FAB CIDMS/MS analysis of partially methylated maltotrioses derived from methylated amylose : a study of the substituent distribution

Abstract

Amylose was methylated with CH3I in alkaline aqueous suspension, yielding methylated amylose (MeAl) with a degree of substitution of 1.44 (s<0.01). Determination of the monomer composition showed that HO-6 and HO-2 were highly substituted in contrast to HO-3 (7:2:5.5, HO-2:HO-3:HO-6). By using partial acid hydrolysis, oligomers were prepared that varied both in degree of polymerisation and in methyl-content. Studies on the distribution of substituents in trimers showed large deviations from random distributions. By using CID tandem mass spectrometry, the substituent distribution in these trimers was determined in more detail. Various sets of trimers with equal amounts of methyl-groups but differing in substituted positions were quantified. From the monomer composition of MeAl, the probability of each trimer was calculated and compared to the outcome of the measured distributions. It was concluded that trimers with terminal tri- or non-substituted glucose monomers at the non-reducing end were formed preferentially during partial hydrolysis and that partial hydrolysis of MeAl yielded oligomers in a non-random way. This is the first study that describes the partial hydrolysis of MeAl in such detail.