Bio-Methane Potential (BMP) tests are used to evaluate the suitability of a biomass for anaerobic digestion. BMP data are usually presented as the amount of methane produced from a kilogram of volatile solids (VS) or chemical oxygen demand (COD) of the substrate. However, the most used methods for determination of VS and COD are not always accurate. Oven drying may underestimate VS content due to loss of volatile organic compounds, and incomplete chemical oxidation may lead to underestimation of COD content. Bomb calorimetry is an attractive alternative to COD measurements, because the physical state of the biomass sample does not influence the measurement, and because sample preparation is limited. In this study, 11 biomass samples, wet and dry, were analyzed with different methods for organic content determination. COD (determined by bomb calorimetry and by wet chemistry) and VS (by Karl Fischer titration and loss on drying; LOD) were compared, and used for determination of BMP. In general, the BMP estimated on a VS basis were higher than those estimated on COD basis. For certain biomass samples the method for VS determination also greatly influenced the results; for fishery waste the BMP was estimated as 928 L kg(-1) based on LOD-VS compared to 394 L kg(-1) based on KF-LOD. Thus, this study shows that determination of organic content is not trivial and the method of choice strongly influences the estimation of bio-methane potentials. Bomb calorimetry offers a possibility to measure energy content directly, independent of biomass composition and physical state. (C) 2017 Elsevier Ltd. All rights reserved.