Characterization of proanthocyanidin in hot water extract isolated from Pinus radiata bark

Proanthocyanidins (PAs) in the hot water extract prepared from Pinus radiata bark were isolated by a Sephadex LH-20 chromatography. The PAs were characterized using UV/Visible, FT-IR and C-13 nuclear magnetic resonance (NMR) spectroscopic techniques, matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry (MS), and gel permeation chromatography (GPC) analysis. Bate-Smith acidic alcoholysis and FT-IR spectrum of the PAs indicated the predominant presence of the procyanidin (PC) structure. The C-13 NMR spectrum revealed that the PAs comprise PC (94%) together with a small amount of prodelphinidin (PD, 6%), and also that they contain the higher trans configuration rather than the cis configuration of C2-C3. The MALDI-TOF MS results obviously proved the presence of PD units, but the maximum degree of polymerization (DP) was limited to octamer. The average molar mass (Mw, Mn), polydispersity (Mw/Mn), and DP of the pine bark PAs were 3,800 (Mw), 1,200 (Mn), 3.2, and 13, respectively.