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Molecular Characterization of Barley β-Glucans by Size-Exclusion Chromatography with Multiple-Angle Laser Light Scattering and Other Detectors
Authors:B E Knuckles  W H Yokoyama  M M Chiu
Abstract:Molecular characteristics were determined for mixed-linkage (1→3) (1→4)-β-d -glucans (β-glucans) extracted from Azhul, Crystal, Waxbar, and Prowashonupana barleys. β-Glucans in extracts (with or without α-amylase, protease, hemicellulase, or xylanase treatment) were separated from other components by high-performance size-exclusion chromatography and detected with multiple-angle laser light scattering, refractive index, and fluorometry following postrefractive index treatment with Calcofluor. Pretreatment of barley with 70% ethanol (80°C, 4 hr) reduced β-glucanase activity by ~20%. Hot-alcohol treatment also reduced β-glucan extraction at 23 and 65°C by 42 and 14%, respectively. Molecular weights of β-glucans in the first water extract were generally higher than in succeeding water and alkali extracts. Weight average molecular weights ranged from 0.44 × 106 to 2.34 × 106 g/mol after α-amylase treatment to remove interfering starch. Interference due to pentosans was not demonstrated using enzyme treatments.
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