This study was designed to investigate whether dietary neutral detergent fiber (NDF):starch ratio could be considered as a nutritional indicator to evaluate carbohydrate composition and manipulate milk production and composition synthesis. Eight primiparous dairy cows were assigned to four total mixed rations with NDF:starch ratios of 0.86, 1.18, 1.63 and 2.34 from T1 to T4 in a replicated 4×4 Latin square design. Dry matter intake and milk production were decreased from T1 to T4. Digestibility of dry matter, organic matter, NDF and crude protein were linearly decreased from T1 to T4. As NDF:starch ratio increased, milk protein content and production, and milk lactose content and production were linearly reduced. However, milk fat content was linearly increased from T1 to T4. Quadratic effect was observed on milk fat production with the highest level in T3. Averaged rumen pH was linearly increased from T1 to T4, and subacute rumen acidosis occurred in T1. Ruminal propionate and butyrate concentration were linearly decreased, and microbial crude protein and metabolizable protein decreased from T1 to T4. It is concluded that NDF:starch ratio can be considered as a potential indicator to evaluate dietary carbohydrate composition and manipulate milk production and composition synthesis.