Peroxyl Radical-induced Lipid Peroxidation in Bovine Plasma: Kinetic Parameters for Simultaneous Antioxidant Activity and Capacity Assessments of Foods

Abstract

Conventional chemical antioxidant assays estimate antioxidant capacity but lack physiological relevance and fail to capture the complexity of oxidative processes in biological systems. This study standardised a kinetic bovine plasma lipid oxidation assay (BOPLA) in which AAPH-derived peroxyl radicals induce oxidation of polyunsaturated fatty acids in bovine plasma, providing a biologically relevant model for evaluating food-related antioxidants in a lipoprotein oxidation system. A range of beverages (rosé and white wines, various fruit juices, teas, and coffee) was evaluated using chemical antioxidant assays, nitric oxide inhibition, and egg yolk lipid peroxidation inhibition. AAPH at 3 mM provided an optimal balance between radical flux and assay discrimination, producing stable oxidation kinetics over 120 min while maintaining sufficient dynamic range to distinguish partial and complete antioxidant inhibition. BOPLA parameters, antioxidant capacity, lipid peroxidation inhibition, and oxidation velocity, showed significant correlations with single-electron transfer assays, including CUPRAC, DPPH, Folin–Ciocalteu reducing capacity (FCRC), and iron-reducing capacity (IRAC), supporting the role of polyphenols in suppressing AAPH-driven lipid peroxidation. Ascorbic acid (0.50–250 mg/L) exhibited dose-dependent behaviour with excellent linearity (R2 = 0.9889–0.9997). Pomegranate juice showed the highest antioxidant potential, whereas lime juice concentrate displayed the lowest chemical antioxidant capacity. Importantly, the kinetic framework of BOPLA enabled discrimination of antioxidant behaviour, allowing samples to be classified as fast-acting, sustained, weak, inactive, or pro-oxidant according to their effects on lipid peroxidation kinetics. Overall, these findings highlight BOPLA as a high-throughput kinetic platform for characterising antioxidant activity and capacity in complex food matrices beyond conventional chemical assays.