Continuous-flow determination of an alpha-amylase inactivator in fermentation samples using a ferricyanide reagent.
A continuous-flow system has been developed in which pancreatic alpha-amylase is incubated with soluble starch at 37 degrees C. Reducing sugars being delivered at the "steady-state" hydrolysis of starch are dialyzed into a solution of alkaline ferricyanide. Ferricyanide is then reduced at 95 degrees C. The decrease in the absorbance of ferricyanide solution is recorded as "enzyme baseline." When samples containing alpha-amylase inactivator are introduced into the system, hydrolysis of starch is reduced according to the concentration of enzyme inactivator. The amount of inactivator in the samples is deduced from a standard curve of six standard concentrations using the Technicon Autoanalyzer II system manager in connection with a calculator for automated interpolation. This method allows to determine 40 samples/h with high sensitivity and precision (mean = 1.53 mg/liter; C.V. = 1.4%).
D000818:Animals; D001322:Autoanalysis / Q000295:instrumentation; D005285:Fermentation; D005292:Ferricyanides; D007202:Indicators and Reagents; D010179:Pancreas / Q000201:enzymology; D013552:Swine; D000516:alpha-Amylases / Q000032:analysis / Q000037:antagonists & inhibitors*
D005292:Ferricyanides; D007202:Indicators and Reagents; C007931:hexacyanoferrate III; D000516:alpha-Amylases