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A new HPLC-DAD method for separation and identification of the main phenolics present in in vivo and in vitro biomass of Hypericum perforatum and Hypericum androsaemum has been developed. This method accomplished the direct identification of 22 compounds including flavonoids, hypericins, phloroglucinols, and phenolic acids. The HPLC profiles obtained in the course of this work clearly evidenced a distinct phenolic production between in vivo and in vitro biomass. For example calli and suspended cells produced mainly xanthones while in in vivo plants of both species these compounds were not detected. 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 detected 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. [ABSTRACT FROM AUTHOR]