Analytical methodology for multidimensional size/branch-length distributions for branched glucose polymers using off-line 2-dimensional size-exclusion chromatography and enzymatic treatment

Vilaplana, Francisco and Gilbert, Robert G. (2011) Analytical methodology for multidimensional size/branch-length distributions for branched glucose polymers using off-line 2-dimensional size-exclusion chromatography and enzymatic treatment. Journal of Chromatography A, 1218 28: 4434-4444. doi:10.1016/j.chroma.2011.05.027


Author Vilaplana, Francisco
Gilbert, Robert G.
Title Analytical methodology for multidimensional size/branch-length distributions for branched glucose polymers using off-line 2-dimensional size-exclusion chromatography and enzymatic treatment
Journal name Journal of Chromatography A   Check publisher's open access policy
ISSN 0021-9673
1873-3778
Publication date 2011-07
Sub-type Article (original research)
DOI 10.1016/j.chroma.2011.05.027
Volume 1218
Issue 28
Start page 4434
End page 4444
Total pages 11
Place of publication Amsterdam, Netherlands
Publisher Elsevier
Collection year 2012
Language eng
Abstract Instrumental and procedural optimizations are developed for a new method to obtain 2-dimensional distributions for branched homopolymers based on size and branching. The method uses 2-dimensional off-line size-exclusion chromatography (SEC) and chemical debranching, in this case using debranching enzymes on branched glucose polymers. This treatment, first presented for the 2D weight and number distributions of starch [F. Vilaplana, R.G. Gilbert, Macromolecules 43 (2010) 7321] is applied here to give 2D weight distributions as functions of size and branch length for glycogen, amylose, and native starch containing both amylopectin and amylose. Completely dissolved polysaccharides are first fractionated by size (hydrodynamic volume) using preparative SEC; the collected fractions are then debranched by addition of isoamylase, and the size distributions of the corresponding branches are analyzed for each fraction using analytical SEC with differential refractive index and with multiple-angle laser light scattering detection. Operational parameters have been optimized to provide sufficient separation resolution for each dimension (size of the whole branched macromolecule and size of the resulting branches after debranching) and to minimize degradation. These 2-dimensional distributions bring out new features in the structure of these branched polysaccharides, and offer a useful tool to elucidate relations between biosynthesis, structure, and properties.
Keyword Size-exclusion chromatography (SEC)
Branched polysaccharides
Size distributions
Branching structure
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2012 Collection
Centre for Nutrition and Food Sciences Publications
 
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