Precise Method for the Measurement of Catalase Activity in Honey

An improved method is reported for the determination of catalase activity in honey. We tested different dialysis membranes, dialysis fluid compositions and amounts, dialysis temperatures, sample amounts, and dialysis times. The best results were obtained by dialysis of 7.50 g sample in a cellulose dialysis sack, using two 3 L portions of 0.015M sodium phosphate buffer (pH 7.0) as the dialysis fluid at 4°C for 22 h. As in previous methods, catalase activity was determined on the basis of the rate of disappearance of the substrate, H 2 O 2 , with the H 2 O 2 determined spectrophotometrically at 400 nm in an assay system containing o-dianisidine and peroxidase. Trials indicated that the best solvent for the o-dianisidine was 0.2M sodium phosphate buffer, pH 6.1; the best starting H 2 O 2 concentration was 3mM; the best HCI concentration for stopping the reaction was 6N; and the best sample volume for catalase measurement was 7.0 mL. Precision values (relative standard deviations for analyses of 10 subsamples of each of 3 samples) were high, ranging from 0.48% for samples with high catalase activity to 1.98% for samples with low catalase activity.