Determination of rodenticides and related metabolites in rabbit liver and biological matrices by liquid chromatography coupled to Orbitrap high resolution mass spectrometry.
An analytical method based on ultra-high performance liquid chromatography (UHPLC) coupled to Orbitrap high resolution mass spectrometry was developed for the determination of rodenticides (bromadiolone, brodifacoum, difenacoum, chlorophacinone, diphacinone, coumachlor and warfarin) in liver matrix. Different extraction conditions were tested, obtaining the best results when the "dilute and shoot" method (acidified acetonitrile as extraction solvent) and a clean-up step with primary secondary amine (PSA) were used. The optimized method was validated, obtaining recoveries ranging from 60 to 120%. Repeatability and reproducibility were evaluated obtaining values lower than 20%, except for brodifacoum at 10μg/kg. Limits of quantification (LOQs) ranged from 0.1 to 0.5μg/kg, except for brodifacoum, which was 100μg/kg. Six liver samples were analyzed and diphacinone and chlorophacinone were detected in three samples at concentrations ranging from 4μg/kg to 13μg/kg. Moreover a retrospective screening of rodenticide metabolites in those samples and in animal forensic samples was developed based on Orbitrap capabilities. Brodifacoum was detected in three samples, and warfarin alcohol, which is a metabolite of warfarin, was also detected in one sample.
Research Group "Analytical Chemistry of Contaminants", Department of Chemistry and Physics, Research Centre for Agricultural and Food Biotechnology (BITAL), University of Almeria, Agrifood Campus of International Excellence, ceiA3, E-04120 Almeria, Spain.
Marina
M
López-García
Research Group "Analytical Chemistry of Contaminants", Department of Chemistry and Physics, Research Centre for Agricultural and Food Biotechnology (BITAL), University of Almeria, Agrifood Campus of International Excellence, ceiA3, E-04120 Almeria, Spain.
Roberto
R
Romero-González
Research Group "Analytical Chemistry of Contaminants", Department of Chemistry and Physics, Research Centre for Agricultural and Food Biotechnology (BITAL), University of Almeria, Agrifood Campus of International Excellence, ceiA3, E-04120 Almeria, Spain. Electronic address: agarrido@ual.es.
Antonia Garrido
AG
Frenich
D015110:4-Hydroxycoumarins / Q000737:chemistry; D000097:Acetonitriles / Q000737:chemistry; D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods; D003374:Coumarins / Q000737:chemistry; D007189:Indans / Q000737:chemistry; D008099:Liver / Q000737:chemistry* / Q000378:metabolism*; D013058:Mass Spectrometry / Q000379:methods; D010630:Phenindione / Q000031:analogs & derivatives / Q000737:chemistry; D011817:Rabbits; D015203:Reproducibility of Results; D012189:Retrospective Studies; D012378:Rodenticides / Q000737:chemistry* / Q000378:metabolism*; D014859:Warfarin / Q000737:chemistry
D015110:4-Hydroxycoumarins; D000097:Acetonitriles; D003374:Coumarins; D007189:Indans; D012378:Rodenticides; C004662:chlorophacinone; C100262:diphenadione; D010630:Phenindione; D014859:Warfarin; C013418:bromfenacoum; C021636:bromadiolone; C012903:difenacoum; C412550:coumachlor; C032159:acetonitrile
High resolution mass spectrometry; Liver matrix; Metabolites; Orbitrap; Retrospective analysis; Rodenticides