title: Determination of okadaic acid, dinophysistoxin-1 and related esters in Greek mussels using HPLC with fluorometric detection, LC-MS/MS and mouse bioassay.
abstract: An approach involving both chemical and biological methods was undertaken for the detection and quantification of the marine toxins okadaic acid (OA), dinophysistoxin-1 (DTX-1) and their respective esters in mussels from different sampling sites in Greece during the period 2006-2007. Samples were analyzed by means of a) high performance liquid chromatography with fluorometric detection (HPLC-FLD), using 9-athryldiazomethane (ADAM), as a pre-column derivatization reagent, b) liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) and c) the mouse bioassay. Free OA and DTX-1 were determined by both HPLC-FLD and LC-MS/MS, while their respective esters were determined only by LC-MS/MS after alkaline hydrolysis of the samples. The detection limit (L.O.D.) and quantification limit (L.O.Q.) of the HPLC-FLD method were 0.015 microg/g HP and 0.050 microg/g HP, respectively, for OA. The detection limit (L.O.D.) and quantification limit (L.O.Q.) of the LC-MS/MS method were 0.045 microg/g HP and 0.135 microg/g HP, respectively, for OA. Comparison of results between the two analytical methods showed excellent agreement (100%), while both HPLC-FLD and LC-MS/MS methods showed an agreement of 97.1% compared to the mouse bioassay.
authors.affiliation: Laboratory of Food Chemistry, Department of Chemistry, University of Ioannina, 45110 Ioannina, Greece.
authors.forename: Artemis P
authors.initials: AP
authors.lastname: Louppis
authors.forename: Anastasia V
authors.initials: AV
authors.lastname: Badeka
authors.forename: Panagiota
authors.initials: P
authors.lastname: Katikou
authors.forename: Evangelos K
authors.initials: EK
authors.lastname: Paleologos
authors.forename: Michael G
authors.initials: MG
authors.lastname: Kontominas
mesh_terms: D000818:Animals; D001681:Biological Assay; D049872:Bivalvia / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006115:Greece; D057230:Limit of Detection; D051379:Mice; D019319:Okadaic Acid / Q000032:analysis* / Q000633:toxicity / Q000031:analogs & derivatives; D011714:Pyrans / Q000032:analysis* / Q000633:toxicity; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D011714:Pyrans; D019319:Okadaic Acid; C051904:dinophysistoxin 1