Qualitative analysis and HPLC isolation and identification of procyanidins from vicia faba

Abstract

The soluble proanthocyanidins of the coloured seed coats of Vicia faba L. were isolated and separated by solvent partition. The chemical characteristics of the proanthocyanidins were elucidated by total oxidation and partial degradation in the presence of phloroglucinol followed by HPLC analysis. The native extract of proanthocyanidins contained (+)-gallocatechin, (-)-epigallocatechin, (+)-catechin and (-)-epicatechin units. Oligomeric procyanidins were purified by chromatography on Sephadex LH-20 and the accessible compounds were isolated by RP-HPLC using a Licrospher Li 100 Column. The structures of the purified oligomeric procyanidins were elucidated using a procedure involving TLC, UV spectroscopy, ESI-MS and HPLC analysis of the products from the phloroglucinol reaction. The major condensed tannins of Vicia faba comprise six compounds identified as two A-type procyanidin dimers, the procyanidin dimers B1, B2 and B3, and a procyanidin trimer.

Flavonoid diversity and biosynthesis in seed of Arabidopsis thaliana

“…Catechin monomer that has a distinct retention time from epicatechin monomer was also not found either in mature seed or during seed development when large amounts of epicatechin were synthesized. These observations are consistent with recent results suggesting that Arabidopsis seed, unlike many other seeds such as grape (Ricardo Da Silva et al 1991), lentil (Duenas et al 2003), Vicia faba (Merghem et al 2004) or soybean (Takahata et al 2001), would exclusively use epicatechin as the building block of PAs (Abrahams et al 2003;Xie et al 2003). The mDP of soluble Ws-2 Arabidopsis seed PAs is calculated to be 5.…”

Characterization of proanthocyanidins in pea (Pisum sativum L.), lentil (Lens culinaris L.), and faba bean (Vicia faba L.) seeds

“…3B; Table 2). Similar PA subunit composition was observed for the cultivar 'Alfred' (Merghem, Jay, Brun, & Voirin, 2004); however, in 'Alfred', catechin (2, 3-transflavan-3-ol) was the most abundant PA extension subunit (Merghem et al, 2004), while in 'Fatima' epigallocatechin (2, 3-cis- Table 1 Reverse-phase HPLC-DAD retention times and characteristic UV/Vis maxima of standards and pea ('CDC Acer'), lentil ('CDC LeMay'), and faba bean ('Fatima') seed PA phloroglucinolysis products. GC-P, gallocatechin-(4α → 2)-phloroglucinol; GC, gallocatechin; EGC-P, epigallocatechin-(4β → 2)-phloroglucinol; EGC, epigallocatechin; C-P, catechin-(4 α → 2)-phloroglucinol; C, catechin; EC-P, epicatechin-(4β → 2)-phloroglucinol; EC, epicatechin.…”

Condensed Tannin Composition Analysis in Persimmon (Diospyros kaki Thunb.) Fruit by Acid Catalysis in the Presence of Excess Phloroglucinol

“…Identification of the CT components in persimmon fruit with phloroglucinol HPLC analysis of five flavan 3-ols and flavan 3-ols gallates phloroglucinol adducts (C-P, EC-P, GC-P, EGC-P, and ECG-P) has been reported in grapevine (Kennedy and Jones, 2001) and pea (Merghem et al, 2004). Based on these reports, we preliminarily analyzed persimmon fruit CTs following acid catalysis in the presence of excess phloroglucinol under the same HPLC conditions but found that some peaks overlapped under these conditions.…”

“…The separated component was collected with TLC, dissolved with 100% MeOH and filtered through a 0.45 μm filter. After concentrating the filtrate under reduced pressure at 35°C, a crude and unidentified component was separated again by TLC developed with toluene : acetone : formic acid (6 : 6 : 1; v/v/v), according to a previous report by Merghem et al (2004). The isolated component was lyophilized and characterized by 1 H-NMR Varian Inova 300 (300 MHz; Varian Inc., Palo Alto, USA), 2D NMR (H-H COSY), and MALDI-TOF MS Bruker Reflux III (Bruker Corporation, Yokohama, Japan) according to the previous report by Zhang and Lin (2008).…”