Development and validation of a rapid multi-class method for the confirmation of fourteen prohibited medicinal additives in pig and poultry compound feed by liquid chromatography–tandem mass spectrometry

• Published in: Journal of pharmaceutical and biomedical analysis 2010-12, Vol.53 (4), p.929-938
• Main Authors: Cronly, Mark, Behan, P., Foley, B., Malone, E., Earley, S., Gallagher, M., Shearan, P., Regan, L.
• Format: Article
• Language: English
• Subjects: Acetonitrile; Analysis; Analytical, structural and metabolic biochemistry; Animal Feed - analysis; Animals; Antibiotics; Biological and medical sciences; Chromatography, Liquid - methods; Fundamental and applied biological sciences. Psychology; General pharmacology; LC–MS/MS; Medical sciences; Pharmaceutical Preparations - analysis; Pharmacology. Drug treatments; Pig and poultry compound feed; Poultry; Reproducibility of Results; Swine; Tandem Mass Spectrometry - methods; Validation
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/j.jpba.2010.06.027

A confirmatory method has been developed to allow for the analysis of fourteen prohibited medicinal additives in pig and poultry compound feed. These compounds are prohibited for use as feed additives although some are still authorised for use in medicated feed. Feed samples are extracted by acetonitrile with addition of sodium sulfate. The extracts undergo a hexane wash to aid with sample purification. The extracts are then evaporated to dryness and reconstituted in initial mobile phase. The samples undergo an ultracentrifugation step prior to injection onto the LC–MS/MS system and are analysed in a run time of 26 min. The LC–MS/MS system is run in MRM mode with both positive and negative electrospray ionisation. The method was validated over three days and is capable of quantitatively analysing for metronidazole, dimetridazole, ronidazole, ipronidazole, chloramphenicol, sulfamethazine, dinitolimide, ethopabate, carbadox and clopidol. The method is also capable of qualitatively analysing for sulfadiazine, tylosin, virginiamycin and avilamycin. A level of 100 μg kg −1 was used for validation purposes and the method is capable of analysing to this level for all the compounds. Validation criteria of trueness, precision, repeatability and reproducibility along with measurement uncertainty are calculated for all analytes.