Kinetic measurement of total amylase and isoamylase activities with a centrifugal analyzer

Abstract

We evaluated a new kinetic assay for α-amylase (Phadebas® IsoAmylase Test), using modified starch as the substrate and a CentrifiChem 400 centrifugal analyzer system. We determined isoamylase activities by using a selective inhibitor. Results were compared with those obtained with the chromogenic Phadebas dyed-starch technique. The method for total amylase appeared to be rapid and precise (CV = 4%) and results correlated well with the chromogenic method. Samples with activities up to 3000 arb. units/L can be analyzed without dilution. Glucose and pyruvate interfere with the assay, but hemoglobin and bilirubin do not. Pancreatic (P) and salivary (S) isoamylase activity can be determined with acceptable precision (CV(p) = 8%, CV(s) = 10%) by an automated routine procedure with commercially available reagents.