[关键词]
[摘要]
目的 针对炙黄芪中主要指标成分及经蜜炙转化的特征乙酰基成分,建立HPLC-PDA-ELSD测定炙黄芪中6种异黄酮及2种三萜皂苷的分析方法以及6种异黄酮的一测多评(quantitative determination analysis multi-component by a single-marker,QAMS)法。方法 选取毛蕊异黄酮7-O-β-D-葡萄糖苷(CYG)、黄芪皂苷I(AGI)分别为异黄酮和三萜皂苷类成分的内参物,并计算CYG与乙酰毛蕊异黄酮苷(CYA)、芒柄花苷(FMG)、毛蕊异黄酮(CY)、乙酰芒柄花苷(FMA)、芒柄花素(FM)及AGI与乙酰黄芪皂苷I(ATI)的相对校正因子(fk/s),在不同仪器、色谱柱、柱温、体积流量上考察各成分相对校正因子的稳定性与耐用性,比较QAMS法与外标法(ESM)结果差异。结果 对来自8个厂家共18批炙黄芪进行检测,PDA检测的6种异黄酮的QAMS法与ESM的结果相近,相对误差绝对值均在5%以内,表明针对异黄酮类成分建立的QAMS法准确性良好,可选用QAMS法与ESM进行质量控制;ELSD检测的2种三萜皂苷的QAMS法与ESM的结果差异明显,相对误差绝对值为0.40%~22.69%,显示针对三萜皂苷类成分建立的QAMS法不适用,应选用ESM进行质量控制。结论 建立的炙黄芪中6种异黄酮及2种三萜皂苷的分析方法及6种异黄酮的QAMS法准确可靠,操作简便,可反映黄芪的蜜炙程度,为更加全面地评价炙黄芪质量提供参考,以期提升炙黄芪饮片质量。
[Key word]
[Abstract]
Objective Aiming at the major index components and characteristic acetyl components of Huangqi (Astragali Radix, AR) processed by honey, an analytical method by HPLC-PDA-ELSD for the determination of six isoflavones and two triterpenoid saponins as well as a quantitative determination analysis multi-component by a single-marker (QAMS) method for six isoflavones in honey-processed Astragali Radix were established. Methods Calycosin-7-O-β-D-glucoside (CYG) and astragaloside I (AGI) were selected separately as internal reference substances of isoflavones including calycosin-7-O-glucoside-6ʺ-O-acetyl (CYA), formononetin-7-O-glucoside (FMG), calycosin (CY), formononetin-7-O-glucoside-6ʺ-O-acetyl (FMA), formononetin (FM) and triterponid saponins including acetylastragaloside I (ATI), and relative correction factors (RCFs) between each component and internal reference were calculated. The stability and durability of the measured RCFs were investigated on different instruments, chromatographic columns, column temperatures and flow rates. The deviation of results of QAMS method and the external standard method (ESM) were compared. Results A total of 18 batches of honey-processed Astragali Radix from eight manufacturers were detected of which the QAMS method and ESM results of six isoflavones detected by PDA were similar and the absolute values of relative error were within 5%, indicating that the accuracy of the established QAMS method was good, thus the QAMS method and ESM could be used for quality control about isoflavones. There were larger differences between the QAMS method and ESM results of two triterponid saponins detected by ELSD, of which the absolute values of relative error ranged from 0.40% to 22.69%, indicating that the QAMS method established for triterpenoid saponins was not applicable, thus ESM should be selected for quality control about triterpenoid saponins. Conclusion The established analytical method for the determination of six isoflavones and two triterpenoid saponins and the QAMS method for the determination of six isoflavones in honey-processed Astragali Radix were accurate, reliable and easy to operate, which can reflect the degree of honey-processed Astragali Radix and provide a reference for the quality control of honey-processed Astragali Radix more comprehensively, in order to improve the quality of honey-processed Astragali Radix decoction pieces.
[中图分类号]
R286.2
[基金项目]
国家自然科学基金项目(81973482)