基于一测多评定量控制联合灰色关联度分析的颈痛颗粒综合质量评价

Comprehensive quality evaluation of Jingtong Granules based on HPLC-QAMS multi-index component quantitative control combined with grey relational analysis

目的建立高效液相色谱一测多评(QAMS)法同时测定颈痛颗粒中三七皂苷R_(1)、人参皂苷Rg_(1)、人参皂苷Rb1、3'-羟基葛根素、葛根素、3'-甲氧基葛根素、洋川芎内酯A、藁本内酯、延胡索乙素、去氢紫堇碱、紫堇碱、羌活醇和异欧前胡素含量,并通过灰色关联度分析法对13种成分含量检测结果进行分析评价。方法采用Shimadzu C_(18)色谱柱;以乙腈-0.1%磷酸为流动相梯度洗脱;检测波长为203 nm(三七皂苷R_(1)、人参皂苷Rg_(1)和人参皂苷Rb1)、280 nm(3'-羟基葛根素、葛根素、3'-甲氧基葛根素、洋川芎内酯A、藁本内酯、延胡索乙素、去氢紫堇碱和紫堇碱)和315 nm(羌活醇和异欧前胡素);采用外标法(ESM)测定13种成分的含量。以葛根素为内参物质,分别计算其与三七皂苷R_(1)、人参皂苷Rg_(1)、人参皂苷Rb1、3'-羟基葛根素、3'-甲氧基葛根素、洋川芎内酯A、藁本内酯、延胡索乙素、去氢紫堇碱、紫堇碱、羌活醇和异欧前胡素的相对校正因子(f),并计算上述12种成分的含量,比较ESM和QAMS法检测结果的差异,验证一测多评法的准确性和可行性。基于灰色关联度分析不同批次颈痛颗粒中13种成分QAMS法含量检测数据,对15批次颈痛颗粒质量进行综合评价。结果三七皂苷R_(1)、人参皂苷Rg_(1)、人参皂苷Rb1、3'-羟基葛根素、葛根素、3'-甲氧基葛根素、洋川芎内酯A、藁本内酯、延胡索乙素、去氢紫堇碱、紫堇碱、羌活醇和异欧前胡素分别在各自范围内线性关系良好(r≥0.9992),平均加样回收率(n=9)在96.86%~100.37%(RSD≤1.65%)。所建立的f耐用性良好,HPLC-QAMS法计算值和ESM实测值之间无明显差异。灰色关联度分析结果显示15批次颈痛颗粒相对关联度在0.3324~0.6004,表明颈痛颗粒批次间呈现一定质量差异。结论QAMS多指标成分定量控制联合灰色关联度分析法操作便捷、结果准确,可用于颈痛颗粒的综合质量评价。

Objective To establish a HPLC-QAMS method for simultaneous determination of notoginsenoside R_(1),ginsenoside Rg_(1),ginsenoside Rb1,3'-hydroxy puerarin,puerarin,3'-methoxy puerarin,senkyunolide A,ligustilide,tetrahydropalmatine,dehydrocorydaline,corydaline,notopterol and isoimperatorin in Jingtong Granules,and to analyze and evaluate the content detecting results of 13 components by grey relational analysis.Methods The chromatographic column Shimadzu C_(18) was used.And using acetonitrile-0.1%phosphoric acid as mobile phase with gradient elution.The detection wavelength were 203 nm for notoginsenoside R_(1),ginsenoside Rg_(1),ginsenoside Rb1,280 nm for 3'-hydroxy puerarin,puerarin,3'-methoxy puerarin,senkyunolide A,ligustilide,tetrahydropalmatine,dehydrocorydaline,corydaline and 315 nm for notopterol and isoimperatorin.The contents of 13 components were determined by external standard method(ESM).Taking puerarin as internal standard substance,the relative correlation factor(f)of notoginsenoside R_(1),ginsenoside Rg_(1),ginsenoside Rb1,3'-hydroxy puerarin,3'-methoxy puerarin,senkyunolide A,ligustilide,tetrahydropalmatine,dehydrocorydaline,corydaline,notopterol and isoimperatorin was calculated,and the contents of 12 components were calculated.The differences between the content detecting results by ESM and HPLC-QAMS method were compared.Based on the grey relational degree,the HPLC-QAMS method conent detection data of 13 component of different batches of Jingtong Granules were analyzed,and the sample of 15 batches of Jingtong Granules was comprehensively quality evaluated.Results Notoginsenoside R_(1),ginsenoside Rg_(1),ginsenoside Rb1,3'-hydroxy puerarin,puerarin,3'-methoxy puerarin,senkyunolide A,ligustilide,tetrahydropalmatine,dehydrocorydaline,corydaline,notopterol and isoimperatorin had good linearity in their respective ranges(r≥0.9992).The average recoveries of the 13 components(n=9)were between 96.86%-100.37%with the relative standard deviations less than 1.65%.The established RCF had good durability,and the results calculated with HPLC-QAMS method were consistent with the results by the ESM.The results of grey correlation analysis showed that the relative correlation degree of 15 batches of Jingtong Granules was 0.3324-0.6004,indicating that there was a certain quality difference between batches of Jingtong Granules.Conclusion HPLC-QAMS multi-index component quantitative control combined with grey relational analysis is convenient and accurate,and can be used for the comprehensive quality evaluation of Jingtong Granules.