Determination of Glucose Oxidase Activity by Tetramethylbenzidine Chromophotometry Method
In this work,the activity of glucose oxidase was determined by tetramethylbenzidine(TMB)catalyzed fluorescence method.Under the conditions of acetate buffer(pH 4.5)and water bath(37 ℃),glucose under the action of glucose oxidase was quantitatively converted to H2O2,and H2O2 oxidized TMB to produce TMB oxidation products(TMBox)with strong fluorescence signal when Fe2+ions existed.The fluorescence intensity of TMBox was related to the activity of glucose oxidase.The analysis process was divided into three stages including creation of the H2O2 standard curve equation,achievement of enzyme catalyzed reactions and TMB fluorescence reactions,and measurement of the fluorescence intensity of TMBox.The results showed that the detection range of glucose oxidase activity was 3.33×10-5-3.33×10-3 U/mL with detection limit of 1.11×10-6 U/mL.The activity of glucose oxidase in food additives was also determined by tetramethylbenzidine catalytic fluorescence method,and the recovery was between 94.60％and 102.0％.
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