2.4.4. Total anthocyanins
Total anthocyanins were determined using a modified method of Mazza, Fukumoto, Delaquis, Girard, and Ewert (1999). A 10-fold diluted sample of 100 lL was mixed with 1700 lL of distilled water and 200 lL of 5% (v/v) HCl. The sample was hold at room temperature for 20 min before measuring the absorbance at 520 nm in a 10 mm cuvette. Calculations of total anthocyanins were based on cyanidin-3-glucoside (molar absorptivity 25,740). All spectrophotometric analyses were performed using a UV–visible spectrophotometer Lambda 20 (Perkin–Elmer, Überlingen,Germany).