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1860849 
Journal Article 
TRITIUM LABELING OF AMINO-SUGARS AT C-2 BY ALKALINE EPIMERIZATION IN TRITIATED-WATER 
Roden, L; Jin, J; Yu, H; Campbell, P 
1995 
Yes 
Glycobiology
ISSN: 0959-6658
EISSN: 1460-2423 
5 
2 
167-173 
English 
7780191 
WOS:A1995QQ10000007 
N-Acetyl-D-[2-3H]glucosamine was synthesized from N-acetyl-D-mannosamine by alkaline 2-epimerization in pyridine containing 3H2O and nickelous acetate. The reaction involves reversible formation of an enol intermediate and therefore also resulted in incorporation of tritium into N-acetylmannosamine. After completed reaction, the two N-acetylhexosamines were separated from other radioactive products and Morgan-Elson chromogens by chromatography on a column of Sephadex G-10, which was eluted with 10% ethanol, and were then separated from each other by chromatography on Sephadex G-15 in 0.27 M sodium borate (pH 7.8). The location of the incorporated tritium was established by treatment of the N-acetylhexosamines with borate under the conditions of the Morgan-Elson reaction, which converts the sugars to Kuhn's chromogen I with concomitant loss of the C-2 hydrogen. As expected, this treatment resulted in the formation of 3H2O, indicating that the tritium was located at C-2. [2-3H]Glucosamine was prepared by acid hydrolysis of the labelled N-acetylglucosamine and was converted to [2-3H]glucosamine 6-phosphate by incubation with hexokinase and ATP. The sugar phosphate was used as a substrate for glucosamine 6-phosphate deaminase (isomerase, EC 5.3.1.10) in a simple 3H2O release assay. 
N-ACETYL-[2-H-3]GLUCOSAMINE; N-ACETYL-[2-H-3]MANNOSIAMINE; [2-H-3]GLUCOSAMINE; GLUCOSAMINE 6-PHOSPHATE DEAMINASE; [2-H-3]MANNOSAMINE