HPLC-一测多评法测定黄精及其饮片中6种成分的含量

Simultaneous Determination of 6 Ingredients in Polygonati Rhizoma and Its Decoction Piece by HPLC-QAMS

目的:建立黄精及其饮片中薯蓣皂苷元、5-羟甲基糠醛、香草酸、芦丁、槲皮素及山柰酚6种成分的一测多评(QAMS)法。方法:采用高效液相色谱(HPLC)外标法同时测定黄精及其饮片中6种成分的含量[色谱柱为Diamonsil-C18;流动相为乙腈-水(梯度洗脱);5-羟甲基糠醛、香草酸、芦丁、槲皮素及山柰酚检测波长为254nm(0~60min),薯蓣皂苷元检测波长为202nm(60~75min);流速为1.0mL/min;柱温为30℃]。以香草酸为内标,分别计算出其余5种成分的相对校正因子(RCFs)并进行耐用性考察,并采用相对保留法对待测成分色谱峰进行准确定位,然后根据RCFs计算黄精药材中其他5种成分的含量,并与外标法测定结果进行比较。以黄精饮片进行方法验证,分别采用QAMS法和外标法测定其中6种成分的含量,比较两种方法含量测定差异。结果:HPLC方法学考察结果均符合相关要求。在线性范围内,薯蓣皂苷元、5-羟甲基糠醛、芦丁、槲皮素及山柰酚的RCFs分别为0.195、0.025、0.263、0.345、0.075,在不同试验条件下RCFs重现性良好。外标法和QAMS法两种方法测得的黄精药材及其饮片中6种成分的含量差异均无统计学意义(P>0.05)。结论:建立的QAMS法可用于同时测定黄精药材及其饮片中6种成分含量。

OBJECTIVE:To establish a quantitative analysis of multi-components by singer marker(QAMS)for determining the contents of diosgenin,5-hydroxymethylfurfural,vanillic acid,rutin,quercetin,kaempferol in Polygonati Rhizoma and its decoction piece.METHODS:HPLC external standard method was used to determine the contents of 6 components in Polygonati Rhizoma and its decoction piece simultaneously[the separation was carried out on Diamonsil-C18 column with mobile phase consisted of acetonitrile-water(gradient elution);the detection wavelengths of 5-hydroxymethylfurfural,vanillic acid,rutin,quercetin and kaempferol were set at 254 nm(0-60 min);the detection wavelength of diosgenin was set at 202 nm(60-75 min)at the flow rate of 1.0 mL/min.The column temperature was 30℃].Using vanillic acid as internal standard,relative correction factors(RCFs)of aother 5 components were calculated and to investigate durability.Relative retention method was used to accurately locate the chromatographic peaks of the components to be determined,and then the contents of the aother 5 components in Polygonati Rhizoma were calculated according to RCFs,and the results were compared with those determined by external standard method.The method was validated by Polygonati Rhizoma decoction piece.The contents of 6 components were determined by QAMS method and external standard method respectively,and then the differences of content determination were compared between 2 methods.RESULTS:The methodology investigation results of HPLC method were in line with related requirements.Within the linear range,the RCFs of diosgenin,5-hydroxymethylfurfural,rutin,quercetin,kaempferol were 0.195,0.025,0.263,0.345 and 0.075,respectively.Under different experiment conditions,RCFs showed good reproducibility;there was no statistical significance of 6 components in Polygonati Rhizoma and its decoction piece determined by external standard method and QAMS method(P>0.05).CONCLUSIONS:Established QAMS method is suitable for simultaneous determination of 6 components in Polygonati Rhizoma and its decoction piece.