Abstract:
The present study focused on the development of a new purification protocol suitable for betanin and other
major betalains, vulgaxanthin I, indicaxanthin and neobetanin, using flash chromatography which is a conve nient and fast method to isolate unstable materials. Following preliminary tests, a gradient procedure using
0–60% acetonitrile, with 0.1% (v/v) formic acid as mobile phase, was selected for the purification. Different
fractions were collected based on UV detection at 254 and 280 nm and purities were confirmed by reverse-phase
HPLC analysis to be 97%, 95%, 79% and 52% for betanin, indicaxanthin, vulgaxanthin I, and neobetanin,
respectively, with pigment yields ranging from 120 to 487 mg per 100 g of powdered raw material. Comparative
assessment of antioxidant and radial scavenging properties of individual betalains indicated highest potential for
betanin followed by neobetanin, vulgaxanthin I and indicaxanthin.