紫花地丁HPLC特征图谱结合多组分含量测定方法研究

Study on HPLC characteristic chromatogram of Violae Herba combined with multi component content determination

目的:建立紫花地丁的HPLC特征图谱,并测定秦皮甲素、秦皮乙素的含量。方法:色谱柱为CAPCELL PAK C18色谱柱(4.6mm×250mm,5μm);流动相为甲醇(A)-0.4%磷酸(B)梯度洗脱,流速为1.0mL/min;检测波长为327nm;柱温为35℃,建立紫花地丁HPLC特征图谱。运用"中药色谱指纹图谱相似度评价软件(2012版)"和SPSS 20.0软件进行相似度分析、主成分分析和聚类分析。结果:建立了紫花地丁药材的特征图谱,确立了9个特征峰,19批紫花地丁的相似度为0.814~0.996;秦皮甲素、秦皮乙素含量分别为0.51~4.32、1.95%~11.76%;聚类分析将紫花地丁分为3类,S9样品单独聚为一类,可能与其生长环境有关。结论:建立了紫花地丁的HPLC特征图谱结合含量测定的方法,为其质量控制和品质评价提供参考。

Objective: The HPLC characteristic spectrum of Violae Herba was established, and the contents of esculine and aesculetin were determined. Methods: The chromatographic column was CAPCELL PAK C18(4.6 mm×250 mm, 5μm). The mobile phase was methanol(A)-0.4% phosphoric acid(B) gradient elution, and the flow rate was 1.0 mL/min. The detection wavelength was 327 nm;The column temperature was 35℃. Similarity analysis, principal component analysis and cluster analysis were carried out by using the similarity evaluation software of TCM fingerprint(2012 edition) and SPSS 20.0 software. Results: The characteristic spectrum of Violae Herba was established, and 9 characteristic peaks were established. The similarity of 19 batches of Violae Herba was 0.814~0.996. The contents of esculine and aesculetin were 0.51~4.32, 1.95%~11.76%;According to the cluster analysis, the S9 samples was grouped into three categories, which may be related to their growth environment. Conclusion: A HPLC method was established for the determination of the content of Violae Herba, which provided a reference for quality control and quality evaluation.