Determination of ß-Glucan in Barley and Oats by Streamlined Enzymatic Method: Summary of Collaborative Study
Abstract A collaborative study was conducted involving 8 laboratories (including the authors’ laboratories) to validate the streamlined enzymatic method for determination of ß-D-glucan in barley and oats. In the method, the flour sample is cooked to hydrate and gelatinize ß-glucan, which is subsequently hydrolyzed to soluble fragments with the lichenase enzyme. After volume and pH adjustments and filtration, the solution is treated with ß-glucosidase, which hydrolyzes ß-gluco-oligosaccharides to D-glucose. D-Glucose is measured with glucose oxidase-peroxidase reagent. Other portions of lichenase hydrolysate are treated directly with glucose oxidase-peroxidase reagent to measure free glucose in test sample. If levels of free glucose are high, the sample is extracted first with 80% ethanol. For all samples analyzed, the repeatability relative standard deviation (RSDr) values ranged from 3.1 to 12.3% and the reproducibility relative standard deviation (RSDR) values ranged from 6.6 to 12.3%. The streamlined enzymatic method for determination of ß-D-glucan in barley and oats has been adopted first action by AOAC INTERNATIONAL.