Use of solid phase extraction for the isolation and clean-up of a derivatised furazolidone metabolite from animal tissues

Abstract

A method is presented for the determination of protein-bound residues of furazolidone in animal tissue. The use of furazolidone in food-producing animals has been banned in the EU. Illegal use of furazolidone can be monitored most effectively by testing for bound residues containing the 3-amino-2-oxazolidone (AOZ) moiety which, unlike the parent drug, is stable and can be detected for prolonged periods after cessation of treatment. This paper reports the development of an extraction and clean-up procedure for AOZ from liver using solid phase extraction. The method replaces solvent extraction and provides extensive sample clean-up with removal of approximately 99% of the derivatising agent, 2-nitrobenzaldehyde, which may interfere with the determination. It also offers the advantage of being suitable for automation, thereby increasing throughput of samples. The extraction procedure may be used for HPLC and ELISA screening techniques. The method has been validated in fortified and incurred pig liver samples, yielding mean recovery of AOZ in excess of 60%.