AUTOMATED HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC DETERMINATION OF CHLORAMPHENICOL IN MILK AND SWINE MUSCLE-TISSUE USING ONLINE IMMUNOAFFINITY SAMPLE CLEANUP

An on-line high-performance liquid immunoaffinity chromatographic (HPLIAC) system for the direct determination of chloramphenicol in milk and swine muscle tissue is described. The system consisted of a dual-column system in which an HPLIAC column was directly coupled to an RP-8 high-performance liquid chromatographic column. Skimmed and deproteinated milk or aqueous muscle tissue extract was directly injected into the HPLIAC column. After a washing step with phosphate-buffered saline, chloramphenicol was desorbed by a glycine-NaCl buffer (pH 2.8) and directly concentrated on the RP-8 column. Next, chromatography was carried out using acetonitrile-sodium acetate buffer as the mobile phase. Chloramphenicol was detected at 280 nm. Mean recoveries from spiked raw milk were 70 +/- 2% (1-50 mug/kg) and from spiked swine muscle tissue 64 +/- 2% (10-50 mug/kg). The calibration curves were linear in the range 1-200 mug/kg spiking levels. Limits of determination were 1 mug/kg for milk and 10 mug/kg for muscle tissue.