Quantification in patient urine samples of felbamate and three metabolites: Acid carbamate and two mercapturic acids

Purpose: Previously we proposed and provided evidence for the metabolic pat hway of felbamate (FBM), which leads to the reactive metabolite, 3-carbamoy l-2-phenylpropionaldehyde. This aldehyde carbamate was suggested to be the reactive intermediate in the oxidation of 2-phenyl-1,3-propanediol monocarb amate to the major human metabolite 3-carbamoyl-2-phenylpropionic acid. In addition, the aldehyde carbamate was found to undergo spontaneous eliminati on to 2-phenylpropenal, commonly known as atropaldehyde. Moreover, atropald ehyde was proposed to play a role in the development of toxicity during FBM therapy. Evidence for atropaldehyde formation in vivo was reported with th e identification of modified N-acetyl-cysteine conjugates of atropaldehyde in both human and rat urine after FBM administration. Identification of the atropaldehyde-derived mercapturic acids in urine after FBM administration is consistent with the hypothesis that atropaldehyde is formed in vivo and that it reacts with thiol nucleophiles. Based on the hypothesis that the po tential for toxicity will correlate to the amount of atropaldehyde formed, we sought to develop an analytic method that would quantify the amount of r elevant metabolites excreted in patient urine. Methods: We summarize the results of an LC/MS method used to quantify FBM, 3-carbamoyl-2-phenylpropionic acid and two atropaldehyde-derived mercapturi c acids in the patient population. Results: Analysis was performed on 31 patients undergoing FBM therapy. The absolute quantities of FBM and three metabolites were measured. Conclusions: This method demonstrated sufficient precision for the identifi cation of patients exhibiting "abnormal" levels of atropaldehyde conjugates and may hold potential for patient monitoring.