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.
Nozawa
Akira
A
Pharmaceutical R and D Department, POLA Chemical Industries, Inc., 560 Kashio-cho, Totsuka-ku, Yokohama 244-0812, Japan.
Tsuji
Kuniro
K
Ishida
Hitoshi
H
D000818:Animals; D001681:Biological Assay; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004141:Dinoflagellida; D004305:Dose-Response Relationship, Drug; D005517:Foodborne Diseases / Q000209:etiology; D008387:Marine Toxins / Q000302:isolation & purification* / Q000633:toxicity; D051379:Mice; D008974:Mollusca; D009498:Neurotoxins / Q000302:isolation & purification / Q000633:toxicity; D009520:New Zealand; D035964:Oxocins / Q000302:isolation & purification* / Q000633:toxicity; D012604:Scorpion Venoms / Q000302:isolation & purification* / Q000633:toxicity; D012758:Shellfish / Q000032:analysis* / Q000633:toxicity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D000095702:Polyether Toxins
D008387:Marine Toxins; D009498:Neurotoxins; D035964:Oxocins; D012604:Scorpion Venoms; C454815:parabutoxin 3 protein, Parabuthus transvaalicus; C043466:brevetoxin B; D000095702:Polyether Toxins