title: A validated LC-MS/MS method for thyroid hormone determination in sea lamprey (Petromyzon marinus) plasma, gill, kidney and liver.
abstract: A UPLC-MS/MS method was developed to provide a reproducible, sensitive and quantitative assay to determine thyroid hormones in sea lamprey tissues and plasma. l-Thyroxine (T4) and its two triiodo-thyronine isomers have been simultaneously quantified and validated for plasma, gill, liver, and kidney matrices. Multiple sample preparation techniques were investigated to achieve optimal sample matrix digestion and clean-up. Enzymatic digestion followed by protein precipitation was selected to process the samples. The developed method exhibited excellent linearity for all analytes with regression coefficients higher than 0.99 for concentrations ranged from 10 to 50,000pg/mL. The limit of detection (LOD) was under 1pg/mL while the limit of quantification (LOQ) was estimated as 10pg/mL. This method was validated according to the FDA guidance and applied to determine thyroid hormone levels in plasma, gill and kidney of sea lamprey exposed to a sex pheromone. With appropriate implementation and further validation, this method could be applied to tissues in other species including humans.
authors.affiliation: Department of Fisheries and Wildlife, Michigan State University, Room 13 Natural Resources Building, 480 Wilson Road, East Lansing, MI 48824, USA. Electronic address: ugo.bussy@gmail.com.
authors.forename: Ugo
authors.initials: U
authors.lastname: Bussy
authors.affiliation: Department of Fisheries and Wildlife, Michigan State University, Room 13 Natural Resources Building, 480 Wilson Road, East Lansing, MI 48824, USA.
authors.forename: Yu-Wen
authors.initials: YW
authors.lastname: Chung-Davidson
authors.affiliation: Department of Fisheries and Wildlife, Michigan State University, Room 13 Natural Resources Building, 480 Wilson Road, East Lansing, MI 48824, USA.
authors.forename: Ke
authors.initials: K
authors.lastname: Li
authors.affiliation: Department of Fisheries and Wildlife, Michigan State University, Room 13 Natural Resources Building, 480 Wilson Road, East Lansing, MI 48824, USA.
authors.forename: Skye D
authors.initials: SD
authors.lastname: Fissette
authors.affiliation: Department of Fisheries and Wildlife, Michigan State University, Room 13 Natural Resources Building, 480 Wilson Road, East Lansing, MI 48824, USA.
authors.forename: Ethan G
authors.initials: EG
authors.lastname: Buchinger
authors.affiliation: Department of Fisheries and Wildlife, Michigan State University, Room 13 Natural Resources Building, 480 Wilson Road, East Lansing, MI 48824, USA. Electronic address: liweim@msu.edu.
authors.forename: Weiming
authors.initials: W
authors.lastname: Li
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods*; D005880:Gills / Q000737:chemistry; D007668:Kidney / Q000737:chemistry; D057230:Limit of Detection; D016014:Linear Models; D008099:Liver / Q000737:chemistry; D048428:Petromyzon / Q000378:metabolism*; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013963:Thyroid Hormones / Q000032:analysis* / Q000378:metabolism; D014018:Tissue Distribution
chemical_list: D013963:Thyroid Hormones
keywords: Sea lamprey; T3; T4; Thyroid hormones; UPLC–MS/MS; rT3