High-performance anion-exchange chromatography with pulsed amperometric detection of linear and branched glucose oligosaccharides
1991
Ammeraal, R.N. | Delgado, G.A. | Tenbarge, F.L. | Friedman, R.B.
Variation of response according to degree of polymerization (d.p.) of linear malto-oligosaccharides was estimated for a high-performance anion-exchange chromatography system with pulsed amperometric detection (h.p.a.e.-p.a.d.). In the range of d.p. 1-20, response per micromol increased as the d.p. increased to about d.p. 14. Beyond this value, response was variable and showed no clear upward trend. Response per microgram decreased with increasing d.p. The effect of alpha-(1 leads to 6) glycosidic linkages on retention times was also investigated with this system using linear pullulan oligosaccharides, as well as enzymatically generated maltosyl or glucosyl branched oligosaccharides from waxy maize starch. Retention times were reduced about 0.5 min for each alpha-(1 leads to 6) glycosidic linkage present. Isomers with single maltosyl branches could not be resolved from those with single glucosyl branches but were separated from alpha-amylase limit dextrins with multiple branching. Identity of the chromatographic peaks was confirmed (i) for the pullulan fragments by comparing to a commercial d.p. 36 standard; (ii) for d.p. 6-14 singly branched maltosyl oligosaccharides by fractionation, pullulanase hydrolysis, and analysis of the resultant products; and (iii) for the alpha-amylase limit dextrins by hydrolysis of a singly branched cyclodextrin, analysis of the products, and comparison to the known starch limit dextrin structures.
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