Fast, Multiple-Use Optical Biosensor for Point-of-Care Glucose Detection with Mobile Devices Based on Bienzyme Cascade Supported on Polyamide 6 Microparticles

Non-invasive glucose determination provides major advantages in health monitoring and protection. It enables widespread point-of-care testing, which is affordable, sensitive, specific, rapid and equipment-free. This work reports on the development and analytical performance of a colorimetric biosensor in detecting glucose in human urine. Highly porous polyamide microparticles were synthesized as the support for the glucose oxidase (GOx) and horseradish peroxidase (HRP) dyad, which was immobilized randomly or consecutively—first HRP and then GOx. The latter system was superior, as GH@PA-C showed much higher activity than the random system, and it was used to prepare the biosensor, along with the 3,3′,5,5′-tetramethylbenzidine chromogen. When in contact with urine, the biosensor displayed a strict linear correlation between the color difference and the glucose concentration in urine in the range of 0.01–3.0 mM, as established by the CIELab image processing algorithm and UV-VIS measurements. The biosensor acted in 20 s and had a detection limit of 30.7 µM in urine, high operational activity at pH = 4–8 and unchanged detection performance after 30 days of storage. Its unique feature is the possibility of multiple consecutive uses without the serious deterioration of the recovery and dispersion values. These characteristics can open the way for new routines in non-invasive personal diabetes detection.