基于一测多评法测定银黄制剂中6种咖啡酰奎宁酸类成分的含量

Simultaneous determination of six caffeoylquinic acids on Yinhuang preparations by quantitative analysis of multi-components by single marker

[目的]建立测定银黄制剂中6种咖啡酰奎宁酸类成分(绿原酸、新绿原酸、隐绿原酸、异绿原酸A、异绿原酸B和异绿原酸C)含量的一测多评(QAMS)方法.[方法]采用Diamonsil C_(18)(4.6 mm×250.0 mm,5.0μm)色谱柱,以0.4%(体积分数)磷酸水溶液(A)-乙腈(B)为流动相进行梯度洗脱(流动速度为1.0 m L·min^(-1),进样量10μL,检测波长327 nm,柱温30℃).以绿原酸(5-O-咖啡酰奎宁酸)为内参物,计算其他5种咖啡酰奎宁酸类成分的相对校正因子并计算该5种成分的含量,实现QAMS.同时采用外标法测定6种咖啡酰奎宁酸类成分的含量,并对两种方法测定的结果进行比较,验证QAMS法的可行性和准确性.[结果]在一定线性范围内,以绿原酸为内参,新绿原酸(3-O-咖啡酰奎宁酸)、隐绿原酸(4-O-咖啡酰奎宁酸)、异绿原酸C(4,5-O-二咖啡酰奎宁酸)、异绿原酸A(3,5-O-二咖啡酰奎宁酸)和异绿原酸B(3,4-O-二咖啡酰奎宁酸)5种成分的平均校正因子分别为1.2243、0.9569、0.8141、0.8838和0.7599.QAMS与外标法测定的结果间无明显差异.[结论]成功建立了同时测定银黄系列制剂中6种咖啡酰奎宁酸的QAMS法,该方法简便、快速、准确、可行,可用于银黄制剂产品多指标成分的质量控制与评价.

OBJECTIVE To establish a quantitative analysis of multi-components by single marker(QAMS)method for the determination of six caffeoylquinic acids(5-O-caffeoylquinic acid,3-O-caffeoylquinic acid,4-O-caffeoylquinic acid,4,5-O-dicaffeoylquinic acid,3,5-O-dicaffeoylquinic acid and 3,4-O-dicaffeoylquinic acid)in Yinhuang preparations.METHODS Diamonsil C_(18)column(4.6 mm×250.0 mm,5.0μm)was used for gradient elution using 0.4%(volume fraction)phosphoric acid aqueous solution(A)-acetonitrile(B)as mobile phase(flow rate was 1.0 mL·min^(-1),sample size was 10μL,detection wavelength was 327 nm,and the column temperature was 30℃).5-O-caffeoylquinic acid was selected as the internal reference,the relative correction factors(RCFs)of the other five caffeoylquinic acids were calculated,and the contents of the five constituents were calculated to realize QAMS.At the same time,the external standard method was used to determine the contents of the six caffeoylquinic acids constituents.The results of the two methods were compared to verify the feasibility and accuracy of QAMS method.RESULTS Within a certain linear range,5-O-caffeoylquinic acid as the internal reference,the average RCFs of 3-O-caffeoylquinic acid,4-O-caffeoylquinic acid,4,5-O-dicaffeoylquinic acid,3,5-O-dicaffeoylquinic acid and 3,4-O-dicaffeoylquinic acid were 1.2243,0.9569,0.8141,0.8838 and 0.7599 respectively.There was no significant difference between QAMS and external standard method.CONCLUSION The QAMS method for the simultaneous determination of six caffeoylquinic acids in Yinhuang preparations was established successfully.This method is simple,rapid,accurate and feasible,and can be used for the quality control and evaluation of multiple index components in Yinhuang preparations.