Quantitation of the glycation intermediate 3-deoxyglucosone by oxidation with rabbit liver oxoaldehyde dehydrogenase to 2-keto-3-deoxygluconic acid followed by high-performance liquid chromatography.
A simple and sensitive method for the detection of 3-deoxyglucosone was developed using oxidation with crude oxoaldehyde dehydrogenase to 2-keto-3-deoxygluconic acid followed by high-performance liquid chromatography (HPLC). Oxoaldehyde dehydrogenase was prepared from rabbit liver and partially characterized. 2-Keto-3-deoxygluconic acid produced from 3-deoxyglucosone by oxoaldehyde dehydrogenase was derivatized with 1,2-diamino-4,5-methylenedioxybenzene, and the fluorescent products were detected and quantitated by HPLC using a solvent containing borate. In the presence of borate, 2-keto-3-deoxygluconic acid was completely separated from N-acetylneuraminic acid. The detection limit of 3-deoxyglucosone was 2.5 pmol/injection (10 microliters) at a signal-to-noise ratio of 3. This method was used to confirm the inhibitory effect of aminoguanidine on glycation.
Fujii
E
E
Department of Internal Medicine, School of Medicine, Kitasato University, Kanagawa, Japan.
Iwase
H
H
Ishii-Karakasa
I
I
Yajima
Y
Y
Hotta
K
K
D000445:Aldehyde Oxidoreductases / Q000737:chemistry*; D000818:Animals; D002417:Cattle; D002645:Chickens; D002851:Chromatography, High Pressure Liquid; D003847:Deoxyglucose / Q000031:analogs & derivatives* / Q000032:analysis; D005942:Gluconates / Q000032:analysis*; D006146:Guanidines / Q000032:analysis; D006863:Hydrogen-Ion Concentration; D008099:Liver / Q000201:enzymology*; D019158:N-Acetylneuraminic Acid; D010084:Oxidation-Reduction; D011817:Rabbits; D051381:Rats; D012794:Sialic Acids / Q000032:analysis; D013050:Spectrometry, Fluorescence; D013552:Swine
D005942:Gluconates; D006146:Guanidines; D012794:Sialic Acids; C002957:2-keto-3-deoxygluconate; D003847:Deoxyglucose; D000445:Aldehyde Oxidoreductases; C021928:2-oxoaldehyde dehydrogenase (NAD+); C016350:3-deoxyglucosone; D019158:N-Acetylneuraminic Acid; C004479:pimagedine