Determination of nifursol metabolites in poultry muscle and liver tissue. development and validation of a confirmatory method

• Published in: Analyst (London) 2005-01, Vol.130 (5), p.763-771
• Main Authors: MULDER, P. P. J, ZUIDEMA, T, KEESTRA, N. G. M, KOOIJ, P. J. F, ELBERS, I. J. W, VAN RHIJN, J. A
• Format: Article
• Language: English
• Subjects: 3-amino-2-oxazolidinone; Analytical chemistry; Animals; Antiprotozoal Agents - metabolism; Chemistry; Chromatographic methods and physical methods associated with chromatography; Chromatography, Liquid - methods; Drug Residues - analysis; Exact sciences and technology; Food Analysis - methods; Food Contamination - analysis; furazolidone metabolite; Liver - chemistry; Mass Spectrometry - methods; Muscles - chemistry; Nitrofurans - metabolism; Other chromatographic methods; performance liquid-chromatography; pigs; porcine tissues; Poultry - metabolism; residues; Spectrometric and optical methods; tandem mass-spectrometry
• ISSN: 0003-2654; 1364-5528
• DOI: 10.1039/b414320e

A method is described for the identification and quantitative determination of 3,5-dinitrosalicylic acid hydrazide (DSH), the marker residue of nifursol metabolites in poultry (turkey, broiler) muscle and liver tissue. The method is based on the acid-catalysed hydrolysis of tissue-bound metabolites to free DSH and in situ derivatisation with 2-nitrobenzaldehyde to the corresponding nitrophenyl derivative NPDSH. A structural analogue of DSH, 4-hydroxy-3,5-dinitrobenzoic acid hydrazide (HBH) was synthesised to serve as an internal standard. The analytes were isolated from the matrix by liquid-liquid extraction with ethyl acetate. Determination was performed by LC-MS/MS with negative electrospray ionisation. The [M - H](+) ions of NPDSH and NPHBH at m/z 374 were fragmented by collision induced dissociation (CID) producing transition ions at m/z 182, 183 and 226. The transition ions at m/z 182 and 226 were selected for monitoring of NPDSH while the transition ion at m/z 183 was selected for NPHBH. The method has been validated according to the EU criteria of Commission Decision 2002/657/EC at 0.5, 1.0 and 1.5 microg kg(-1) in muscle and liver tissue. A decision limit (CC(alpha)) was obtained of 0.04 and 0.025 microg kg(-1) in muscle and liver, respectively. Similarly a detection capability (CC(beta)) was obtained of 0.10 and 0.05 microg kg(-1) in muscle and liver, respectively. The introduction of HBH as an internal standard did not lead to a significant improvement of the quantitative performance of the method. In fact for liver better performance characteristics were obtained when the IS was not taken into account. Nevertheless, as a qualitative marker for recovery, HBH could still be very useful in the analysis of unknown samples.