title: Structure Elucidation, Relative LC-MS Response and In Vitro Toxicity of Azaspiracids 7-10 Isolated from Mussels (Mytilus edulis).
abstract: Azaspiracids (AZAs) are marine biotoxins produced by dinoflagellates that can accumulate in shellfish, which if consumed can lead to poisoning events. AZA7-10, 7-10, were isolated from shellfish and their structures, previously proposed on the basis of only LC-MS/MS data, were confirmed by NMR spectroscopy. Purified AZA4-6, 4-6, and 7-10 were accurately quantitated by qNMR and used to assay cytotoxicity with Jurkat T lymphocyte cells for the first time. LC-MS(MS) molar response studies performed using isocratic and gradient elution in both selected ion monitoring and selected reaction monitoring modes showed that responses for the analogues ranged from 0.3 to 1.2 relative to AZA1, 1. All AZA analogues tested were cytotoxic to Jurkat T lymphocyte cells in a time- and concentration-dependent manner; however, there were distinct differences in their EC50 values, with the potencies for each analogue being: AZA6 > AZA8 > AZA1 > AZA4 ≈ AZA9 > AZA5 ≈ AZA10. This data contributes to the understanding of the structure-activity relationships of AZAs.
authors.affiliation: †Marine Institute, Rinville, Oranmore, County Galway, Ireland.|‡School of Chemical and Pharmaceutical Sciences, Dublin Institute of Technology, Kevin Street, Dublin 8, Ireland.
authors.forename: Jane
authors.initials: J
authors.lastname: Kilcoyne
authors.affiliation: §School of Medicine, Wayne State University, Detroit, Michigan 48202, United States.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Twiner
authors.affiliation: ⊥Measurement Science and Standards, Biotoxin Metrology, National Research Council Canada, 1411 Oxford Street, Halifax, Nova Scotia B3H 3Z1, Canada.
authors.forename: Pearse
authors.initials: P
authors.lastname: McCarron
authors.affiliation: ⊥Measurement Science and Standards, Biotoxin Metrology, National Research Council Canada, 1411 Oxford Street, Halifax, Nova Scotia B3H 3Z1, Canada.
authors.forename: Sheila
authors.initials: S
authors.lastname: Crain
authors.affiliation: ⊥Measurement Science and Standards, Biotoxin Metrology, National Research Council Canada, 1411 Oxford Street, Halifax, Nova Scotia B3H 3Z1, Canada.
authors.forename: Sabrina D
authors.initials: SD
authors.lastname: Giddings
authors.affiliation: ‡School of Chemical and Pharmaceutical Sciences, Dublin Institute of Technology, Kevin Street, Dublin 8, Ireland.
authors.forename: Barry
authors.initials: B
authors.lastname: Foley
authors.affiliation: ∥Department of Chemistry, University of Oslo, N-0315 Oslo, Norway.
authors.forename: Frode
authors.initials: F
authors.lastname: Rise
authors.affiliation: ▽Ifremer, Laboratoire Phycotoxines, Rue de l'Ile d'Yeu, 44311 Nantes, France.
authors.forename: Philipp
authors.initials: P
authors.lastname: Hess
authors.affiliation: ○Norwegian Veterinary Institute, P.O. Box 750 Sentrum, 0106 Oslo Norway.
authors.forename: Alistair L
authors.initials: AL
authors.lastname: Wilkins
authors.affiliation: ○Norwegian Veterinary Institute, P.O. Box 750 Sentrum, 0106 Oslo Norway.|#Department of Pharmaceutical Chemistry, School of Pharmacy, University of Oslo, P.O. Box 1068 Blindern, N-0316 Oslo Norway.
authors.forename: Christopher O
authors.initials: CO
authors.lastname: Miles
mesh_terms: D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002853:Chromatography, Liquid; D006801:Humans; D019169:Jurkat Cells; D008387:Marine Toxins / Q000737:chemistry* / Q000633:toxicity*; D013058:Mass Spectrometry; D049880:Mytilus edulis / Q000737:chemistry*; D012758:Shellfish / Q000032:analysis*; D013141:Spiro Compounds / Q000737:chemistry* / Q000633:toxicity*; D013329:Structure-Activity Relationship; D013601:T-Lymphocytes / Q000166:cytology / Q000187:drug effects; D000095702:Polyether Toxins
chemical_list: D008387:Marine Toxins; D013141:Spiro Compounds; C406592:azaspiracid; D000095702:Polyether Toxins
keywords: Jurkat T; LC−MS molar response; NMR; azaspiracid; mass spectrometry; purification; structure confirmation; toxicity