Quality-by-Design R&D of a Novel Nanozyme-Based Sensor for Saliva Antioxidant Capacity Evaluation

Oxidative stress is one of the main causes of cell damage, leading to the onset of several diseases, and antioxidants represent a barrier against the production of reactive species. Saliva is receiving increasing interest as a promising biofluid to study the onset of diseases and assess the overall health status of an individual. The antioxidant capacity of saliva can be a useful indicator of the health status of the oral cavity, and it is nowadays evaluated mainly through spectroscopic methods that rely on benchtop machines and liquid reagents. We developed a low-cost screen-printed sensor based on cerium oxide nanoparticles that can be used to assess the antioxidant capacity of biofluids as an alternative to traditional methods. The sensor development process was investigated via a quality-by-design approach to identify the most critical parameters of the process for further optimization. The sensor was tested in the detection of ascorbic acid, which is used as an equivalent in the assessment of overall antioxidant capacity. The LoDs ranged from 0.1147 to 0.3528 mM, while the recoveries varied from 80% to 121.1%, being therefore comparable with those of the golden standard SAT test, whose recovery value was 96.3%. Therefore, the sensor achieved a satisfactory sensitivity and linearity in the range of clinical interest for saliva and was validated against the state-of-the-art equipment for antioxidant capacity evaluation.