We aimed to evaluate the effects of feeding sugarcane liquid molasses (LM) with or without a commercial buffer mix (BFM) on ruminal fermentation parameters, milk fatty acid (FA) profile, and milk yield and composition in dairy cows fed high-concentrate diets (35:65 forage-to-concentrate ratio). Eight multiparous Holstein cows (4 ruminally cannulated) averaging 165 +/- 12 d in milk at the beginning of the study were randomly assigned to a replicated 4 x 4 Latin square design with a 2 x 2 factorial arrangement of treatments. Each period lasted 21 d with 14 d for diet adaptation and 7 d for data and sample collection. Cows were fed the following diets: (1) no LM or BFM supplementation (CTRL), (2) LM without BFM supplementation (MOL), (3) BFM without LM supplementation (BUF), and (4) LM plus BFM supplementation (COMBO). These 4 isonitrogenous and isoenergetic diets were formulated by replacing (dry matter basis) 5\% ground corn with LM, whereas BFM replaced wheat bran at 0.8\% of the diet. Significant LM x BFM interactions were observed for the duration of ruminal pH below 5.8, molar proportion of propionate, acetate-to-propionate ratio, milk proportions of trans-10 18:1 and total trans FA, and concentration and yield of milk fat. Feeding MOL and BUF alone were effective on reducing the time that ruminal pH remained below 5.8 compared with the CTRL treatment, and the COMBO diet decreased it further. A similar pattern was observed for the ruminal molar proportion of propionate. The milk proportions of trans-10 18:1 and total trans FA dropped significantly with BFM or LM supplementation versus cows fed CTRL, and the COMBO diet decreased these variables further. Note, however, that these changes elicited by the COMBO diet were not in the same magnitude as those caused by MOL or BUF fed alone. The ruminal molar proportion of acetate increased with the BUF diet and that of butyrate increased in cows fed MOL, but mean ruminal pH was not affected by treatments. Diets with LM resulted in increased concentrations of short- and medium-chain FA in milk fat. The yield of 3.5\% fat-corrected milk increased significantly in cows fed MOL or BUF due to the improved concentration of milk fat. A trend and a significant increase for energycorrected milk were observed with feeding MOL or BUF, respectively. Overall, inclusion of LM and BFM appears to reduce milk trans-10 18:1 FA and total trans FA by modulating ruminal pH and volatile FA profile in cows fed high-concentrate diets.