The linear gradient started with 1% B, keeping isocratic conditions during 5 min, reaching 20% B at 20 min, 40% B at 30 min, 95% B at 35 min and 1% B after 41 min. UV chromatograms were recorded at 520 nm and compared with those obtained with analytical stan-dards. Anthocyanins were quantified by the area of their corre-sponding peaks as Cy3G at 520 nm.