Cyclodextrin polymer clean-up method for the detection of ciguatoxins in fish with cell-based assays

Identificador:  imarina:9282207

Handle: https://hdl.handle.net/20.500.11797/imarina9282207

Autors:  Campas, Monica; Leonardo, Sandra; Rambla-Alegre, Maria; Sagrista, Nuria; Vaya, Raquel; Diogene, Jorge; Torrens, Mabel; Fragoso, Alex

Resum:
Ciguatoxins (CTXs) are marine toxins produced by microalgae of the genera Gambierdiscus and Fukuyoa, which are transferred through the food webs, reaching humans and causing a poisoning known as ciguatera. The cell-based assay (CBA) is commonly used for their detection because of its high sensitivity and the provided toxicological information. However, matrix effects may interfere in the CBA. In this work, γ-cyclodextrin-hexamethylene diisocyanate (γ-CD-HDI), γ-cyclodextrin-epichlorohydrin (γ-CD-EPI) and γ-CD-EPI conjugated to magnetic beads (γ-CD-EPI-MB) have been evaluated as clean-up materials for fish flesh extracts containing CTXs. The best results were achieved with γ-CD-HDI in column format, which showed a CTX1B recovery of 42% and 32% for Variola louti and Seriola dumerili, respectively, and allowed exposing cells to at least 400 mg/mL of fish flesh. This clean-up strategy provides at least 4.6 and 3.0-fold higher sensitivities to the assay for V. louti and S. dumerili, respectively, improving the reliability of CTX quantification.