Cefalu W T, Bell-Farrow A, Wang Z Q, Ralapati S
Department of Medicine, Bowman Gray School of Medicine, Wake Forest University, Winston-Salem, NC.
Carbohydr Res. 1991 Aug 12;215(1):117-25. doi: 10.1016/0008-6215(91)84012-4.
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).
