[Determination of fructose-1,6-diphosphate with aldolase-DNPH by the colorimetric method].
This paper presents a modified method of enzymatic assay for Fructose-1,6-diphosphate(FDP). FDP is split to dihydroxyacetone phosphate (DAP) and glyceraldehyde-3-phosphate (GAP) by the action of aldolase. DAP is hydrolyzed at room temperature to free triose. Under alkaline conditions, the free triose is reacted with 2,4-dinitrophenylhydrazine (DNPH), yielding a 2,4-dinitrophenylhydrazine derivative which dissolve in alkali forming a purple color mixture, with maximum absorption at 540.nm. It is proportional to the contents of FDP. Because the method depends on the colorimetric determination of triose formed from fructose-1,6-diphosphate only by aldolase, glycerophosphate dehydrogenase/triosephosphate isomerase (GDH/TIM) and reduced nicotinamide adenine dinucleotide (NADH) which usually applied in multienzymatic method, are omitted in the modified method. The method is specific, convenient and accuracy for the determination of FDP.
Wang
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Department of Biomedical Engineering, Chongqing University of Medical Sciences, Chongqing 400016.
Xu
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Yu
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Pan
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L
D003124:Colorimetry / Q000379:methods*; D005634:Fructose-Bisphosphate Aldolase / Q000494:pharmacology; D005635:Fructosediphosphates / Q000032:analysis*; D006834:Hydrazines / Q000494:pharmacology
D005635:Fructosediphosphates; D006834:Hydrazines; C446799:dinitrophenylhydrazine; D005634:Fructose-Bisphosphate Aldolase; C029063:fructose-1,6-diphosphate