Total polyphenol content and characterisation of the
cocoa extract
Cocoa extract and analytical characterisationwere provided
by Nutrafur S.A (Murcia, Spain). Total polyphenol content
was determined by the FolineCiocalteu colourimetric
method. Briefly, 50 mg of cocoa extract was mixed with
100 mL of water for soluble extracts or 4 mL of
ethanol þ 96 ml of water for insoluble extracts. Then they
were shaken during 15 min and filtered. Absorbance was
measured at 765 nm. The total polyphenol content was
calculated from the calibration curve (1 ppme8 ppm) using
catechin hydrate (SigmaeAldrich C1788) as a standard value
and expressed as milligrams of catechin per 1.4 g.