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Several methods were used for the characterization of starch granules isolated from barley kernels. A procedure based on a combination of alkaline digestion, toluene treatment and filtration over sieves with pore diameters of 70 and 40 μm was used for isolation and purification of starch granules from kernels. The released starch granules were characterized by various methods: scanning electron microscopy (SEM), image analysis of optical microscopy data (IAOM), low angle laser light scattering (LALLS), and gravitational field‐flow fractionation (GFFF). All methods showed the bimodal size distribution of the isolated starch granules, however, they differed in the ratio of large and small starch granules. LALLS and GFFF were also used for determination of the ratio of large and small starch granules (ratio A/B) isolated from two malting barley cultivars Kompakt and Akcent. Both techniques determined the higher ratio A/B for the cultivar Akcent. SEM was also used to examine the extent of digestion. The micrographs indicate that a significant proportion mainly of small granules are still embedded into residues of endosperm and a more extensive digestion must be performed to release all starch granules from barley kernels.

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A newly developed method GFFF (Gravitational Field-Flow Fractionation) and the well known method LALLS (Low Angle Laser Light Scattering) were used to assess starch granule size distribution of ten varieties of spring barley. As a distribution criterion, the ratio of starch granule content larger than 8 mm (type A) and smaller than 8 mm (type B) was chosen. Both methods divided the observed set in a similar way. Varieties Akcent, Forum and Atribut formed a variety set with the highest ratio of large and small starch granules. Varieties Scarlet and Kompakt had intermediate ratios. The remaining five varieties Amulet, Novum, Olbram, Tolar and Krona had the lowest ratios of large and small starch granules. Statistical analysis showed that there was a highly significant correlation between the GFFF and LALLS methods.