一测多评法测定止血化痔合剂中6种黄酮类成分的含量

Determination of six flavonoids in Zhixue Huazhi mixture by quantitative analysis of multicomponents

目的建立一种一测多评法(QAMS)同时测定止血化痔合剂中6种黄酮类成分的含量。方法采用高效液相色谱法,以黄芩苷为内参物,建立黄芩苷与芦丁、甘草苷、槲皮素、黄芩素、汉黄芩素的相对校正因子(f),再通过f计算出止血化痔合剂中6种黄酮类成分的含量。同时采用外标法(ESM)测定6种黄酮类成分的含量,并比较二者的差异,以验证一测多评法的准确性和可行性。结果芦丁、甘草苷、黄芩苷、槲皮素、黄芩素、汉黄芩素在一定范围内浓度与峰面积呈良好的线性关系(r>0.997);精密度、稳定性、重复性均符合方法学要求;平均加样回收率范围为95.72%~100.43%,RSD范围为1.97%~3.22%。芦丁、甘草苷、槲皮素、黄芩素、汉黄芩素的相对校正因子分别为6.426、3.939、3.215、41.140、0.555,且重现性良好。一测多评法的计算结果与外标法测定结果差异无统计学意义(P>0.05)。结论以黄芩苷为内参物建立的一测多评法,同时测定止血化痔合剂中6种黄酮类成分的含量,方法操作简便,准确可靠,可用于止血化痔合剂的质量评价。

Objective To develop a method for the simultaneous determining the content of six flavonoids in Zhixue Huazhi mixture by a quantitative analysis of multi-components with single marker(QAMS).Methods Baicalin was taken as internal standard substance to establish the relative correction factor(f)of rutin,liquiritin,quercetin,baicalein,wogonin by HPLC.The contents of above six flavonoids were calculated through f.Meanwhile,external standard method(ESM)was used to calculate the content of six flavonoids.The difference of two methods were compared to validate the accuracy and feasibility of QAMS.Results The standard curves of rutin,liquiritin,baicalin,quercetin,baicalein,wogonin had good linear relationship in ranges of the tested concentrations(r>0.997).The precision,stability and repeatability complied with the requirements of methodology.The average recoveries were 95.72%-100.43%,and the RSD were 1.97%-3.22%.The relative correction factors of rutin,liquiritin,quercetin,baicalein,and wogonin were 6.426,3.939,3.215,41.140,0.555,with good repeatability.There was no significant difference of the quantitative results between QAMS and ESM method(P>0.05).Conclusions The QAMS established with baicalin as internal standard substance simultaneously determine the content of six flavonoids in Zhixue Huazhi mixture.The method is simple,accurate and reliable,and can be used for the quality evaluation of Zhixue Huazhi mixture.