Lipid oxidation is the main reason for the limited shelf life of mayonnaise. One of the main catalysts of this process is iron, which is introduced in its ferric (Fe(III)) form via phosvitin, an egg yolk phosphoprotein rich in phosphoserines. The binding of Fe(III) to phosvitin and its ability to establish a redox couple with Fe(II) is believed to determine the oxidation rate of unsaturated lipids. In this work, a 31P NMR based method was developed to quantify loading of phosvitin with Fe(III) and its reductive release. Both features could be quantified in model phosvitin solutions by exploiting the paramagnetic broadening of 31P NMR signal of phosphoserine residues by Fe(III). This method was then successfully applied to quantify the phosvitin‐Fe(III) loading in mayonnaise water phase by liquid NMR, whereas 31P NMR MAS could only provide a qualitative measure. The 31P NMR method showed a direct relation between loading of the Fe(III)‐phosvitin complex and lipid oxidation.