Abstract: An enzyme-linked Immunosorbent Assay (ELISA) was established for the detection of chloramphenicol in animal tissues. After ethyl acetate extraction and mixture of n-heane clean-up, a specific antibody(rabbit anti-CAP), enzyme labeled CAP (enzyme conjugate) and CAP standard or sample were added to the pre-coated wells followed by a single incubation step. The specific antibodies were bound by the immobillised antibodies and at the same time free CAP (present in the standard solution or sample) and enzyme conjugated CAP compete for the CAP antibody binding site (competitive enzyme immunoassay). After an incubation time of 1 hour, the non-bound (enzyme labelled) reagents were removed in a washing step. Bound enzyme conjugate transformed the colourless chromogen into a coloured product by the addition of a chromogen substrate (TMB). The substrate reaction was stopped by the addition of sulphuric acid. The colcour intensity was measured photometrically at 450 nm. The optical density was inversely proportional to the CAP concentration in the sample. As a result, the calibration curve should be virtually liner in the range of 0.025~2ng/ml. The limit of detection of the ELISA was 0.02 μg/kg. The rate of recovery was about 78.2%.