Detection of gymnodimine-A and 13-desmethyl C spirolide phycotoxins by fluorescence polarization. - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue Analytical Chemistry Année : 2009

Detection of gymnodimine-A and 13-desmethyl C spirolide phycotoxins by fluorescence polarization.

Résumé

The gymnodimines and spirolides are phycotoxins classified into a heterogeneous group of marine biocompounds called cyclic imines. Although there is no clear evidence of their toxicity to humans, gymnodimines and spirolides are highly toxic to rodents and constitute a source of false positives in lipophilic toxin detection by the mouse bioassay. Using nicotinic acetylcholine receptor-enriched membranes of Torpedo, and fluorescent alpha-bungarotoxin, we developed a fluorescence polarization assay to detect and quantify gymnodimine-A and 13-desmethyl C spirolide. The presence of these cyclic imines in solution inhibited the interaction of fluorescent-labeled alpha-bungarotoxin with nicotinic acetylcholine receptors in a concentration-dependent manner. The sensitivity of the assay is in the order of nanomolar concentrations of gymnodimine and 13-desmethyl C spirolide. Okadaic acid, yessotoxin, and brevetoxin-2, three lipophilic marine toxins, did not interfere with this assay. A suitable extraction method in shellfish was also developed. The gymnodimine-A and 13-desmethyl C spirolide recovery rates of mussel matrix extraction with acetone/chloroform were 63.6% +/- 3.5% and 87.4% +/- 5.3%, respectively. In summary, this inhibition assay is capable of gymnodimine-A and 13-desmethyl C spirolide detection in mussel extracts with enough sensitivity and specificity to quantify these toxins in the range of 50-2000 microg/kg and 70-700 microg/kg of shellfish meat, respectively.
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Dates et versions

hal-00378092 , version 1 (23-04-2009)

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Natalia Vilariño, Eva S Fonfría, Jordi Molgó, Rómulo Aráoz, Luis M Botana. Detection of gymnodimine-A and 13-desmethyl C spirolide phycotoxins by fluorescence polarization.. Analytical Chemistry, 2009, 81 (7), pp.2708-14. ⟨10.1021/ac900144r⟩. ⟨hal-00378092⟩

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