Implication of brevetoxin B1 and PbTx-3 in neurotoxic shellfish poisoning in New Zealand by isolation and quantitative determination with liquid chromatography-tandem mass spectrometry.

Brevetoxin B1 (BTX-B1) was isolated from Austrovenus stutchburyi following the 1992-1993 outbreak of neurotoxic shellfish poisoning (NSP) in New Zealand. We report here the first isolation of PbTx-3 from the same shellfish and the development of a procedure for quantitative determination of PbTx-3 and BTX-B1. PbTx-3 was isolated by chromatography on columns of SiO2, ODS, and LH-20, followed by reverse-phase HPLCs. In mass spectrometry (MS) with an electrospray ionization (ESI) interface operating in the positive or negative ion mode, the abundant protonated ion [M+H]+ of PbTx-3 (m/z 897) and the de-sodiated ion [M-Na]- of BTX-B1 (m/z 1016) were generated, respectively. These served as precursor ions for collision-induced dissociation, and the product ions of m/z 725 from PbTx-3 and m/z 80 from BTX-B1 were identified, allowing unambiguous confirmation of these toxins by selected reaction monitoring liquid chromatography-tandem mass spectrometry (SRM LC-MS/MS) analysis. The determination limits were 0.4 and 2 ng/g for BTX-B1 and PbTx-3 at a signal-to-noise ratio of five, respectively. This LC-MS/MS method was successfully applied to determine BTX-B1 and PbTx-3 in the NSP-associated toxic shellfish. BTX-B1 was found in both A. stutchburyi and Perna canaliculus, but not in Crassostrea gigas, while PbTx-3 was found in all three.
AuthorsAkira Nozawa, Kuniro Tsuji, Hitoshi Ishida
JournalToxicon : official journal of the International Society on Toxinology (Toxicon) Vol. 42 Issue 1 Pg. 91-103 (Jul 2003) ISSN: 0041-0101 [Print] England
PMID12893066 (Publication Type: Journal Article)
Chemical References
  • Marine Toxins
  • Neurotoxins
  • Oxocins
  • Scorpion Venoms
  • parabutoxin 3 protein, Parabuthus transvaalicus
  • brevetoxin B
  • Animals
  • Biological Assay
  • Chromatography, High Pressure Liquid (methods)
  • Dinoflagellida
  • Dose-Response Relationship, Drug
  • Foodborne Diseases (etiology)
  • Marine Toxins (isolation & purification, toxicity)
  • Mice
  • Mollusca
  • Neurotoxins (isolation & purification, toxicity)
  • New Zealand
  • Oxocins (isolation & purification, toxicity)
  • Scorpion Venoms (isolation & purification, toxicity)
  • Shellfish (analysis, toxicity)
  • Spectrometry, Mass, Electrospray Ionization (methods)

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