Apigenin is a phytochemical which has been associated with numerous beneficial health effects. However, due to its low concentration and poor bioavailability, isolation from natural sources is very difficult. Subcritical water (SCW) is an excellent solvent for the extraction of moderately polar and non-polar flavonoids. Carefully balanced moderate hydrolytical potential of SCW was used in this work to liberate apigenin from its bound forms. In this research simultaneous extraction and hydrolysis of apigenin bound forms from chamomile have been performed. Optimal operation parameters for maximum yields of apigenin have been investigated and defined (solvent-to-sample ratio 1:30; agitation rate 3 Hz; pressure 45 bar; temperature 115 degrees C; time 30 min). Apigenin was further isolated in its pure form from water extracts obtained under optimal conditions using column chromatography. The purity of isolated compound was confirmed by spectroscopic analysis. Biological activity of extracts obtained under optimal conditions were determined as well. In order to identify polyphenolic compounds UHPLC-HESI-MS/MS analysis was performed. (C) 2016 Elsevier B.V. All rights reserved.