A gravimetric method for the quantitative determination of soluble β-glucan content in oat grain

Abstract

Background. Oat (Avena sativa L.) is one of the most important sources of protein, oil, starch and dietary fibers, in particular β-glucans. Dietary fiber serves as a source of nutrition for the intestinal microflora and significantly affects not only its composition, but also the normal functioning of the intestine as a whole. In connection with the increased interest in β-glucans as a food component and dietary supplement, there is a need in a convenient and inexpensive method for determination of β-glucans content in grain. The article provides an overview of the existing methods for the isolation and determination of soluble β-glucans in cereals: enzymatic, alkaline, alkaline-enzymatic, colorimetric; their advantages and disadvantages are noted. The main disadvantages of some methods are the complexity and duration of execution, significant cost of the required reagents, and the inability to determine the exact content of β-glucans due to their insufficient purification from various impurities.Results. This study used the example of naked and covered oat cultivars to demonstrate applicability of the gravimetric method that we developed by modifying the alkaline method. Whole grain oat flour was pretreated with a 50% ethanol solution to inactivate β-glucanase and remove free sugars, some lipids, proteins, and other substances. β-glucans were released from the aleurone layer and the endosperm with a 5% sodium hydroxide solution, and finally extracted with a 70% ethanol solution. Β-glucans floated to the surface in the form of a bunch of fibers, which was then dried at a temperature about 100-102°C to constant weight and weighed. The content of β-glucans was calculated from the dry weight (%). The isolated β-glucans were checked for the presence of accompanying substances: the content of nitrogenous substances was determined by the Kjeldahl method, and the presence of starch was determined by a qualitative reaction with Lugolʼs reagent.Conclusions. The content of β-glucans in the grain of the studied samples ranged from 3.12±0.18 up to 4.65±0.17% of the dry weight. As a result of the study, the optimal conditions of β-glucans isolation were selected: the extraction mixtures ratios, as well as the modes of centrifugation, sedimentation and drying were established. It has been shown that this method makes it possible to isolate β-glucans with a minimum impurity of nitrogenous substances (0.07-0.12%) and no traces of starch. One of the advantages of the described method is its availability for mass analysis when studying a collection of grain crops.