HPLC多指标成分联合PCA和OPLS-DA法的复方儿茶胶囊综合质量评价

Comprehensive quality evaluation of Compound Ercha Capsules by HPLC with PCA and OPLS-DA method

目的建立HPLC法同时检测复方儿茶胶囊中儿茶素、表儿茶素、槲皮素和山奈酚的含量,并结合主成分分析(PCA)、正交偏最小二乘法-判别分析(OPLS-DA)法构建其质量控制体系。方法采用Waters Symmetry C18色谱柱(5μm,250 mm×4.6 mm),流动相乙腈-0.5%冰醋酸,梯度洗脱;检测波长分别为280 nm(检测儿茶素和表儿茶素)、360 nm(检测槲皮素和山奈酚);用外标法计算各成分含量,并联合PCA、OPLS-DA进行品质分析。结果4种成分分别在各自范围内线性关系良好(r≥0.9992);平均加样回收率96.85%~99.54%(RSD<2.0%);经PCA法得2个主成分的累积贡献率达到87.463%;OPLS-DA法显示12批复方儿茶胶囊样品聚为3类;儿茶素和表儿茶素是影响复方儿茶胶囊产品质量的主要潜在标志物。结论所建方法结果准确、操作便捷,可用于复方儿茶胶囊的综合质量评价。

Objective To establish an HPLC method for simultaneous determination of catechin,epicatechin,quercetin and kaempferol in Compound Ercha Capsules,and to construct its quality control system by combining principal component analysis(PCA)and orthogonal partial least squares-discriminant analysis(OPLS-DA).Methods The separation was carried out on Waters Symmetry C18 column(5μm,250 mm×4.6 mm).The mobile phase consisted of acetonitrile-0.5%glacial acetic acid with gradient elution.The detection wavelengths were 280 nm for catechin and epicatechin,360 nm for quercetin and kaempferol.The content of each component was calculated by external standard method,and the quality was analyzed by PCA and OPLS-DA.Results The 4 components had good linear relationships within their respective ranges(r≥0.9992).The average recovery were 96.85%~99.54%(RSD<2.0%).The cumulative contribution rate of the two principal components obtained by PCA method reached 87.463%.The OPLS-DA method showed that 12 batches of compound Ercha capsules were clustered into three categories,catechin and epicatechin were the main potential markers affecting the quality of Compound Ercha Capsules.Conclusion The method is accurate and easy to operate,and can be used for the comprehensive quality evaluation of Compound Ercha Capsules.