基于聚类和主成分分析的红花抗肝纤维化提取物特征图谱研究

Study on specific chromatogram of anti-hepatic fibrosis extract from Carthamus tinctorius L. coupled with cluster analysis and principal component analysis

目的利用高效液相色谱技术和数理统计分析方法建立一种红花药材的鉴别和质量控制方法。方法采用高效液相色谱法,用安捷伦C18(4.6 mm×250 mm,5μm)色谱柱,以乙腈-水为梯度洗脱相;流速为1 m L/min,柱温为30℃,检测波长为280 nm。检测了16批不同来源的红花样品具有抗肝纤维化作用的提取物,通过相似度分析建立了红花提取物的高效液相色谱特征图谱。对不同产地的红花提取物特征图谱进行聚类分析和主成分分析,并用《中药色谱指纹图谱相似度评价系统(2004A版)》对其质量进行评价。结果本研究建立了16批红花药材提取物的指纹图谱共有模式,确定了32个共有色谱峰。结论本方法稳定性、重现性好,所建立的红花抗肝纤维化提取物的特征图谱特征性和专属性强,可以用于高质量红花药材的鉴别和质量控制。

Objective To establish and validate a high performance liquid chromatography （HPLC） coupled with mathematical statistics analysis for identification and quality control of different kinds of Carthamus tinctorius L.. Methods The high performance liquid chromatography analysis was used to determine 16 batches of anti-hepatic fibrosis extract samples from Carthamus tinc torius L. on an Agilent C18 column （4.6 mm＆#237;250 mm, 5μm） with the gradient elution solvent system composed of acetonitrile （A） and water （B）. The flow rate was 1 mL/min, the column temperature was maln-talned at 30℃ and the wavelength was acquired at 280 nm. Similarity analysis was used to establish the reference fingerprint spectra. The extract HPLC chromatograms from Carthamus tinc torius L. with anti-hepatic fibrosis activity were analyzed by hierarchical clustering analysis （HCA） and principle component analysis （PCA）. The chromatographic fingerprint of traditional Chinese medicine （version： 2004A） was assessed by similarity evaluation system to evaluate the quality. Results The 16 batches of Carthamus tinc torius L. fingerprints were established. The 32 peaks were determined as the common peaks. Conclusion From the results, we know that it is a stable and reproducible method, which can be used for identification and quality control of Carthamus tinc torius L. with the specific fingerprint.