Next-generation analytical platforms for antioxidant capacity assessment:  The urge for realistic and physiologically relevant methods

Bioactive compounds, such as carotenoids, alkaloids, and phenolics, are well known because of their alleged  health benefits when consumed regularly in a balanced healthy diet. Some well-documented bioactivities are  antioxidant, antihypertensive, antihyperglycemic, antilipidemic, anti-obesity, anti-inflammatory, and antimicrobial capacities. Trying to associate the chemical composition of distinct sources and their bioactivity using in  vitro methods, several assays have been developed, implemented, and optimised to recapitulate human physiological conditions. However, in most cases, pitfalls are apparent, and no single test tube-based assay can predict in vivo responses. The need for a more physiologically relevant cell-based method to evaluate the antioxidant  capacity of putative antioxidants is apparent. Therefore, in this Review, the current state-of-the-art in food  science and nutrition is aligned with cell biology/bioengineering approaches to propose combining in vitro  digestion and absorption to obtain a bioavailable fraction containing antioxidants. Overall, human plasma, 2-  dimensional human cell lines, such as erythrocytes, lymphocytes, hepatocytes, enterocytes and, ultimately, 3-  dimensional spheroids (organoids) could be used as biologically relevant models to assess the antioxidant activity of compounds, foods, and nutraceuticals. This versatile approach is deemed suitable, accurate, reproducible, and physiologically relevant to evaluate the protective effects of antioxidants against ROS-mediated  oxidation in vitro.