A Smartphone Colorimetric Development with TMB/H2O2/HRP Reaction System for Hydrogen Peroxide Detection and its Applications

The residue and overproduction of hydrogen peroxide (H2O2) can harm cellular dysfunction under oxidative stress conditions. The development of a smartphone colorimetric biosensor system and the TMB/H2O2/HRP system aims to quantify H2O2. The TMB/H2O2/HRP system revealed remarkable activity in catalyzing the reaction of the chromogenic substrate 3,3′,5,5-tetramethylbenzidine (TMB) and natural horseradish peroxidase (HRP) with H2O2, producing an oxidized product with an absorption at 450 nm. Under the optimized conditions, an excellent linear correlation between different absorbance values and H2O2 concentration in the range of 50–800 μM was obtained by UV-Vis spectrophotometer (R2 = 0.9964) and a smartphone device (R2 = 0.9959). For quantitative detection, a smartphone colorimetric reading device was used to record the RGB value via the Android application. A prospective portable platform for H2O2 measurement via a smartphone application was successfully used to determine H2O2 levels in beverages with recoveries ranging from 99.66 to 134.67%.