Alkaline phosphatase (ALP) conjugated to antibodies is often used in enzyme immunoassays (EIAs). These assays are notably sensitive to experimental conditions. A possible source of interference is bacterial ALP, which is released when bacterial contamination occurs in clinical analyzers. Preliminary experiments led to the selection of a detection kit, ALP source, and specific types of tubes for collecting water samples and performing assays. The release of ALP from various strains of bacteria identified in pure water was demonstrated (10-30×106 cfu/mL released 6-10 μU/μL). It was shown that ultrafiltration is totally efficient in removing ALP from water, while residual ALP activity (2.21 μU/μL after filtration of an ALP solution of 6.22 μU/μL) was observed after filtration using a 0.22-μm filter.