Modification of Standard Enzymatic Protocol to a Cost-Efficient Format for Mixed-Linkage (1→3,1→4)-β-d-Glucan Measurement

The current enzymatic assay approach (AACC International Approved Method 32-23) for the measurement of mixed-linkage beta-glucan in small grains wits modified to a cost-efficient and high-throughput format without compromising the accuracy of the results. Ten barley (Hordeum vulgare L.) genotypes used in the study represented a wide range of beta-glucan content levels. A reduced reaction Volume is used in the new protocol to adapt to a 96-well plate format. The Volume of key components lichenase and beta-glucosidase were reduced to 25% of the volume required in the original protocol and the cost per sample was reduced to 22% of that in the original protocol. Labor cost was also decreased to 25% of the original protocol as a result of format changes. The accuracy of the measurement front the modified protocol was comparable to the current standard enzymatic procedure. P-Glucan measurement accuracy of the modified and original protocols were also compared using 21 oat (Avena sativa L.) samples. The results indicated that the new protocol consistently produced accurate measurements in both barley and oat.