Abstract
Four anthocyanin pigments in the seed coat of Phaseolus vulgaris, cultiva. Kurodanekinugasa were found by the paper chromatographic test. For the isolation and the identification of these anthocyanin pigments, the seed coat was treated as follows.
Seed coat was macerated with 0.5% methanolic hydrochloric acid and the extracts were concentrated to dryness in vacuo. The water soluble fraction of the extracts was applied to Amberlite CG-50 column. The pigments were eluted from the resins with 0.5% methanolic hydrochloric acid. After concentration of the eluates, four pigments were separated from each other by cellulose powder column chromatography. These pigments were purified by mass paper chromatography and each pigment (A, B, C and D) was crystallized from 1% methanolic hydrochloric acid.
Methoxyl group was found in the three pigments (B, C and D) by Micro Zeisel method. The aglycone and the sugar obtained by acid hydrolysis of each anthocyanin were identified by paper chromatography with authentic specimens. The position and number of sugar residues in the anthocyanins were identified by the partial acid hydrolysis method. And the visible absorption spectra of the anthocyanins were measured.
By these analytical experiments, four anthocyanins were identified as delphinidin-3-monoglucoside, petunidin-3-monoglucoside, malvidin-3-monoglucoside and malvidin-3, 5-diglucoside.
