Automated analysis of 2,3-diamino-2,3-dideoxy-D-glucuronic acid by cation exchange chromatography with fluorometric postcolumn derivatization.
2,3-Diamino-2,3-dideoxy-D-glucuronic acid (diaminoglucuronic acid) occurs as its di-N-acetyl derivative as a unique constituent of some bacterial cell walls. A sensitive chromatographic method for its determination is described. Diaminoglucuronic acid was well separated from glucosamine and galactosamine in about 80 min on a Dionex DC-6A cation exchange column (0.9 x 18 cm, 50 degrees C) with a sodium citrate buffer (pH 5.28) containing boric acid (0.2 M). The amino sugars in the eluate were monitored fluorometrically by postcolumn derivatization with orthophthalaldehyde detection reagent. This method allowed the automated determination of 50-100 pmol of glucosamine, galactosamine, and diaminoglucuronic acid and was applied successfully to the analysis of diaminoglucuronic acid in Propionibacterium acnes cells.
Verostek
M F
MF
Department of Biochemistry, Albany Medical College, New York 12208.
Bartholomew
L E
LE
Weber
P
P
D000596:Amino Acids / Q000032:analysis; D001322:Autoanalysis; D002852:Chromatography, Ion Exchange / Q000379:methods; D005965:Glucuronates / Q000032:analysis*; D006595:Hexosamines / Q000032:analysis; D007202:Indicators and Reagents; D013050:Spectrometry, Fluorescence / Q000379:methods
D000596:Amino Acids; D005965:Glucuronates; D006595:Hexosamines; D007202:Indicators and Reagents; C035700:2,3-diamino-2,3-dideoxyglucuronic acid