Thermally Assisted Hydrolysis and Methylation of Purified Tannins from Plants

Abstract

A collection of tannins extracted from various plant species was subjected to thermally assisted hydrolysis and methylation (THM) using tetramethylammonium hydroxide. The products obtained included 1,3,5-trimethoxybenzenes, derived from A rings of condensed tannins (CTs), and 1,2-dimethoxybenzene, 1,2,3-trimethoxybenzene, and derivatives as major products from the B ring. 1,2,4-Tri- and two tetramethoxybenzenes were also detected in most analyses. Correlation analyses revealed that they were derived from the B ring, with 1,2,4-trimethoxybenzene being derived from a procyanidin (PC) B ring and 1,2,3,5-tetramethoxybenzene from a prodelphinidin (PD) ring. Tannins from species that contained both CT and hydrolyzable tannin (HT) produced mainly permethylated gallic acid moieties upon THM, and the products derived from CTs were less abundant. Most likely, the ester-bound HTs are more easily transmethylated than the cleavage and methylation of the C-C and C-O linked CTs. Statistical analyses of the THM products and 13C NMR data of the tannins showed good correlations between the B ring hydroxylation and the di- and trimethoxybenzenes observed. Using the ratio of the methyl esters of 3,4-dimethoxybenzoic acid and 3,4,5-trimethoxybenzoic acid only provided good correlations of the percent PC as well. Furthermore, the 1,3,5-trimethoxybenzenes may serve as good markers of tannins in plant, soil, sediment or other samples as analyzed by THM.