Technical note: a simple, rapid assay for alpha-amylase in bovine pancreatic juice

A procedure is described for the determination of alpha-amylase activity in bovine pancreatic juice. Bovine pancreatic juice was incubated with potato amylopectin for 6 min, and then alkaline potassium ferricyanide was added to stop the enzymatic reaction. Tubes were placed in a boiling water bath for 10 min and then cooled, and 40 mL of distilled water was added. Optical density was read at 420 nm, and the amount of end product (reducing sugars) was determined by comparison with maltose standards. Ten dilutions of bovine pancreatic secretion were assayed to determine the intraassay precision of this method, resulting in a CV of 1.05%. A pooled sample was assayed five times over five separate days to determine the interassay precision, resulting in a CV of 7.3%. This alpha-amylase assay provides a repeatable, rapid, and economical determination of alpha-amylase activity from bovine pancreatic juice.