A widely used spectrophotometric assay to quantify olive oil biophenols according to the health claim (EU Reg. 432/2012)

The purpose of this work was to find a simple, cheap, and suitable method, among the most widely employed, able to guarantee a proper determination and quantification of the phenolic content of extra virgin olive oils (EVOOs), in order to satisfy the requirements of the specific health claim (EU Reg. 432/2012). Total phenolic content by Folin–Ciocalteu (FC) was used and compared versus phenolic profile by HPLC-UV, considering this latter as the most sensitive and specific method for evaluating the phenolic content. Both protocols were performed before and after an acid hydrolysis of the polar phenolic fraction that involves a break of the bound forms of hydroxytyrosol (HTyr) and tyrosol (Tyr), with a simplification of the phenolic profile, and quantification of their total free forms. Results of the phenolic compounds of twelve EVOOs, determined by the different analytical approaches, were statistically compared by means of two-tailed paired t-tests: data obtained by the FC assay (expressed as HTyr) before and/or after acid hydrolysis were statistically comparable with results obtained by acid hydrolysis-HPLC (as sum of HTyr and Tyr).