Isolation and identification of the rearrangement products of diflunisal 1-O-acyl glucuronide.

A preparative reversed-phase high-performance liquid chromatographic method is described for the simultaneous separation of eight different isomers formed from the 1-O-acyl glucuronide of diflunisal. All isomers were formed when the acyl glucuronide was incubated under mildly alkaline conditions in aqueous solution. Various forms of two-dimensional NMR studies were performed in order to identify each isomer. Seven of the isomers were identified as alpha- and beta-forms of esters in which diflunisal forms an ester with one of the four alcohol groups in the glucupyranuronic acid. One isomer was identified as the ether glucuronide of diflunisal. To establish the exact chemical shift of the different protons, simulation of the one-dimensional NMR spectra and iterative analyses were performed.