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).
Cefalu
W T
WT
Department of Medicine, Bowman Gray School of Medicine, Wake Forest University, Winston-Salem, NC.
Bell-Farrow
A
A
Wang
Z Q
ZQ
Ralapati
S
S
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003921:Diabetes Mellitus, Experimental / Q000378:metabolism; D006031:Glycosylation; D006868:Hydrolysis; D008099:Liver / Q000737:chemistry; D008239:Lysine / Q000031:analogs & derivatives* / Q000032:analysis; D008562:Membrane Glycoproteins / Q000032:analysis; D051381:Rats; D011919:Rats, Inbred Strains; D013997:Time Factors
D008562:Membrane Glycoproteins; C018948:furosine; D008239:Lysine