Anthocyanins in the Berries of 123 Interspecific Black Currant Crosses: A Multi-Analytical Approach

High concentrations of anthocyanin pigments and vitamin C are the principal health-promoting constituents in black currant (Ribes spp.) berries. This study aimed to develop a rapid greener protocol—reducing the analytical workload, application of toxic solvents, and processing time—for analysis of anthocyanins in black currants. The influence of several laboratory factors on anthocyanin extraction was assessed (berry size, disintegration method). A rapid and robust RP-HPLC-DAD method was developed and validated for the separation and quantification of anthocyanins across 123 black currant genotypes. The optimized method employing a Kinetex C18 column demonstrated excellent sensitivity and reproducibility, achieving complete separation within 11 min. Extraction using 50% (v/v) ethanol acidified with 0.36 N HCl proved superior to methanol-based solvents. Single extraction produced similar extract concentration to triple extraction; cryogenic grinding enhanced anthocyanin extractability by approximately 25%. Smaller berries within the genotype contained higher anthocyanin levels than larger berries. On average, delphinidin 3-O-glucoside, delphinidin 3-O-rutinoside, cyanidin 3-O-glucoside, and cyanidin 3-O-rutinoside contributed 12.5%, 43.7%, 6.2%, and 35.0% of total anthocyanin content, respectively, and 97.5% of total anthocyanins, ranging from 90.5% to 98.8%. The developed greener chemistry-compliant protocols facilitate rapid, safe, and reliable high-throughput screening of anthocyanin profiles in black currant, useful for breeding programs and analytical workflows.