Omega-3 fatty acids such as eicosapentaenoic acid (EPA)and docosahexaenoicacid (DHA) are essential for human health but prone to oxidation.While esterification location is known to influence the stabilityof omega-3 in triacylglycerols (TAGs) in oxidation trials, their oxidativebehavior in the gastrointestinal tract is unknown. Synthesized ABA-and AAB-type TAGs containing DHA and EPA were submitted to static in vitro digestion for the first time. Tridocosahexaenoinand DHA as ethyl esters were similarly digested. Digesta were analyzedby gas chromatography, liquid chromatography-mass spectrometry,and nuclear magnetic resonance spectroscopy. Besides the formationof di- and monoacylglycerols, degradation of hydroperoxides was detectedin ABA- and AAB-type TAGs, whereas oxygenated species increased intridocosahexaenoin. Ethyl esters were mainly unaffected. EPA was expectedlyless susceptible to oxidation prior to and during the digestion process,particularly in sn-2. These results are relevantfor the production of tailored omega-3 structures to be used as supplementsor ingredients.