Extra virgin olive oil (EVOO) and olives, key sources of unsaturated fatty acids in the Mediterranean diet, provide health benefits to humans. Nitric oxide (•NO) and nitrite (NO₂⁻)-dependent reactions of unsaturated fatty acids yield electrophilic nitroalkene derivatives (NO₂-FA) that manifest salutary pleiotropic cell signaling responses in mammals. Herein, the endogenous presence of NO₂-FA in both EVOO and fresh olives was demonstrated by mass spectrometry. The electrophilic nature of these species was affirmed by the detection of significant levels of protein cysteine adducts of nitro-oleic acid (NO₂-OA-cysteine) in fresh olives, especially in the peel. Further nitration of EVOO by NO₂⁻ under acidic gastric digestive conditions revealed that human consumption of olive lipids will produce additional nitro-conjugated linoleic acid (NO₂-cLA) and nitro-oleic acid (NO₂-OA). The presence of free and protein-adducted NO₂-FA in both mammalian and plant lipids further affirm a role for these species as signaling mediators. Since NO₂-FA instigate adaptive anti-inflammatory gene expression and metabolic responses, these redox-derived metabolites may contribute to the cardiovascular benefits associated with the Mediterranean diet.