Direct detection of the internal acyl migration reactions of benzoic acid 1-O-acylglucuronide by C-13-labeling and nuclear magnetic resonance spectroscopy

1-O-Acyl-beta-D-glucopyranuronates can undergo irreversible binding to proteins mainly through internal acyl migration reactions, which may have toxicological significance. A new method based on the C-13-labeling and nuclear magnetic resonance (NMR) spectroscopy has been developed to study the reactivity of the 1-O-acyl-beta-D-glucopyranuronate of benzoic acid. In phosphate buffer (pH 7.4) solution at 37 degrees C, the glucuronide showed apparent first-order degradation kinetics (T-1/2, 125 min), and concurrent and sequential appearance of 2-, 3- and 4-O-acyl isomers as both alpha- and beta-anomers was observed. The isomeric glucuronides were identified by two-dimensional NMR of the reaction mixture. The direct approach using C-13-labeling and NMR could also provide insights into the reactivities of other labile drug acylglucuronides and their isomeric glucuronides. (C) 1997 Elsevier Science Inc.