ANALYSES OF ISOMERIC MONO-O-METHYL-D-GLUCOSES, D-GLUCOBIOSES AND D-GLUCOSE MONOPHOSPHATES BY HIGH-PERFORMANCE ANION-EXCHANGE CHROMATOGRAPHY WITH PULSED AMPEROMETRIC DETECTION

To investigate the contribution of each hydroxyl group of D-glucose to retention on pellicular quaternary amine-bonded resins and to the pulsed amperometric detector response, all sets of isomeric mono-O-methyl-D-glucoses and D-glucobioses and three isomeric D-glucose monophosphates were analysed by high-performance anion-exchange chromatography under alkaline conditions with pulsed amperometric detection. The results showed that the reduction of retention time on mono-O-methylation follows the order 2-OH > 3-OH > 6-OH greater-than-or-equal-to 4-OH > 1-OH. Although the elution order of 1-, 3- and 6-phosphates was the same as obtained for the corresponding methyl esters, in the case of glucobioses the elution position was somewhat affected by the configurational contribution and hydrophobic interaction. The pulsed amperometric detector response was little affected by the acidity of each hydroxyl group of D-glucose. The suppression of the pulsed amperometric detector response by substitution of hydroxyl group seemed to be due to the inhibitory effect of the substituent group on the vicinal hydroxyl groups and, as a rule, the degree of suppression caused by 6-O-substitution was the smallest.