title: Biomarkers of neurotoxic shellfish poisoning.
abstract: Urine specimens from patients diagnosed with neurotoxic shellfish poisoning (NSP) were examined for biomarkers of brevetoxin intoxication. Brevetoxins were concentrated from urine by using solid-phase extraction (SPE), and analyzed by enzyme-linked immunosorbent assay (ELISA) and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Urine extracts were fractionated by LC, and fractions analyzed for brevetoxins by ELISA. In subsequent LC-MS/MS analyses, several brevetoxin metabolites of B-type backbone were identified, with elution profiles consistent with those of ELISA. The more abundant brevetoxin metabolites in urine were characterized structurally by LC-MS/MS. With the exception of BTX-3, brevetoxin metabolites in urine differed from those found in shellfish and in shellfish meal remnants. Proposed structures of these major urinary metabolites are methylsulfoxy BTX-3, 27-epoxy BTX-3, and reduced BTX-B5. BTX-3 was found in all specimens examined. BTX-3 concentrations in urine, as determined by LC-MS/MS, correlated well with composite toxin measurements by ELISA (r(2)=0.96). BTX-3 is a useful biomarker for confirmation of clinical diagnosis of NSP.
authors.affiliation: Gulf Coast Seafood Laboratory, U.S. Food and Drug Administration, P.O. Box 158, 1 Iberville Drive, Dauphin Island, AL 36528-0158, USA. ann.abraham@fda.hhs.gov
authors.forename: Ann
authors.initials: A
authors.lastname: Abraham
authors.forename: Steven M
authors.initials: SM
authors.lastname: Plakas
authors.forename: Leanne J
authors.initials: LJ
authors.lastname: Flewelling
authors.forename: Kathleen R
authors.initials: KR
authors.lastname: El Said
authors.forename: Edward L E
authors.initials: EL
authors.lastname: Jester
authors.forename: Hudson R
authors.initials: HR
authors.lastname: Granade
authors.forename: Kevin D
authors.initials: KD
authors.lastname: White
authors.forename: Robert W
authors.initials: RW
authors.lastname: Dickey
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000737:chemistry / Q000652:urine; D049872:Bivalvia / Q000378:metabolism*; D004141:Dinoflagellida; D004797:Enzyme-Linked Immunosorbent Assay; D005517:Foodborne Diseases; D008387:Marine Toxins / Q000737:chemistry / Q000506:poisoning* / Q000652:urine; D015394:Molecular Structure; D009498:Neurotoxins / Q000737:chemistry / Q000506:poisoning* / Q000652:urine; D035964:Oxocins / Q000737:chemistry / Q000506:poisoning* / Q000652:urine; D012758:Shellfish / Q000032:analysis; D057096:Shellfish Poisoning; D000095702:Polyether Toxins
chemical_list: D015415:Biomarkers; D008387:Marine Toxins; D009498:Neurotoxins; D035964:Oxocins; C053342:brevetoxin; D000095702:Polyether Toxins