We determined apparent total-tract fatty acid (FA) digestibility of lactating dairy cows fed palmitic acid (C16:0)-enriched supplements differing in their degree of esterification and FA profile. Our analysis utilized individual observations (n = 385) of mid-lactation Holstein dairy cows from 7 Latin square design studies. Diets (% DM) contained (mean ± SD) 30.6 ± 3.66 NDF, 27.1 ± 2.16 starch, and 4.0 ± 0.97 total FA. C16:0 supplements were classified based on the degree of esterification and FA profile as follows: a) C16:0_FFA (prilled free FA supplements containing ~85% C16:0 and ~6.0% C18:1); b) C16:0_TAG (prilled triglyceride supplements containing ~80% C16:0 and ~13% C18:1); and c) C16:0_blend (blend of free FA and Ca-salt supplements containing ~80% C16:0 and ~10% C18:1). Data were analyzed using a mixed model including treatment as a fixed effect and study, cow (study), and period (study) as random effects. To estimate supplement FA digestibility, we used a Lucas test with a regression of the intake of supplemental FA (g/d) on supplemental absorbed FA (g/d); slopes indicate true digestibility and intercepts endogenous synthesis. Apparent total-tract FA digestibility (FAd; %) for control diets (no supplemental fat) was 76.1 ± 0.63 (mean ± SEM) and was not different across studies (P = 0.85). We did not observe differences in FAd between control and C16:0_blend supplemented diets (76.1 ± 0.63 vs. 76.3 ± 0.89, P = 0.92). In contrast, compared with control, C16:0_FFA (76.1 ± 0.63 vs. 71.9 ± 0.66; P < 0.01) and C16:0_TAG (76.1 ± 0.63 vs. 69.8 ± 0.77, P < 0.01) supplemented diets decreased FAd. C16:0_blend supplemented diets increased FAd compared with C16:0_FFA (P < 0.01) and C16:0_TAG (P < 0.01) supplemented diets. Additionally, C16:0_FFA supplemented diets increased FAd compared with C16:0_TAG (P < 0.05) supplemented diets. Digestibility for supplemental fats estimated by Lucas test were 69.2 ± 1.26, 60.6 ± 1.47, and 76.4 ± 1.15 for C16:0_FFA, C16:0_TAG, and C16:0_blend, respectively. Our results demonstrate that the degree of esterification and FA profile of C16:0-enriched supplements impact total FA digestibility.