Characterization of Anthocyanins from Ajuga pyramidalis Metallica Crispa Cell Cultures

The anthocyanins from Ajuga pyramidalis Metallica Crispa cell cultures were characterized by a combination of HPLC, chemical hydrolysis, MS, and 1H NMR. Chromatographic analysis of the in vivo and in vitro samples showed the same suite of anthocyanins and relative composition. Chemical hydrolysis and mass spectral analysis of the major pigment from suspension cultures showed the presence of cyanidin, three glucoses, two ferulic acids, and malonic acid. The structure of the major pigment in the suspension cultures was determined to be 3-O-(6-O(E)-ferulyl)-2-O-[(6-O(E)-ferulyl)-β-d-glucopyranosyl-β-d-glucopyranosyl]-5-O-(6-O-malonyl)-β-d-glucopyranosylcyanidin. The anthocyanins in the in vitro extract were more stable toward light compared to those in the in vivo extract. The initial degradation of the major pigment both from in vitro and in vivo sources was mainly due to demalonylation. Keywords: Ajuga; anthocyanins; cell culture; stability-anthocyanins