Multiresidue analysis of cephalosporin antibiotics in bovine milk based on molecularly imprinted polymer extraction followed by liquid chromatography-tandem mass spectrometry.
This work reports the preparation of molecularly imprinted polymers (MIPs) selective to cephalosporin (CF) antibiotics, and their application as molecularly imprinted solid-phase extraction (MISPE) sorbents for the determination of these antimicrobials in milk samples. Several functional monomers and cross-linkers have been screened to select the best combination that provides high selectivity for the simultaneous multiresidue extraction of cefthiofur (THIO), cefazolin (AZO), cefquinome (QUI), cephapirin (API), cephalexin (ALE) and cephalonium (ALO) from the samples. The novel MIPs were prepared by a non-covalent imprinting approach in the form of spherical microparticles using the synthetic surrogate molecule sodium 7-(2-biphenylylcarboxamido)-3-methyl-3-cepheme-4-carboxylate, N-3,5-bis(trifluoromethyl)phenyl-N'-4-vinylphenyl urea (VPU) as functional monomer, and divinylbenzene (DVB) as crosslinking agent in a 1:2:20 molar ratio. The optimized MISPE method allowed the extraction of the target antimicrobials from raw cow milk samples using a selective washing with 5mL methanol/2-[4-(2-hydroxyethyl)-1-piperazinyl]ethanesulfonic acid (HEPES) buffer (0.1M, pH 7.5) (2:98, v/v) to remove the non-specifically retained compounds, followed by elution with 1mL of trifluoroacetic acid (TFA) in methanol (0.1:99.9, v/v). The extracts have been analysed by UHPLC-MS/MS and the analytical method has been validated according to EU guideline 2002/657/EC. The limits of quantification (S/N=10) were in the 1.7-12.5μgkg-1range, well below the maximum residue limits (MRLs) currently established for the quantified cephalosporins in milk samples. The developed MIP allows mutiresidual determination of the six cephalosporin antibiotics mentioned above, significantly broadening the application to food analysis of MISPE methods.
Department of Analytical Chemistry, Faculty of Chemistry, Universidad Complutense de Madrid, E-28040 Madrid, Spain; Institute of Materials Research and Engineering (IMRE), University of Havana, Havana 10400, Cuba.
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Baeza
Department of Analytical Chemistry, Faculty of Chemistry, Universidad Complutense de Madrid, E-28040 Madrid, Spain; CEI Campus Moncloa, UCM-UPM, Avda Complutense s/n, E-28040 Madrid, Spain. Electronic address: jurracar@quim.ucm.es.
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Urraca
Department of Organic Chemistry, Faculty of Chemistry, Universidad Complutense de Madrid, E-28040 Madrid, Spain.
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Chamorro
Department of Organic Chemistry, Faculty of Chemistry, Universidad Complutense de Madrid, E-28040 Madrid, Spain.
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Orellana
Institute for Food and Agricultural Research and Technology, Food Safety Program, Granja Camps i Armet s/n, Monells 17121, Girona, Spain.
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Castellari
Department of Analytical Chemistry, Faculty of Chemistry, Universidad Complutense de Madrid, E-28040 Madrid, Spain. Electronic address: mcmbondi@quim.ucm.es.
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Moreno-Bondi
D000818:Animals; D000900:Anti-Bacterial Agents / Q000032:analysis*; D002417:Cattle; D002511:Cephalosporins / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D003432:Cross-Linking Reagents; D004350:Drug Residues / Q000032:analysis*; D006531:HEPES; D000432:Methanol; D008855:Microscopy, Electron, Scanning; D008892:Milk / Q000737:chemistry*; D054802:Molecular Imprinting; D011108:Polymers; D012997:Solvents; D053719:Tandem Mass Spectrometry; D014269:Trifluoroacetic Acid
D000900:Anti-Bacterial Agents; D002511:Cephalosporins; D003432:Cross-Linking Reagents; D011108:Polymers; D012997:Solvents; D014269:Trifluoroacetic Acid; D006531:HEPES; D000432:Methanol
Cephalosporins; MISPE; Milk analysis; Molecularly imprinted polymers