A sensitive streptavidin‐biotin enzyme‐linked immunosorbent assay (ELISA) for the detection of chloramphenicol (CAP) in milk is described. This test is based on a procedure developed earlier for the direct detection of CAP in crude aqueous meat extracts. Milk samples were defatted by centrifugation, filtered and directly submitted to the ELISA procedure. The results were compared with the values obtained by analysis of a part of the sample from which CAP was removed by an immobilized monoclonal antibody preparation. In spiked samples the presence of CAP at concentrations of 1·0 μg/kg and higher can be easily demonstrated. The possibility of the interpretation of the ELISA results with a statistical method, i.e. the so‐called standard deviation ratio (SDR) procedure, was also investigated.