2.6. Identification and qualification of anthocyanins
2.6.1. Preparation of sample extract
The anthocyanins were extracted according to the method reported
by Sarkis, Jaeschke, Tessaro, and Marczak (2013) with some modification.
A 0.5 g was added with 5.0 ml of acidified methanol (methanol:
HCL, 100:1 (v/v)), and then mixed with a vortex mixer for 1 min and
stored overnight at 4 °C and then the supernatant (subsample) was
filtered through a 0.45 μm nylon syringe filter (Whatman, USA) and
injected into the HPLC system for anthocyanin analysis.