Ultrasensitive optical detection of hydrogen peroxide by triggered activation of horseradish peroxidase

Hydrogen peroxide is a very reactive byproduct of many metabolic pathways. We describe an ultra-sensitive colorimetric method to detect hydrogen peroxide based on the reconstitution of apo-horseradish peroxidase with the hemin derivative, hemin di(N,N'-acetyl-hydrazide). Oxidation of the latter by hydrogen peroxide yields hemin, which is able to reconstitute apo-horseradish peroxidase, forming an active peroxidase. We have also applied this analyte-triggered reconstitution of horseradish peroxidase to detect the activity of the enzyme glucose oxidase.