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). © 1991.
Cefalu, W. T., Bell-Farrow, A., Wang, Z. Q., & Ralapati, S. (1991). Determination of furosine in biomedical samples employing an improved hydrolysis and high-performance liquid chromatographic technique. Carbohydrate Research, 215(1), 117–125. https://doi.org/10.1016/0008-6215(91)84012-4