Biodegradation is the most important attenuation process for most of organic chemicals in the environment. This process decides whether the organic substance itself or its degradation products rests in the environment and should be considered for a further risk assessment. This work presents the development of a water sediment screening test, based on OECD guideline 308, with a high significance to environmental conditions and with a good reproducibility and consistency of results. The increased reproducibility was achieved by creating an artificial and standardized medium, based on the existing OECD guidelines OECD 302C, 301D and 218. Each test consisted of five different series: blank, quality control, test, toxicity control and abiotic control. Biodegradation was assessed by measurement of pressure difference in closed vessels using the OxiTop(®) system. Aniline, diethylene glycol and sodium acetate were used to optimize and validate test conditions. Additionally, two pharmaceuticals: Acetaminophen and ciprofloxacin (CIP) were tested as an example of possible test application. Acetaminophen was mainly removed from the system by biodegradation whereas CIP was removed from water phase by sorption onto sediment. Water sediment test proved to be a promising tool for the biodegradation investigation of chemicals in the water-sediment interface.