Fluorometric assay for 1-deoxyfructose
Dills, W.L.; McDonough, G.M.
Analytical Biochemistry 108(2): 325-331
1980
ISSN/ISBN: 0003-2697 PMID: 7457876 Document Number: 159965
A sensitive fluorometric assay was developed for the measurement of 1-deoxyfructose in biological fluids. Samples containing 1-deoxyfructose are incubated with an equal volume of 0.01 M 3,5-diaminobenzoic acid in 5.0 M phosphoric acid in a boiling-water bath for 15 min. The fluorescent product has an emission maximum at 502 nm and an excitation maximum at 396 nm. Fluorescence is proportional to 1-deoxyfructose concentrations over a range from 0.002-1.0 mM. The method can be used to detect as little as 0.03 .mu.mol of 1-deoxyfructose in deproteinized blood and in urine and no interference is observed with glucose, fructose, pyruvate, or ketone bodies. The method will also detect 1-deoxytagastose, 2-deoxyaldohexoses, and -pentoses, 2,5-anhydromannose, and a number of 2-, 3-, 4- and 5-mono- or bis(hydroxymethyl)furans. The fluorescence properties of the products formed from all of the above compounds are similar, suggesting structural similarities of the adducts formed and possible mechanistic similarities of the reactions involved.