Optimization of HPLC method for the isolation of Hypericum perforatum L. methanol extract

Stamenković, J., Radojković, I., Đorđević, A., Jovanović, O., Petrović, G., Stojanović, G.: Optimization of HPLC method for the isolation of Hypericum perforatum L. methanol extract. Biologica Nyssana, 4 (12), December 2013: 81-85. St. John's Wort (Hypericum perforatum L.) is one of the most studied plant species in the family Hypericaceae. The aim of this study was the identification of the constituents of methanol extract of H. perforatum and optimization of conditions for their isolation. The main components of the methanol extract were isolated on preparative ZORBAX Eclipse XDB C18 column with solvent system consisting of methanol and 1x10 -2 M ammonium acetate in water. Constituents of the extract were identified by comparing their retention times with the retention times of the standards, with the literature data and the UV spectra. By varying the conditions of chromatography, the optimal conditions for isolation of the methanol extract constituents were determined: mobile phase consisting of methanol and 1x10 -2 M ammonium acetate in water in ratio 1 : 1, sample concentration 100 mg/mL, sample volume 30 μL, flow 2 mL/min. Under these conditions 7 components of the methanol extract were isolated.