白花蛇舌草药材的HPLC指纹图谱研究

目的:建立白花蛇舌草药材HPLC指纹图谱,结合化学计量学手段,对多批次药材进行质量控制。方法:采用C18柱(250 mm×4.6 mm,5μm),以乙腈-水(各含0.1‰乙酸)为流动相梯度洗脱,流速0.8 mL/min,检测波长238 nm,柱温30℃;建立白花蛇舌草药材HPLC指纹图谱,通过比对化学分离对照品的保留时间,对主要特征峰进行化学指认;对22批药材进行相似度评价,并进行主成分分析和聚类分析。结果:建立了专属性、精密度、重现性和稳定性均较好的白花蛇舌草药材HPLC指纹图谱;11个特征峰得到明确化学指认;17批白花蛇舌草指纹图谱相似度值明显高于5批伪品;主成分分析结果显示,17批白花蛇舌草密集分布在一个区域,而5批伪品离散分布在此区域外;对22批药材进行聚类分析,17批白花蛇舌草聚为一类,5批伪品均未与其聚为一类;进一步运用聚类分析对17批白花蛇舌草进行质量评价,总共可聚为4类。结论:将HPLC指纹图谱与化学计量学手段相结合,可对白花蛇舌草进行真伪鉴别和质量评价,可为提高其整体质量控制方法提供参考。

Study on HPLC Fingerprint of Oldenlandia diffusa

Objective:To establish the HPLC fingerprint chromatogram of Oldenlandia diffusa coupled with chemometrics means for the quality control of multi-batches of medicinal material.Methods:The separation was developed on C18 column(4.6 mm×250 mm,5 μm) by gradient elution with acetonitrile-water(both containing 0.1‰(V/V)ocetic acid) as mobile phase at a flow rate of 0.8 mL/min,the detection wavelength at 238 nm and column temperature at 30 ℃.The HPLC fingerprint chromatogram of Oldenlandia diffusa was set up and the main characteristic peaks were identified by comparing with chemical reference substance.The quality of 22 batches of medicinal material was evaluated by similarity assay as well as principal component analysis(PCA) and cluster analysis.Results:The established HPLC fingerprint chromatogram of Oldenlandia diffusa was specific,precise,reproducible and stable.11 peaks were chemically identified.The similarity of 17 batches of Oldenlandia diffusa was obviously higher than 5 batches of adulterants.PCA showed that 17 batches of Oldenlandia diffusa were in a domain and 5 batches of adulterants were far apart from the domain.The cluster analysis of the 22 batches of medicinal material showed that 17 batches of Oldenlandia diffusa were in a cluster while 5 batches of adulterants were excluded.Further cluster analysis was carried out for the quality consistency of 17 batches of Oldenlandia diffusa and accordingly they were devided into 4 clusters.Conclusion:With the combination of chemometrics means,the HPLC fingerprint chromatogram provides a method for evaluation of authenticity and quality control of Oldenlandia diffusa,which is favorable to improve overall quality control of Oldenlandia diffusa.