Non-enzymatic colorimetric detection of hydrogen peroxide using a μPAD coupled with a machine learning-based...

In the present study, iodide-mediated 3,3',5,5'-tetramethylbenzidine (TMB)-H₂O₂ reaction system was applied to a microfluidic paper-based analytical device (μPAD) for non-enzymatic colorimetric determination of H₂O₂. The proposed system is portable and incorporates a μPAD with a machine learning-based smartphone app. A smartphone app called "Hi-perox Sens" capable of image capture, cropping and processing was developed to make the system simple and user-friendly. Briefly, circular μPADs were designed and tested with varying concentrations of H₂O₂. Following the color change, the images of the μPADs were taken with four different smartphones under seven different illumination conditions. In order to make the system more robust and adaptive against illumination variation and camera optics, the images were first processed for feature extraction and then used to train machine learning classifiers. According to the results, TMB + KI showed the highest classification accuracy (97.8%) with inter-phone repeatability at t = 30 s under versatile illumination and maintained its accuracy for 10 minutes. In addition, the performance of the system was also comparable to two different commercially available H₂O₂ kits in real samples.