Purification of nine sulfonamides from chicken tissues by immunoaffinity chromatography using two monoclonal antibodies

Abstract

An immunoaffinity chromatographic method was developed for the purification of 9 sulfonamides (sulfamethazine, sulfadimethoxine, sulfamerazine, sulfathiazole, sulfamethoxazole, suifamethizole, sulfadiazine, sulfamonomethoxine, and sulfapyridine) from chicken tissue (muscle and liver) samples. Two monoclonal antibodies (antisulfamethazine and antisulfamethoxazole) were simultaneously covalently coupled to CNBr-activated Sepharose 4B for the preparation of a re-usable immunoaffinity column. After extraction with methanol-water and purification by immunoaffinity column, the sulfonamides were determined by reversed-phase liquid chromatography and UV detection at 270 nm. The recoveries for each drug at fortification levels of 10-50 ng/g ranged from 74.1 to 108.9% with relative standard deviations from 1.9 to 11.5%. The limits of detection were 2 ng/g for each drug.