The development and evaluation of an HPLC-DAD method for the analysis of the phenolic fractions from in vivo and in vitro biomass of Hypericum species

A new HPLC-DAD method for separation and identification of the main phenoli cs present in in vivo and in vitro biomass of Hypericum perforatum and Hype ricum androsaemum has been developed. This method accomplished the direct i dentification of 22 compounds including flavonoids, hypericins, phlorogluci nols, and phenolic acids. The HPLC profiles obtained in the course of this work clearly evidenced a distinct phenolic production between in vivo and i n vitro biomass. For example calli and suspended cells produced mainly xant hones while in in vivo plants of both species these compounds were not dete cted. Some luteolin flavone type compounds were identified in calli and suspended cells of H. perforatum while quercetin was found in suspended cells of H. androsaemum. Flavonoids, namely those related to quercetin, were the major metabolites in methanolic extracts from in vivo plants. Hypericins were det ected in in vivo plants and in in vitro shoots of H. perforatum but not in calli or suspended cells of the same species. Aurones, produced by in vitro cultures were not detected in in vivo plants of H. perforatum.