A liquid chromatographic (HPLC) method has been developed to detect chloramphenicol (CAP) in fatty liver. After addition of water, samples were defatted with hexane partition and CAP was extracted by ethyl acetate. A solid-phase extraction protocol was used to recover CAP from the fatty liver extract. CAP was analyzed using a mobile phase of acetonitrile-0.005 M phosphate buffer, pH 7.9 (19:81), an ultraviolet detection wavelength of 278 nm and confirmation by scanning. The column used was a Nova-Pak C18(150 x 3.9 mm). The limit of detection was < 1 ng/g in fatty liver and the mean recovery of CAP was found to be 71% over the concentration range 5–20 ng/g. The analyte identity can be confirmed by the use of scanning detection and by LC/MS method for concentrations less than 10 ng/g.