title: Determination of T-2 toxin, HT-2 toxin, and three other type A trichothecenes in layer feed by high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS)--comparison of two sample preparation methods.
abstract: A sensitive method for the simultaneous determination of T-2 toxin, HT-2 toxin, neosolaniol, T-2 triol, and T-2 tetraol in layer feed using high-performance liquid chromatography coupled to triple quadrupole mass spectrometry in the positive ionization mode (LC-ESI-MS/MS) is described. Two fast and easy clean-up methods-with BondElut Mycotoxin and MycoSep 227 columns, respectively-were tested. The separation of the toxins was conducted on a Pursuit XRs Ultra 2.8 HPLC column using 0.13 mM ammonium acetate as eluent A and methanol as eluent B. Detection of the mycotoxins was carried out in the multiple reaction monitoring (MRM) mode using ammonium adducts as precursor ions. Quantification of all analytes was performed with d3-T-2 toxin as an internal standard. The clean-up method with MycoSep 227 columns gave slightly better results for layer feed compared to the method using BondElut Mycotoxin columns (MycoSep 227: recovery between 50 and 63%, BondElut Mycotoxin: recovery between 32 and 67%) and was therefore chosen as the final method. The limits of detection ranged between 0.9 and 7.5 ng/g depending on the mycotoxin. The method was developed for the analysis of layer feed used at carry-over experiments with T-2 toxin in laying hens. For carry-over experiments, it is necessary that the method includes not only T-2 toxin but also the potential metabolites in animal tissues HT-2 toxin, neosolaniol, T-2 triol, and T-2 tetraol which could naturally occur in cereals used as feed stuff as well.
authors.affiliation: Institute of Animal Nutrition, Friedrich-Loeffler Institute (FLI), Federal Research Institute for Animal Health, Bundesallee 50, 38116, Braunschweig, Germany.|Institute of Food Chemistry, Westfälische Wilhelms-Universität Münster, Corrensstrasse 45, 48149, Münster, Germany.
authors.forename: Katrin
authors.initials: K
authors.lastname: Bernhardt
authors.affiliation: Institute of Animal Nutrition, Friedrich-Loeffler Institute (FLI), Federal Research Institute for Animal Health, Bundesallee 50, 38116, Braunschweig, Germany.
authors.forename: Hana
authors.initials: H
authors.lastname: Valenta
authors.affiliation: Institute of Animal Nutrition, Friedrich-Loeffler Institute (FLI), Federal Research Institute for Animal Health, Bundesallee 50, 38116, Braunschweig, Germany.
authors.forename: Susanne
authors.initials: S
authors.lastname: Kersten
authors.affiliation: Institute of Food Chemistry, Westfälische Wilhelms-Universität Münster, Corrensstrasse 45, 48149, Münster, Germany.
authors.forename: Hans-Ulrich
authors.initials: HU
authors.lastname: Humpf
authors.affiliation: Institute of Animal Nutrition, Friedrich-Loeffler Institute (FLI), Federal Research Institute for Animal Health, Bundesallee 50, 38116, Braunschweig, Germany. sven.daenicke@fli.bund.de.
authors.forename: Sven
authors.initials: S
authors.lastname: Dänicke
mesh_terms: D000821:Animal Feed / Q000032:analysis*; D000818:Animals; D002645:Chickens; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002523:Edible Grain / Q000737:chemistry; D057230:Limit of Detection; D009183:Mycotoxins / Q000032:analysis; D013605:T-2 Toxin / Q000031:analogs & derivatives* / Q000032:analysis*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014255:Trichothecenes / Q000737:chemistry*
chemical_list: D009183:Mycotoxins; D014255:Trichothecenes; D013605:T-2 Toxin; C012351:HT-2 toxin
keywords: LC-MS/MS; Layer feed; Metabolites; SPE clean-up; T-2 toxin; Trichothecenes