Isolation of a hydroxytyrosol-rich extract from olive leaves (Olea Europaea L.) and evaluation of its antioxidant properties and bioactivity

In this research, a phenol extract of high hydroxytyrosol (OLPE) content was obtained from olive leaves ( Olea europaea L.), and subsequently tested under different contexts. The method used to obtain the OLPE basically involved two steps: the use of strongly-acid aqueous steam, generated from 10% HCl (v/v) at 100°C, to directly hydrolyse the native complex phenols from integral olive leaves, and OLPE recovery by liquid–liquid extraction with ethyl acetate. Hydrolysis time was 1 h. Finally, the dried extract was dissolved in distilled water. The OLPE total phenols were determined by Folin–Ciocalteu’s method and by HPLC analysis. Hydroxytyrosol was about 92% of the total phenols present in OLPE, and the yield was about 0.2% on fresh leaves. OLPE showed antioxidant effects on different food lipids and did not inhibit lactic acid bacteria growth; however, it showed cytotoxicity on NIH/3T3 fibroblasts and human umbilical vein endothelial cells at concentrations higher than 0.32 mM (as hydroxytyrosol).