Lipid oxidation is the main hurdle for omega-3 fatty acid enrichment in food and beverages. Fat oxidation reduces the quality and safety of supplemented products. A tuna oil-in-water emulsion (20%v/v) was exposed to iron-induced oxidation. Emulsions with changing emulsifiers and buffers were analyzed under different storage conditions (argon purging, pH variation) using Conjugated Dienes and Thiobarbituric acid reactive substances assays. The results showed that free iron ions cannot interact with oxygen. However, buffers (Citrate and phosphate) chelate iron ions (Fe (II)). Depending on the pH value and the type of buffer-Fe (II) complex, its prooxidant activity and spatial distribution are influenced. The complex charge defines the interactions with the oil-water interface, i.e., positively charged interfaces repel positively charged complexes which keeps the prooxidant away. The mechanistic understanding of this work will help formulators and product developers to choose the right buffer and emulsifier combination for oxidation sensitive emulsions.