基于高效液相色谱-二极管阵列检测器法的复康宁胶囊有效成分含量测定的方法学研究

Methodological Study on the Assay of Effective Components in Fukangning Capsules Based on HPLC-DAD

目的:基于高效液相色谱-二极管阵列检测器法(high performance liquid chromatography-diode array detection,HPLC-DAD)构建复康宁胶囊有效成分的含量测定方法,为复康宁胶囊的质量评价提供依据。方法:采用HPLC-DAD,以Agilent C18柱(250 mm×4.6 mm,5μm)作为色谱柱,动相为乙腈(A)-0.1%磷酸溶液(B),切换检测波长进行梯度洗脱,流速为0.8 mL/min,柱温为30℃;测定复康宁胶囊中野黄芩苷、野黄芩素、太子参环肽B、车叶草苷酸和车叶草苷5种有效成分的含量。结果:分析结果显示,野黄芩苷、野黄芩素、太子参环肽B、车叶草苷酸和车叶草苷的回归方程分别为y=6.9835×10^(5)x+268.7(R=0.9999)、y=6.1137×10^(5)x+412.6(R=0.9998)、y=5.3456×10^(5)x-318.9(R=0.9997)、y=8.9742×10^(5)x-374.4(R=0.9996)和y=1.1028×10^(6)x-439.8(R=0.9997),并且5种有效成分的专属性、精密度、稳定性、重复性和回收率试验均非常良好;含量测定结果显示,复康宁胶囊中野黄芩苷、野黄芩素、太子参环肽B、车叶草苷酸和车叶草苷的含量分别为19.878~20.371 mg/g、10.788~11.122 mg/g、1.782~2.013 mg/g、0.762~0.812 mg/g和1.087~1.234 mg/g。结论:采用HPLC-DAD测定复康宁胶囊中野黄芩苷、野黄芩素、太子参环肽B、车叶草苷酸和车叶草苷具有良好的专属性、精密度、稳定性、重复性和回收率,并且测定方法快速、简便、准确,是一种较好的复康宁胶囊质量控制方法。

Objective:To establish a method for the assay of effective components in Fukangning Capsules based on high performance liquid chromatography-diode array detection(HPLC-DAD),and provide a basis for the quality evaluation of Fukangning Capsules.Methods:The HPLC-DAD was adopted,Agilent C18 column(250 mm×4.6 mm,5μm)was used as a chromatographic column,the mobile phase was acetonitrile(A)-0.1%phosphoric acid solution(B),the detection wavelength was switched for gradient elution,the flow rate was 0.8 mL/min,and the column temperature was 30℃.The contents of five effective components in Fukangning Capsules,including baicalin,scutellarin,heterophyllin B,desacetyl asperuloside acid and asperuloside,were determined.Results:The analysis results showed that the regression equations for baicalin,scutellarin,heterophyllin B,desacetyl asperuloside acid and asperuloside were y=6.9835×10^(5)x+268.7(R=0.9999),y=6.1137×10^(5)x+412.6(R=0.9998),y=5.3456×10^(5)x-318.9(R=0.9997),y=8.9742×10^(5)x-374.4(R=0.9996)and y=1.1028×10^(6)x-439.8(R=0.9997),respectively,and the specificity,precision,stability,repeatability and recovery rate of the five effective components were all very good.The assay results showed that the contents of baicalin,scutellarin,heterophyllin B,desacetyl asperuloside acid and asperuloside in Fukangning Capsules were 19.878-20.371 mg/g,10.788-11.122 mg/g,1.782-2.013 mg/g,0.762-0.812 mg/g and 1.087-1.234 mg/g,respectively.Conclusion:HPLC-DAD for the assay of baicalin,scutellarin,heterophyllin B,desacetyl asperuloside acid and asperuloside in Fukangning Capsules has good specificity,precision,stability,repeatability and recovery rate.This assay method is fast,simple and accurate,and is a good quality control method for Fukangning Capsules.