Establishment and Evaluation of HPLC Fingerprints of Viola Yedoensis Makino

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.