一测多评法测定黄芪中异黄酮及苷类成分

Determination of isoflavonoids and glycosides in Astragali Radix by QAMS method

目的采用一测多评(QAMS)法同时测定黄芪中6个异黄酮及苷类成分的含量,并验证方法在黄芪质量评价中应用的可行性与适用性。方法采用HPLC-UV法建立丙二酰毛蕊异黄酮苷(CYM)、芒柄花苷(FMG)、丙二酰芒柄花苷(FMM)、毛蕊异黄酮(CY)及芒柄花素(FM)与内参物毛蕊异黄酮7-O-β-D糖苷(CYG)的相对校正因子(f_(s/i)),并在不同色谱仪和色谱柱上考察各成分相对校正因子的稳定性和耐用性,比较QAMS法和外标法测定结果的差异。结果在优化色谱条件下,确立的f_(s/i)分别为f_(CYM/CYG)=0.96、f_(FMG/CYG)=0.84、f_(CY/CYG)=0.68、f_(FMM/CYG)=0.86、f_(FM/CYG)=0.53,f_(s/i)在3种HPLC色谱仪/3个品牌C_(18)柱、不同体积流量(0.9、1.0、1.1m L/min)和不同柱温(25、30、35℃)下的RSD分别为0.88%~1.63%、0.23%~0.73%、0.15%~0.46%,表明相对校正因子的耐用性良好。分别采用外标法(ESM)和QAMS法对16批黄芪药材和3批饮片中6个异黄酮及苷类成分进行含量测定,2种方法所得结果的相对误差均<5.0%,无显著性差异,表明建立的QAMS法的准确度良好。结论建立同时测定黄芪中6个异黄酮及苷类化学成分的QAMS方法准确可靠,能为更加全面地评价黄芪药材及饮片的质量提供参考。

Objective To establish a quantitative determination analysis for six isoflavonoid components in Huangqi(Astragali Radix)based on multi-component by a single-marker(QAMS)method.The feasibility and technical adaptability of this approach were investigated in quality evaluation of Astragali Radix.Methods Five isoflavonoids,calycosin-7-O-glycoside-6''-O-malonate(CYM),formononetin-7-O-glycoside(FMG),formononetin-7-O-glycoside-6''-O-malonate(FMM),calycosin(CY)and formononetin(FM)were used as index components to establish the relative correction factor(RCFs)between each component and internal reference substance calycosin-7-O-β-D-glycoside(CYG).The robustness and durability of the measured RCFs of the five components were evaluated on different analytical columns and HPLC instruments.And the measurement result deviation was compared between QAMS method and the external standard method(ESM).Results Under the established chromatographic conditions,the RCFs of f_(CYM/CYG),f_(FMG/CYG),f_(CY/CYG),f_(FMM/CYG),f_(FM/CYG)were determined as 0.96,0.84,0.680.86 and 0.53,respectively.The relative standard deviations(RSD)of their RCFs on three brands of column and HPLC instruments were 0.88%—1.63%,0.23%—0.73%and 0.15%—0.46%,respectively at different flow rates(0.9 mL/min,1.0 mL/min,1.1 mL/min)and different column temperatures(25,30,35℃),indicating the good durability of the RCFs.The present QAMS method and the ESM method were both used to determine the contents of six isoflavonoids from Astragali Radix in 16 batches of crude materials and three batches of decoction pieces.The RSD of content results between two methods were less than 5.0%.It demonstrated that there was no significant difference from content results between these two methods,exhibiting good accuracy of the present QAMS method.Conclusion The present QAMS analytical method for simultaneous determination of six isoflavonoids in Astragali Radix provides a more efficient method to evaluate the comprehensive quality of Astragali Radix.