The efficiencies of denitrification and enhanced biological phosphorus removal (EBPR) in biological nutrient removal (BNR) activated sludge systems are strongly dependent on the availability of appropriate carbon sources. Due to high costs of commercial compounds (such as methanol, ethanol, acetic acid etc.) and acclimation periods (usually) required, the effective use of internal carbon sources for denitrification is preferred. The aim of this study was to determine the immediate effects of slowly biodegradable substrates on the denitrification capability and phosphate release/uptake interactions for a full-scale biomass process from the "Wschod" wastewater treatment plant (WWTP) in Gdansk (Poland). Since it is hard to distinguish the slowly biodegradable substrate in a direct way, a novel procedure based on batch experiments was developed and implemented. The laboratory experiments were carried out in two parallel, fully automatic batch reactors with the settled wastewater without pretreatment and after coagulation-flocculation. The removal of colloidal and particulate fractions resulted in the reduced observed process rates, such as denitrification, phosphate release and phosphate uptake (under aerobic and anoxic conditions). The reduction ranged from approximately 14% for the anaerobic P release to approximately 46% for the anoxic P uptake.