不同产地夏枯草HPLC指纹图谱及化学模式识别研究

Study on HPLC Fingerprint and Chemical Pattern Recognition of Xiakucao(Prunella vulgaris)from Different Producing Areas

目的建立不同产地16批夏枯草HPLC指纹图谱结合化学模式识别技术对夏枯草进行质量评价。方法Agilent Zorbax C_(18)键合硅胶柱(250 mm×4.6 mm,5μm);乙腈-0.1%磷酸水溶液,梯度洗脱;检测波长:210 nm;流速:1 mL/min;进样量:10μL;温度:30℃;建立夏枯草HPLC指纹图谱,采用相似度评价、聚类分析、主成分分析、聚类热图和正交偏最小二乘判别分析等化学模式识别方法对不同产地夏枯草质量及其控制方法进行分析和评价。结果16批夏枯草的定量指纹图谱相似度均>0.88,标定了13个共有峰,指认出咖啡酸、芦丁、金丝桃苷、迷迭香酸、熊果酸等5个共有峰。16批夏枯草可聚为4类,S2、S3、S4、S6、S8、S5、S7、S13聚合为一类,S10、S11、S14聚为一类,S12、S15、S16聚合为一类,S1、S9聚为一类。主成分分析得到3种主成分因子的累计方差贡献率为83.896%,能有效区分不同产地夏枯草,并确定迷迭香酸等3种成分为影响其质量的差异性质量标志物。结论建立的HPLC指纹图谱结合化学模式识别的方法稳定、可靠、简便准确,可以更系统、全面地为夏枯草的质量控制评价提供依据。

Objective To establish the HPLC fingerprints of 16 batches of Xiakucao(Prunella vulgaris)from different producing areas and evaluate the quality of Xiakucao(Prunella vulgaris)by chemical pattern recognition technology.Methods Agilent Zorbax C_(18) bonded silica column(250 mm×4.6 mm,5μm)was used.The conditions included acetonitrile-0.1%phosphoric acid aqueous solution,gradient elution.The detection wavelength was 210 nm and the flow rate was 1 mL/min.The injection volume was 10μL and the temperature was 30℃.The HPLC fingerprint of Xiakucao(Prunella vulgaris)was established by using similarity evaluation,cluster analysis,principal component analysis,clustering heat map and orthogonal partial least squares analysis and other chemical pattern recognition methods to evaluate the quality and its control method was analyzed and evaluated.Results The similarity of quantitative fingerprints of 16 batches of Xiakucao(Prunella vulgaris)was greater than 0.88,13 common peaks were calibrated,and 5 common peaks of caffeic acid,rutin,hyperoside,rosmarinic acid and ursolic acid were identified.16 batches of Prunella vulgaris can be aggregated into 4 categories.S2,S3,S4,S6,S8,S5,S7 and S13 aggregated into one category.S10,S11 and S14 aggregated into one category,and S12,S15 and S16 aggregated into one category,S1 and S9 were clustered into one category.The principal component analysis showed that the cumulative variance contribution rate of the three principal component factors was 83.896%,which could effectively distinguish Xiakucao(Prunella vulgaris)from different producing areas,and identified three components such as rosmarinic acid as differential quality markers affecting its quality.Conclusion The method of HPLC fingerprint and chemical pattern recognition established in this study is stable,reliable,simple and accurate,and can provide a basis for the quality control evaluation of Xiakucao(Prunella vulgaris)more systematically and comprehensively.