Rapid Screening of Antioxidant Anthocyanins in Autochthonous Nero d’Avola Grape Clones by Pre-column DPPH Reaction Coupled to UHPLC-UV/Vis-IT-TOF: a Strategy to Combine Chemical data and Genetic Diversity

Grape is a rich source of bioactive compounds; among them anthocyanins are associated to many healthy properties, possessing a good antioxidant activity. In this work, we developed a fast and simple screening of antioxidant anthocyanins in six Sicilian Nero d’Avola autochthonous grape clones. The method was based on the pre-column reaction with 2,2′-diphenyl-1-picrylhydrazyl radical followed by the rapid separation by ultra high-pressure liquid chromatography coupled with ultraviolet and tandem mass spectrometry detection. Peak areas of antioxidant anthocyanins significantly reduced or even disappeared. The entire method took only 45 min per sample, showing good retention time and peak area repeatability with maximum CV% values ≤0.86 and 6.84, respectively. Samples rich in delphinidin derivatives showed lowest IC 50 values, since those compounds possess the highest scavenging ability. The developed setup was less complex than online approaches and faster with respect to conventional high-performance liquid chromatography methods, taking advantage of ultra high-performance conditions, coupled for the first time with pre-column 2,2′-diphenyl-1-picrylhydrazyl assay. The proposed strategy is a valid tool for a rapid screening of antioxidant anthocyanins in grape samples, useful to correlate the genetic diversity with the production of secondary metabolites as well to assess their activity in nutraceutical products rich in anthocyanins.