Scope Food matrix is generally believed to alter carotenoid bioavailability, but its effect on xanthophylls is usually limited. This study thus aims to decipher the digestion‐absorption process of lutein in the presence or not of a food matrix. Methods Lutein transfer to gastric‐like lipid droplets or artificial mixed micelles was assessed when lutein was added to test meals either as a pure molecule ((all‐ E )‐lutein) or in canned spinach (( Z ) + (all‐ E )‐lutein). The obtained mixed micelles were delivered to Caco‐2 cells to evaluate lutein uptake. Finally postprandial plasma lutein responses were compared in minipigs after the two test meals. Results Lutein transfer to gastric‐like lipid droplets and to mixed micelles was higher when lutein was added in spinach than when it was added as pure lutein (+614% and +147%, respectively, p < 0.05). Conversely, lutein uptake was less effective when micellar lutein was from a meal containing spinach than from a meal containing its pure form (−55%, p < 0.05). In minipigs, postprandial lutein response was delayed with spinach but not significantly different after the two test meals. Conclusion Opposite effects at the micellarization and intestinal cell uptake steps explain the lack of effect of spinach matrix on lutein bioavailability.