基于指纹图谱结合多成分定量分析的视疲宁片质量评价研究

目的:建立视疲宁片(SPN)的HPLC特征指纹图谱,结合聚类分析、主成分分析、多成分定量分析,为其质量评价提供依据。方法:采用Agilent C18柱(250mm×4.60mm,5μm),以流动相为乙腈-0.1%磷酸水溶液,梯度洗脱(0～15min,20%～25%乙腈,20～25min,25%～45%乙腈,25～30min,45%～95%乙腈);体积流量为1mL·min-1;检测波长为280nm;柱温30℃。采用《中药色谱指纹图谱相似度评价系统》(2012年版)对10批SPN指纹图谱进行相似度评价,对共有峰进行聚类分析与主成分析,并对指认的6种指标成分进行定量分析。结果:10批SPN HPLC指纹图谱相似度均大于0.90,共20个共有峰,各峰分离度较好。10批SPN样品聚类分析与主成分分析结果具有一致性,且经主成分分析20个共有峰可综合分为6个主成分,累积方差贡献率为95.91%;通过与对照品比对确定6种指标成分,分别为绿原酸(3号峰)、阿魏酸(5号峰)、橙皮苷(7号峰)、淫羊藿苷(12号峰)、橙黄决明素(18号峰)、大黄酚(20号峰),所建立的成分分析方法可用于6种成分的含量测定。结论:HPLC特征指纹图谱结合模式识别可反映SPN的内在质量,建立的多成分定量分析方法对SPN的质量控制具有参考价值。

Study on quality evaluation of Shipining tablets by HPLC fingerprint and multi-component quantitation

OBJECTIVE To establish the HPLC chemical fingerprints of Shipining tablets and make the multicomponent quantitative analysis to provide the basis for its quality evaluations by combining with cluster analysis and principal component analysis (PCA). METHODS The analysis was carried out on an analytical column Agilent C₁₈ (250 mm×4.6 mm, 5 μm) with gradient elution by methyl cyanides (A)-0.1% phosphoric acid solution (B) (0-15 min, 20%-25% A; 20-25 min, 25%-45% A; 25-30 min, 45%-95% A), at the detection wavelength of 280 nm and a flow rate of 1.0 mL/min. The column temperature was 30℃. Additionally, fingerprint similarity of 10 batches of Shipining tablets was evaluated by software "Similarity Evaluation System for Chromatographic Fingerprint of TCM" issued by GPC (Version 2012). PCA was used for common peaks. The amounts of six characteristic components in Shipining tablets were also simultaneously determined. RESULTS The common fingerprint pattern derived from 10 batches of samples was obtained with the similarity over 0.90 and 20 common peaks were defined. Ten Batches SPN's cluster analysis results and PCA results were coincident. According to PCA, the 20 peaks could be integrated into 6 principal components, with cumulative contribution rate of principal component of 95.91%. Meanwhile, some common peaks were identified via peak pattern matched with standard substances, showing that the peak No.3, No.5, No.7, No.12, No.18 and No.20 were chlorogenic acid, ferulic acid, hesperidin, icariin, aurantio-obtusinnd and chrysophanol. The composition analysis method established could be used for the determination of the contents of 6 components. CONCLUSION HPLC chemical fingerprints combined with pattern recognition can reflect the intrinsic quality of SPN. And the multi component quantitative analysis method is of reference value for the quality control of SPN.