Advancing antioxidant testing: Standardisation of a human plasma oxidation assay and its integration with chemical and cellular protocols

作者：Marcelo Franchin, Bruno Toribio de Lima Xavier, Yuyang Guo, Liping Luo, Kai Wang, Daniel Granato · 发表于：Food Research International · 年份：2025 · DOI：10.1016/j.foodres.2025.116951 · 被引用次数：7 · 研究领域：Free Radicals and Antioxidants、Vitamin C and Antioxidants Research、Antioxidant Activity and Oxidative Stress

Chemical antioxidant methods (CAM) are highly reproducible, but they fail to capture the dynamic nature of biological systems. In contrast, cellular antioxidant assays (CAA) offer a more comprehensive evaluation of antioxidant activity by assessing redox status. However, these assays are time- and resource-intensive. To address these limitations, we standardised and validated a micro-analytical method, the Plasma Oxidation Assay (POA), which utilises human plasma as a probe for Cu 2+ -induced lipoperoxidation to assess the antioxidant activity and capacity of bioactive compounds simultaneously. We analysed various honey samples for their total phenolic content (TPC) and antioxidant capacity using the DPPH free radical scavenging method, ferric-reducing antioxidant power (FRAP), and iron-reducing capacity (IRC). These honey samples were categorised into three groups based on their antioxidant capacity: low, intermediate, and high. These groups were further analysed using the POA, revealing a significant correlation ( r > 0.80, p < 0.05) between CAM and POA, where samples with lower CAM values also exhibited lower antioxidant capacity and activity using the POA parameters. The POA demonstrated limits of detection and quantification at 0.39 and 1.19 mg of ascorbic acid equivalent/L, respectively, with high repeatability (coefficient of variation: 0.58–7.04 %) and accuracy (recovery: 96.4–112 %). Further cellular-based analysis at the cellular level, using mouse-derived hepatocyt...