title: Multi-Laboratory Study of Five Methods for the Determination of Brevetoxins in Shellfish Tissue Extracts.
abstract: A thirteen-laboratory comparative study tested the performance of four methods as alternatives to mouse bioassay for the determination of brevetoxins in shellfish. The methods were N2a neuroblastoma cell assay, two variations of the sodium channel receptor binding assay, competitive ELISA, and LC/MS. Three to five laboratories independently performed each method using centrally prepared spiked and naturally incurred test samples. Competitive ELISA and receptor binding (96-well format) compared most favorably with mouse bioassay. Between-laboratory relative standard deviations (RSDR) ranged from 10 to 20% for ELISA and 14 to 31% for receptor binding. Within-laboratory (RSDr) ranged from 6 to 15% for ELISA, and 5 to 31% for receptor binding. Cell assay was extremely sensitive but data variation rendered it unsuitable for statistical treatment. LC/MS performed as well as ELISA on spiked test samples but was inordinately affected by lack of toxin-metabolite standards, uniform instrumental parameters, or both, on incurred test samples. The ELISA and receptor binding assay are good alternatives to mouse bioassay for the determination of brevetoxins in shellfish.
authors.affiliation: CFSAN, GCSL, USFDA, 1 Iberville Drive, Dauphin Island, AL 36528, USA.
authors.forename: Robert W
authors.initials: RW
authors.lastname: Dickey
authors.affiliation: CFSAN, USFDA, 8301 Muirkirk Road, Laurel, MD 20708, USA.
authors.forename: Steven M
authors.initials: SM
authors.lastname: Plakas
authors.affiliation: Florida FWCC, Florida Marine Research Institute, 100 8th Ave.SE, St. Petersburg, FL 33701, USA.
authors.forename: Edward L E
authors.initials: EL
authors.lastname: Jester
authors.affiliation: NOAA, National Ocean Service, 219 Fort Johnson Road, Charleston, SC 29412, USA.
authors.forename: Kathleen R
authors.initials: KR
authors.lastname: El Said
authors.affiliation: Mote Marine Laboratory, 1600 Ken Thompson Parkway, Sarasota, FL 34236, USA.
authors.forename: Jan N
authors.initials: JN
authors.lastname: Johannessen
authors.affiliation: USAMRIID, Toxinology Division, 1425 Porter Street, Fort Detrick, MD 21702, USA.
authors.forename: Leanne J
authors.initials: LJ
authors.lastname: Flewelling
authors.affiliation: USEPA, Gulf Ecology Division, 1 Sabine Island Drive, Gulf Breeze, FL 32561, USA.
authors.forename: Paula
authors.initials: P
authors.lastname: Scott
authors.affiliation: UNCW, Center for Marine Science, 5600 Marvin K. Moss Lane, Wilmington, NC 28409, USA.
authors.forename: Dan G
authors.initials: DG
authors.lastname: Hammond
authors.affiliation: CFSAN, OSAS, USFDA, 5100 Paint Branch Parkway, College Park, MD 20740, USA.
authors.forename: Frances M
authors.initials: FM
authors.lastname: Van Dolah
authors.affiliation: ESR, Kenepuru Drive, Porirua, New Zealand.
authors.forename: Tod A
authors.initials: TA
authors.lastname: Leighfield
authors.affiliation: Crop and Food Research, Christchurch, New Zealand.
authors.forename: Marie-Yasmine
authors.initials: MY
authors.lastname: Bottein Dachraoui
authors.affiliation: ORA, Northeast Regional Laboratory, USFDA, 158-15 Liberty Ave., Jamaica, NY 11433, USA.
authors.forename: John S
authors.initials: JS
authors.lastname: Ramsdell
authors.affiliation: NRC Institute of Marine Biosciences, 1411 Oxford Street, Halifax, NS Canada B3H3Z1.
authors.forename: Richard H
authors.initials: RH
authors.lastname: Pierce
authors.affiliation: CFSAN, OSAS Division of Mathematics, USFDA, 1425 Paint Branch Parkway, College Park, MD 20740, USA.
authors.forename: Mike S
authors.initials: MS
authors.lastname: Henry
authors.forename: Mark A
authors.initials: MA
authors.lastname: Poli
authors.forename: Calvin
authors.initials: C
authors.lastname: Walker
authors.forename: Jan
authors.initials: J
authors.lastname: Kurtz
authors.forename: Jerome
authors.initials: J
authors.lastname: Naar
authors.forename: Daniel G
authors.initials: DG
authors.lastname: Baden
authors.forename: Steve M
authors.initials: SM
authors.lastname: Musser
authors.forename: Kevin D
authors.initials: KD
authors.lastname: White
authors.forename: Penelope
authors.initials: P
authors.lastname: Truman
authors.forename: Aaron
authors.initials: A
authors.lastname: Miller
authors.forename: Timothy P
authors.initials: TP
authors.lastname: Hawryluk
authors.forename: Marleen M
authors.initials: MM
authors.lastname: Wekell
authors.forename: David
authors.initials: D
authors.lastname: Stirling
authors.forename: Michael A
authors.initials: MA
authors.lastname: Quilliam
authors.forename: Jung K
authors.initials: JK
authors.lastname: Lee