Inhibition of α-glucosidase and α-amylase by Spanish extra virgin olive oils: The involvement of bioactive compounds other than oleuropein and hydroxytyrosol

•Determination of antidiabetic and anti-cholinesterase activity of Spanish EVOOs.•Identification and quantification of PhytoPs, in extra virgin olive oil samples.•Determination of flavonoids, carotenoids and fatty acids in EVOO extracts rich in PhytoPs.•First investigation of the effect of the contents of these bioactive compounds on four enzymes. Despite the wide use of extra virgin olive oil (EVOO) to combat several diseases, the antidiabetic and anti-cholinesterase activity of Spanish EVOO have not been assessed. In order to evaluate which compounds are responsible for these activities of five Spanish EVOOs, in addition to flavonoids, we investigated for the first time the effect of the contents of carotenoids, fatty acids (FAs), and phytoprostanes (PhytoPs) on four enzymes: α-glucosidase, α-amylase, acetylcholinesterase, and butyrylcholinesterase. The extracts of these five Spanish EVOOs were found to contain three flavones, three carotenoids, six FAs, and seven classes of PhytoPs. The samples exhibited no in vitro anti-cholinesterase activity but presented strong antidiabetic activity, in the order: ‘Arbequina’≈‘Picual’≈‘Cuquillo’>‘Hojiblanca’>‘Cornicabra’. The samples showed a higher in vitro hypoglycemic effect than individual or mixed standards, possibly due to interaction between multiple identified compounds and/or a very complex multivariate interaction between other factors.