2.6. 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 μL was mixed with 1700 μL of distilled water
and 200 μL 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. This reading corresponds to the total anthocyanin content
after considering the relevant dilution. Calculations of total anthocyanins
were based on malvidin-3-glucosid (molar absorptivity 28,000).