Laboratory batch experiments were conducted at 20 degrees C to investigate the potential of primary sludge fermentation for the generation of readily biodegradable substrate and to evaluate the effect of fermentation products on mass balance for organic carbon, nitrogen and phosphorus, emphasizing COD fractionation. Fermentation converted between 18 to 30% of the initial volatile suspended solids in the sludge into soluble biodegradable COD. The volatile fatty fraction of the soluble COD was approximately 85% after the fermentation process. The average volatile fatty acid composition in fermentation involved 50% acetic acid, 33% propionic acid, 9% butyric acid and 8% valeric acid, indicating that the most important volatile fatty acid obtained during the biological fermentation process was acetate with more than half of total VFA concentration, which is one of the most important carbon sources for denitrification and biological nutrient removal processes. The recoverable fraction of the fermented sludge supernatant could potentially increase the readily biodegradable COD content of the primary effluent by 5%, together with a potential increase of the soluble nitrogen and phosphorus content by 2%.