Method for determination of hydrogen peroxide, with its application illustrated by glucose assay

We describe a simple colorimetric method for determining micromolar quantities of hydrogen peroxide, based on the oxidation of iodide in the presence of ammonium molybdate and photometry of the resulting blue starch-iodine complex. Color development is linearly dependent on analyte concentration, but only slightly time dependent, and the color of the complex formed is stable for several hours. In the range of wavelengths that may be used (570 to 630 nm), lack of interference from other biological compounds makes this method seem suitable for routine analyses. As one illustrative application of the method we quantitated glucose by measuring hydrogen peroxide produced from it by glucose oxidase catalysis. This method of quantitating glucose is more than five times as sensitive as the commonly used dianisidine method. With the appropriate hydrogen peroxide-producing oxidases, this method may be used to directly measure amino acids, purines, uric acid, xanthine, and hypoxanthine.