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

¹USDA-ARS, Small Grains and Potato Germplasm Research Unit, 1691 South 2700 West, Aberdeen, ID 83210. Names are necessary to report factually on available data; however, the USDA neither guarantees nor warrants the standard of the product, and the use of the name by the USDA implies no approval of the product to the exclusion of others that may also be suitable. ²Corresponding author. Phone: 208-397-4162. Fax: 208-397-4165. E-mail address: gongshe.hu@ars.usda.gov

The current enzymatic assay approach (AACC International Approved Method 32-23) for the measurement of mixed-linkage β-glucan in small grains was 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 β-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 β-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 from the modified protocol was comparable to the current standard enzymatic procedure. β-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.