Hydrogen peroxide can react with ammonium molybdate to form a yellow complex which has a maximum absorption peak at 405nm. H2O2 content can be calculated by measuring the absorbance value at 405 nm.

1.    Since hydrogen peroxide is unstable, it is necessary to calibrate the actual concentration of hydrogen peroxide before use, dilute reagent 3 (about 1 mol/L) for 100 times, so that the concentration of hydrogen peroxide is about 10 mmol/L, then measure the OD value(A240) at 240 nm.

The actual concentration of Reagent 3 (mmol/L) = 22.94 × A240 × 100 ÷ 1000.

Calculate the actual concentration of the hydrogen peroxide standard provided by this kit and then dilute the hydrogen peroxide standard to 60 mmol/L based on the actual measured concentration.

2.    If the concentration of H2O2 in the sample is too high, please dilute the samples appropriately. If the concentration is too low, the sampling volume of the sample should be increased, and the sampling volume of standard and double distilled water should be increased equally at the same time.

1.   Add 1 mL of reagent 1 to 5 mL EP tube and preheat at 37℃ for 10 min.

2.   Blank tube: add 0.1 mL of double distilled water to 5 mL EP tube.

Standard tube: add 0.1 mL of 60 mmol/L H2O2 standard solution to 5 mL EP tube.

Sample tube: add 0.1 mL of sample to 5 mL EP tube.

3.   Add 1 mL of reagent 2 and mix fully with vortex mixer.

4.   Set spectrophotometer to zero with double-distilled water, then measure the OD values of each tube at 405 nm with with 1 cm optical path quartz cuvette.