:The eight components had a good linear relationship within their respective ranges of concentration (all R2>0.999 0); 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 astragaloside Ⅳ,isoastragaloside Ⅰ and isoastragaloside Ⅱ can be increased significantly in A. membranaceus . |
期刊: | 2020年第31卷第23期 |
作者: | 杜以晴,于钦辉,文冉,菅彤彤,容蓉,吕青涛,张国英 |
AUTHORS: | DU Yiqing ,YU Qinhui ,WEN Ran,JIAN Tongtong ,RONG Rong,LYU Qingtao ,ZHANG Guoying |
关键字: | 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 |
KEYWORDS: | Astragalus membranaceus ;Cordyceps kyushuensis ;Bidirectional solid fermentation ;HPLC-DAD-ELSD;Content |
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