Abstract Toxicity investigations were conducted for four pyridinium chlorides belonging to cationic surface-active substances (CSAS), which differed from each other in the numbers of methyl groups (CH 3) in pyridinium ring. The crustacean Daphnia magna, the fish Lebistes reticulatus and the alga Scenedesmus quadricauda were chosen as biotests. Toxicity of examined preparations appeared to be very high but did not depend on their chemical structure. S. quadricauda was the most sensitive organism. Toxicity of intermediate products obtained in biological oxidation process was also examined. Biodegradation was conducted according to the “river water test”. It was found that only partial degradation took place while pyridinium chlorides constituted main energy and carbon source. Presence of biodegradation intermediate products was shown on the basis of 1H NMR analysis. Intermediates were not toxic to any biotests.