Separation and analysis of 4'-epimeric UDP-sugars by borate high-performance liquid chromatography.
Separation of UDP-glucose from UDP-galactose, of UDP-N-acetylglucosamine from UDP-N-acetylgalactosamine, of UDP-glucuronate from UDP-galacturonate, or of UDP-glucosamine from UDP-galactosamine was achieved within 10-45 min by isocratic anion-exchange high-performance liquid chromatography (HPLC) using a flow rate of 2 ml/min. The eluants were composed of borate as complex-forming and eluting agent and of glycerol for protection of the alkali-labile silica packing of the column. This borate HPLC was suitable for the analysis of 4'-epimeric UDP-sugars in the range of 2 to 100 nmol. The applicability of this technique was demonstrated by determination of the relative amounts of 4'-epimeric UDP-amino sugars formed in rat liver after administration of D-galactosamine. Since a high salt content of UDP-sugar samples can interfere with borate HPLC, desalting was performed on a 1-ml C18 cartridge using triethylammonium hydrogen carbonate buffer. This procedure enabled the complete separation of various nucleotides from salts within 10 min prior to HPLC.
D000818:Animals; D001888:Boric Acids; D002851:Chromatography, High Pressure Liquid; D005688:Galactosamine / Q000494:pharmacology; D008099:Liver / Q000378:metabolism*; D051381:Rats; D013237:Stereoisomerism; D014529:Uridine / Q000494:pharmacology; D014539:Uridine Diphosphate Sugars / Q000302:isolation & purification*
D001888:Boric Acids; D014539:Uridine Diphosphate Sugars; D005688:Galactosamine; D014529:Uridine