葛根和粉葛高效液相色谱指纹图谱及化学模式识别

HPLC Fingerprint and Chemical Pattern Recognition of Pueraria lobata and Pueraria thomsonii

目的:建立并比较葛根和粉葛的高效液相色谱(HPLC)指纹图谱,为其质量控制提供依据。方法:采用HPLC法,色谱柱为Kromasil C_(18)(4.6 mm×200 mm),流动相为乙腈-0.03 mol/L醋酸铵溶液(梯度洗脱),流速为0.9m L/min,柱温为40℃,检测波长为250 nm,进样量为20μL。收集16批次的葛根和粉葛样品进行测定,采用《中药色谱指纹图谱相似度评价系统2004A版》软件进行相似度评价,采用聚类分析和主成分分析对指纹图谱进行化学模式识别研究。结果:16批葛根和粉葛药材的指纹图谱有6个共有峰,经对照指认出葛根素、大豆苷、大豆苷元和染料木素的色谱峰。相似度分析结果显示葛根和粉葛的相似度较高,均在0.910以上;聚类分析将葛根和粉葛样品分别归为两类;主成分分析提取得到8个主成分,累计方差贡献率达到93.548%,前3个主成分得分投影结果与聚类分析基本一致。结论:葛根和粉葛所含化学成分既有一定的相似性,也存在一定差异,该方法可作为葛根和粉葛的质量控制及临床应用的依据。

Objective： To establish and compare HPLC fingerprints of Pueraria lobata and Pueraria thomsonii. Methods： HPLC was used to analyze16 batches of samples. The determination was performed on Kromasil C_（18） column with mobile phase consisted of acetonitrile- 0. 03 mol/L ammonium acetate with gradient elution at the flow rate of 0. 9 m L/min. The detection wavelength was 250 nm and column temperature was set at 40 ℃. The sample size was 20 μL. TCM fingerprint evaluation software（ 2004 edition） was used to evaluate the similarity of 16 batches of samples. Hierarchical cluster analysis（ HCA） and principal component analysis（ PCA） were applied to chemical pattern recognition. Results： There were 6 common peaks of fingerprints of Pueraria lobata and Pueraria thomsonii. Four components were identified as Puerarin,Daidzin,Daidzein and Genistein. The similarities of 16 batches of samples were proved to be higher than 0. 910. Sixteen batches of Pueraria lobata sample and Pueraria thomsonii ware classified into two groups（ Pueraria lobata sample and Pueraria thomsonii sample） based on HCA. Eight principal components（ PC） were observed via PCA and the values of 8 principal components accounted for 93. 548% of the total variance. The classification result according to PCA was consistent with the HCA. Conclusion： The components of Pueraria lobata and Pueraria thomsonii are different. The method can be used for identification and quality evaluation of Pueraria lobata and Pueraria thomsonii.