Comparative analysis of major constituents in Viola yedoensis Makino and different species from the Genus Viola by high-performance liquid chromatography with chemometrics methods

Abstract

The whole plant of Viola yedoensis Makino is used for the treatment of rhinitis, rheumatism, mastitis and acute pyogenic infections. A simple and accurate HPLC-UV method was developed for the simultaneous determination of six major constituents in the whole plant ofV. yedoensis, and the comparison between its constituent profile and eight other species in the genus Viola was investigated. The optimal chromatographic conditions were achieved on a Hedera ODS-2 (250 mm×4.6 mm, i.d. 5 μm) column with gradient elution using 0.4% acetic acid aqueous and methanol at 354 nm. Calibration curves presented good linear regression (r > 0.9996) within test ranges. The overall recoveries ranged between 96.72 and 97.67% and the RSDs were less than 2.34%. The results indicated that esculetin was the major components in V. yedoensis. Principal component analysis and cluster analysis could evaluate the quality and distinguish V. yedoensis from eight other species in the genus Viola. The HPLC technique combined with chemometrics was a sensitive, selective, rapid and guiding method and could be applied to the further quality evaluation of complex materials. Key words: Viola yedoensis, coumarin, simultaneous determination, Viola, chemometrics method.