紫花地丁高效液相色谱法指纹图谱的建立和评价

摘要

用高效液相色谱法（HPLC）建立紫花地丁的指纹图谱,对来源于6个产地的10批紫花地丁药材进行分析。采用VP-ODS色谱柱为固定相,由不同比例的甲醇和磷酸（0.4＋99.6）溶液配成的混合溶液作为流动相进行梯度淋洗,对不同时段的洗脱液在波长350nm处作紫外检测,建立了10批药材的色谱指纹图谱的共有模式。用紫外分光光度法、红外光谱法、质谱法及核磁共振波谱法对柱色谱法分离所得紫药地丁的主要色谱峰代表的成分进行了分析和鉴定,确证了该组分的结构为秦皮乙素。以秦皮乙素峰为参照峰,对10批药材指纹图谱进行相似度分析和聚类分析,10批紫花地丁药材均能被明显地区分为二类。%HPLC was applied to the establishment of chromatographic fingerprints of Viola yedoensis.The HPLC fingerprints of ten samples of Viola yedoensis from six growing areas were determined.VP-ODS chromatographic column was used as stationary phase,and mixtures of methanol and H3PO4（0.4＋99.6） solution with various proportions were used as mobile phase in the gradient elution,eluates obtained at various time intervals were detected at the wavelength of 350 nm.Common pattern of HPLC finger prints of the 10 samples was established.Components referring to the main chromatographic peaks in the chromatograms obtained by HPLC separation of the Viola yedoensis samples were analyzed and identified further by UV-spectrophotometry,IR-spectrometry,MS and NMR spectrometry,and were ascertained to be the same compound of aesculetin.Using the chromatographic peak of aesculetin as a reference peak,the fingrprints of the 10 samples of Viola yedoensis were studied by the similarity analysis and clustering analysis showing that the 10 batches of Viola yedoensis from 6 origins were classified into 2 categories.