Macromolecular Characterization of Three Barley β-Glucan Standards by Size-Exclusion Chromatography Combined With Light Scattering and Viscometry: An Inter-Laboratory Study

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Six (1→3)(1→4)-β-D-glucan standards (A-F) isolated from barley were analyzed by size-exclusion chromatography (SEC) in five different laboratories with varying columns, solvent conditions and detector systems (low- and multi-angle light scattering and viscometry). Static (batch) measurements by capillary viscometry and laser light scattering were included. Fairly consistent results were obtained for the weight average molecular weights (Mw), radii of gyration (RG) and intrinsic viscosities [η], demonstrating that the β-glucans may serve as useful standards or reference materials in the study of cereal β-glucans. Average values for Mw were: A,E: 114,000 (±11%); B,C: 374,000 (±9%), D,F: 228,000 (±13%). Some inconsistencies regarding the polydispersity (Mw/Mn) could be ascribed to the influence of peak broadening in certain column/solvent systems. The study further demonstrated that individual researchers tended to use different processing parameters, especially refractive index increments (dn/dc), due to ambiguities in the literature or to differing experimental values. The need for consistent parameters and processing methods is clearly demonstrated.