Isolation of a hydroxytyrosol-rich extract from olive leaves (<Emphasis Type="Italic">Olea Europaea</Emphasis> 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).