A Rapid and Simple UHPLC- DAD Method for Individual Anthocyanin Analysis: Optimization and Validation for Wild Mediterranean Berries and Fruit Samples

Abstract

Anthocyanins have gained significant attention for their potent colorant capacity and multifaceted biological activities. As a result, the quantification of anthocyanin content in wild berries and the identification of anthocyanin profiles have become increasingly important, as well as the optimization of methods for these determinations. The present study shows a rapid UHPLC-DAD method for identifying and quantifying individual anthocyanins in selected wild berries and fruit samples of Mediterranean origin, namely, Prunus avium, Fragaria vesca, and Vaccinium myrtillus. Moreover, the applicability of the developed method to different food matrices was verified in cultivated berries, specifically, Punica granatum, Fragaria x ananassa, Rubus spp., and Vaccinium corymbosum. The developed method showed great sensitivity (LOD of 23.92 ng/g, LOQ of 79.73 ng/g), precision (RSD below 2.599%), and accuracy (recovery percentages of 97.97 and 98.88%). It successfully quantified individual anthocyanins (delphinidin-3-O-glucoside, cyanidin-3-O-glucoside, cyanidin-3-O-rutinoside, petunidin-3-O-glucoside, pelargodinin-3-O-glucoside, malvidin-3-O-glucoside) across different berries and fruit samples. The developed methodology allows to conduct routine analysis without the need for mass spectrometry. The range of total anthocyanins quantified in wild berries and fruit samples was 0.03 to 7.66 mg/g of powdered extract corresponding to Prunus avium L. and Vaccinium corymbosum L., respectively.