Extraction and determination of biologically active components of St. John’s wort and its pharmaceutical preparations

Abstract

A procedure is developed for the determination of biologically active substances (BAS) of common St. John’s wort (Hypericum perforatum L.) by HPLC using two columns, Luna C18, 100 Å (for the determination of phenolcarboxylic acids and flavonoids), and Onyx Monolithic C18 (for the determination of phloroglucinols and naphthodianthrones), in the gradient elution mode with diode array detection. The detection limits for analytes are 0.05–0.20 μg/mL. To optimize the conditions, we studied the extraction of biologically active substances from St. John’s wort by a water–ethanol solution at different temperatures and pressures and under the effect of microwave radiation and ultrasound. The maximum amounts of substances were extracted in a dynamic mode under heat and pressure. The procedure was applied to the St. John’s wort samples of different brands and some pharmaceutical products based on it. The components of extracts were identified by retention times, UV spectra, and mass spectra. It was shown that the content of biologically active substances in pharmaceutical samples of St. John’s wort depends on the herb habitat. It was shown that hyperforin decomposed in pharmaceutical formulations based on St. John’s wort during storage.