Bioethanol production in North America has led to the production of considerable quantities of different co-products. Variation in nutrient profiles as well as nutrient availability among these co-products may lead to the formulation of imbalanced diets that may adversely affect animal performance. This study aimed to compare three types of dried distiller's grains with solubles [100% wheat DDGS (WDDGS); DDGS blend1 (BDDGS1, corn to wheat ratio 30:70); DDGS blend2 (BDDGS2, corn to wheat ratio 50:50)] and their different batches within DDGS type with regard to: (i) protein and carbohydrate sub-fractions based on Cornell Net Carbohydrate and Protein System (CNCPS); (ii) calculated energy values; and (iii) rumen degradation of dry matter (DDM), organic matter (DOM), crude protein (DCP), neutral detergent fibre (DNDF) and starch (Dstarch) at 36 and 72 h of ruminal incubations. Wheat DDGS had a lower intermediately (PB2, 136.4 vs. 187.4 g/kg DM) and a higher slowly degradable true protein (PB3, 142.2 vs.105.3 g/kg DM) than BDDGS1, but similar to those of BDDGS2. Sugar (CA4) was higher, whereas starch (PB1) and digestible fibre (PB3) were lower in WDDGS than in BDDGS1 and BDDGS2. All carbohydrate sub-fractions determined differed significantly between the two batches of BDDGS2. The BDDGS2 had the highest calculated energy values (TDN, DE(3×) , ME(3×) , NEL(3×) , NE(m) and NE(g) ) among the three DDGS types. The energy values were slightly different between the batches of the three DDGS types. At all incubation times, wheat DDGS had a significantly higher (p < 0.05) DDM, DOM, DCP and DNDF than both DDGS blends. Differences were observed between different batches within DDGS types with regard to in situ rumen degradation of DM, OM, CP, NDF and starch. In conclusion, differences were observed in protein and carbohydrate sub-fractions and in situ ruminal degradation of DM, OM, CP, NDF and starch among the three DDGS types and different batches within DDGS type. This indicates that the nutrients supplied to ruminants may not only differ among different types of DDGS but it may also differ among different batches within DDGS type.