Tissue-bound residues of thiabendazole (TBZ), a veterinary anthelmintic and postharvest fungicide, are formed when this compound is incubated with rabbit hepatocytes or administered to mice or pigs. Several pretreatment steps were investigated for removing free TBZ and metabolites prior to the release of bound residues, and three procedures were evaluated for the release of bound residues from solvent-extracted rabbit hepatocytes: incubation under acidic conditions, enzymatic action using cystathionine beta-lyase, and Raney nickel desulfurization. Immunoaffinity chromatography utilizes monoclonal antibodies capable of binding TBZ or its 5-hydroxy metabolite enabled isolation of crossreactive residue fractions. Residues released from incurred pig liver and isolated by immunoaffinity included TBZ, as determined by HPLC with photodiode array detection. The methodology described should facilitate food safety assessments of TBZ.