Purification, Fractionation and Identification of Proanthocyanidins from Lotus (Nelumbo nucifera Gaertn.) Seedpods

Abstract

Abstract: Lotus proanthocyanidin extract (LSPE) from lotus seedpods was prepared through extraction with aqueous methanol and purification by macroporous resin adsorption. Average degree of polymerization of LSPE was measured by means of acid degradation. Moreover, it was fractionated using TSK HW-40s resin and analyzed by high performance liquid chromatography (HPLC) with diode array detection and electrospray ionization-mass spectrometry (ESI-MS). The results showed that the extraction yield of proanthocyanidins was 7.65% after 1 h extraction at 60 ℃ with 60% aqueous methanol solution acidified to pH 2.0 at a solid-to-solvent ratio of 1:20 (m/V). AB-8 resin was the most suitable for LSPE purification among three types marcroporous resin investigated. The average degree of polymerization of LSPE was 3.2 in the ethyl acetate phase and 15.4 in the water phase. The ethyl acetate-soluble fraction of LSPE mainly comprised catechin, gallocatechin, and their dimers, trimers and tetramers in small quantities. This fraction was further fractionated by TSK HW-40s resin chromatography into 11 sub-fractions (F1—F11). F1, F2, F3 and F4 might be flavonoids or flavonoid glycosides, F5 was mainly composed of catechin and a small amount of epicatechin, F6 was made up of gallocatechin, catechin, procyanidins dimmer B3 and proanthocyanidins containing gallocatechin units, F7 consisted of catechin, procyanidins dimmers B1, B2 and B3, F8 comprised proanthocyanidins, and F9, F10 and F11 was likely to be polymers with higher degree of polymerization consisting of catechin and gallocatechin.

Key words: lotus, proanthocyanidin, fractionation, identification, electrospray mass spectrometry (ESI-MS)

References

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