HPLC analysis of hexosamine phosphates in biological samples.
Galactosamine is quickly metabolized to galactosamine 1-phosphate in rats treated with this compound. An HPLC method to quantify hexosamine phosphates in biological samples is described, modified from the o-phthaldialdehyde amino acid analysis procedure. o-Phthaldialdehyde derivatives of hexosamines and hexosamine-phosphates can be eluted from a reverse-phase column at different retention times, with a total analysis time of 30 min and without overlapping with free amino acids at physiological concentrations. The standard curves are linear between 1 and 40 nmol. This simple method is more selective and sensitive than previous enzymatic analyses of hexosamine phosphorylation.
Ambrosio
S
S
Unitat de Bioquímica, Facultat Odontologia, Universitat de Barcelona, Spain.
De Oliveira
J R
JR
Bartrons
R
R
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005688:Galactosamine / Q000031:analogs & derivatives / Q000032:analysis; D005695:Galactosephosphates / Q000032:analysis; D005944:Glucosamine / Q000031:analogs & derivatives / Q000032:analysis; D005958:Glucosephosphates / Q000032:analysis; D006595:Hexosamines / Q000032:analysis*; D008099:Liver / Q000737:chemistry / Q000166:cytology; D008297:Male; D010710:Phosphates / Q000032:analysis*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013403:Sugar Phosphates / Q000032:analysis*
D005695:Galactosephosphates; D005958:Glucosephosphates; D006595:Hexosamines; D010710:Phosphates; D013403:Sugar Phosphates; C038515:glucosamine 1-phosphate; C079528:galactosamine 1-phosphate; D005688:Galactosamine; D005944:Glucosamine