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

Abstract

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 sulphate. 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 minutes. 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, sulfadiazine, sulfamethazine, dinitolimide, ethopabate, carbadox and clopidol. The method is also capable of qualitatively analysing for 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.

Rapid multiresidue and multi-class screening for antibiotics and benzimidazoles in feed by ultra high performance liquid chromatography coupled to tandem mass spectrometry

“…Although many authors have used ACN or a mixture of ACN, methanol, and water (Cronly et al, 2010;Lopes et al, 2012;Wang et al, 2014) to precipitate proteins and eliminate other interferences, we got best results with methanol. In agreement with other studies (Borr as et al, 2013;Lopes et al, 2012), we found ACN to yield "cleaner" extracts, but methanol to have a much better extraction capacity: some compounds present after methanol extraction were undetectable after ACN extraction.…”

“…We did evaluate the utility of an additional clean-up step with an SPE or QuEChERS sorbent (we tested Florisil, C18, primary secondary amine, carbon nanotubes, a mixture of several), but as previously observed by Bosher et al 2010, the added step resulted in loss of certain compounds, so it was omitted. Other authors have likewise sought to avoid fastidious clean-up, such as Cronly et al (2010), who investigated the use of anhydrous sodium sulfate followed by defatting with hexane, and Boix et al (2014), Kaklamanos al. (2013), and N acher-Mestre et al (2013), who report screening methods for veterinary antimicrobials or organic pollutants in feed based on liquideliquid extraction with ACN.…”

“…After extraction of drug residues with organic solvents, various authors have used a purification step involving solid-phase extraction (SPE) (Aguilera-Luiz, Romero-Gonz alez, Plaza-Bolaños, Martinez Vidal, & Garrido Frenich, 2013;Ardsoongnearn, Boonbanlu, Kittijaruwattana, & Suntornsuk, 2014;Kantiani, Farr e, Grases, & Barcel o, 2010;Van Poucke, De Keyser, Baltusnikiene, McEvoy, & Van Peteghem, 2003;Vincent, Chedin, Yasar, & von Holst, 2008;Wang et al, 2014), liquideliquid clean-up (Cronly et al, 2010), filtration (Kaklamanos, Vincent, & Von Holst, 2013), or QuEChERS (Boscher, Guignard, Pellet, Hoffmann, & Bohn, 2010;Lopes et al, 2012). Others have used simpler means: dilution of the extract before analysis (Boix et al, 2014;Ch afer-Peric as, Maquieira, Puchades, Miralles, & Moreno, 2011) or protein precipitation by freezing the extract (N acher-Mestre, Ib añez, Serrano, P erez-S anchez, & Hern andez, 2013).…”

Scientific Opinion on Chloramphenicol in food and feed

“…The CCα values reported for these methods range from 0.01 to 0.15 µg/kg and the CCβ values range from 0.02 to 0.26 µg/kg. Cronly et al (2010b) describe an LC-MS/MS method for prohibited medicinal additives, including chloramphenicol, in pig and poultry compound feed, validated at a level of 100 µg/kg.…”

Development of an indirect competitive enzyme-linked immunosorbent assay for screening ethopabate residue in chicken muscle and liver

“…3 Residue monitoring is an effective way for assurance of the food safety and wholesomeness. Numerous methods, including spectrouorimetric assays, 2 spectrouorimetric methods, 4 gas chromatography, 5 high-performance liquid chromatography (HPLC) with UV detection 6 or by uorimetry, 7 liquid chromatography coupled to triple quadrupole mass spectrometry [8][9][10] or time-of-ight-mass spectrometry, 11 have been previously used for ETP residue analysis. Although these approaches are sensitive and reliable, these methods oen require extensive sample pretreatments and sophisticated laboratory equipment with highly trained operators.…”