DETERMINATION OF FUROSINE IN BIOMEDICAL SAMPLES EMPLOYING AN IMPROVED HYDROLYSIS AND HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC TECHNIQUE

Traditionally, the most sensitive and specific determination of non-enzymatic protein glycation has involved an 18-24-h acid hydrolysis in order to generate the compound furosine, which has been detected employing reversed-phase h.p.l.c. In this study, we have reported that significant quantities of furosine can be generated with much shorter hydrolysis times employing a 90-min vapor-phase acid hydrolysis procedure. The furosine generated by vapor-phase hydrolysis is then quantitated by pulsed amperometric detection using anion-exchange high-performance liquid chromatography. Employing this method, we were able to show that furosine generated from acid hydrolysis of purified hepatic membranes in a diabetic and non-diabetic animal model agreed with traditional methods assessing total glycated protein (i.e., boronate affinity methods).