title: Development, validation and application of an ultra high performance liquid chromatographic-tandem mass spectrometric method for the simultaneous detection and quantification of five different classes of veterinary antibiotics in swine manure.
abstract: In this study, a fast, simple and selective ultra high performance liquid chromatographic-tandem mass spectrometric (UHPLC-MS/MS) method for the simultaneous detection and quantification of colistin, sulfadiazine, trimethoprim, doxycycline, oxytetracycline and ceftiofur and for the detection of tylosin A in swine manure was developed and validated. First, a simple extraction procedure with acetonitrile and 6% trichloroacetic acid was carried out. Second, the supernatant was evaporated and the pellet was reconstituted in 1 ml of water/acetonitrile (80/20) and 0.1% formic acid. Extracts were filtered and analyzed by UHPLC-MS/MS on a Kinetex C18 column using gradient elution. The method developed was validated according to the criteria of Commission Decision 2002/657/EC. Recovery percentages varied between 94% and 106%, repeatability percentages were within the range of 1.7-9.2% and the intralaboratory reproducibility varied between 2.8% and 9.3% for all compounds, except for tylosin A for which more variation was observed resulting in a higher measurement uncertainty. The limit of detection and limit of quantification varied between 1.1 and 20.2 and between 3.5 and 67.3 μg/kg, respectively. This method was used to determine the presence and concentration of the seven antibiotic residues in swine manure sampled from ten different manure pits on farms where the selected antibiotics were used. A link was found between the antibiotics used and detected, except for ceftiofur which is injected at low doses and degraded readily in swine manure and was therefore not recovered in any of the samples. To the best of our knowledge, this is the first method available for the simultaneous extraction and quantification of colistin with other antibiotic classes. Additionally, colistin was never extracted from swine manure before. Another innovative aspect of this method is the simultaneous detection and quantification of five different classes of antibiotic residues in swine manure.
authors.affiliation: The Institute for Agricultural and Fisheries Research (ILVO), Technology and Food Science Unit, Brusselsesteenweg 370, 9090 Melle, Belgium; Ghent University, Department of Pathology, Bacteriology and Avian Diseases, Faculty of Veterinary Medicine, Salisburylaan 133, 9820 Merelbeke, Belgium. Electronic address: tina.vandenmeersche@ilvo.vlaanderen.be.
authors.forename: Tina
authors.initials: T
authors.lastname: Van den Meersche
authors.affiliation: The Institute for Agricultural and Fisheries Research (ILVO), Technology and Food Science Unit, Brusselsesteenweg 370, 9090 Melle, Belgium.
authors.forename: Els
authors.initials: E
authors.lastname: Van Pamel
authors.affiliation: The Institute for Agricultural and Fisheries Research (ILVO), Technology and Food Science Unit, Brusselsesteenweg 370, 9090 Melle, Belgium.
authors.forename: Christof
authors.initials: C
authors.lastname: Van Poucke
authors.affiliation: The Institute for Agricultural and Fisheries Research (ILVO), Technology and Food Science Unit, Brusselsesteenweg 370, 9090 Melle, Belgium.
authors.forename: Lieve
authors.initials: L
authors.lastname: Herman
authors.affiliation: The Institute for Agricultural and Fisheries Research (ILVO), Technology and Food Science Unit, Brusselsesteenweg 370, 9090 Melle, Belgium; Ghent University, Department of Pathology, Bacteriology and Avian Diseases, Faculty of Veterinary Medicine, Salisburylaan 133, 9820 Merelbeke, Belgium.
authors.forename: Marc
authors.initials: M
authors.lastname: Heyndrickx
authors.affiliation: The Institute for Agricultural and Fisheries Research (ILVO), Technology and Food Science Unit, Brusselsesteenweg 370, 9090 Melle, Belgium.
authors.forename: Geertrui
authors.initials: G
authors.lastname: Rasschaert
authors.affiliation: The Institute for Agricultural and Fisheries Research (ILVO), Technology and Food Science Unit, Brusselsesteenweg 370, 9090 Melle, Belgium.
authors.forename: Els
authors.initials: E
authors.lastname: Daeseleire
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000032:analysis*; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D008372:Manure / Q000032:analysis*; D015203:Reproducibility of Results; D013552:Swine; D053719:Tandem Mass Spectrometry
chemical_list: D000900:Anti-Bacterial Agents; D008372:Manure
keywords: Multi-residue; Swine manure; Ultra high performance liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS); Validation; Veterinary antibiotics