Determination of the contents of A- and B-starches in barley using Low Angle Laser Light Scattering

A method was proposed for the determination of the contents of A- and B-starches in barley and for the use in plant breeding stations and brewing/malting laboratories. A combination was used of classical methods (incl. crushing of barley kernels by a roll crusher, steeping in 0.02M HCl, repeated rubbing and filtering through the sieve of 0.08 mm), and novel pieces of know-how (treatment with β-glucanase and cellulase, alkalisation at pH = 10.0 and centrifugation of crude starch suspension through the layer of CsCl). In this way, barley starch of high purity was obtained at a very low loss, using Low Angle Laser Light Scattering for the determination of the size distribution of starch granules. The boundary of the particle size between the peaks of A-starch and B-starch (7 µm) was evaluated from the distribution curves, while the contents of A- and B-starches were calculated from the cumulative curves. As a whole, the method was internally validated and for its repeatability and uncertainty of measurement the stimates of standard deviation s = 1.56 and confidence interval L_1,2 = x± 1.9% were established, respectively. Taking into account the generally well known difficulties associated with the extraction of substances from biological materials, in this case the separation of starch from barley kernels, we believe that the proposed method will bring satisfactory results in practice.