Cell-based assay coupled with chromatographic fractioning: A strategy for marine toxins detection in natural samples

Cell-based assays (CBA) have been proposed for the evaluation of toxicity caused by marine toxins in natural samples (fish, shellfish and microalgae). However, their application has been hindered due to the interferences present in biological matrices that may cause cellular response and interfere in toxicity evaluation. This work reviews in an extensive introduction the use of CBA for toxicity evaluation of marine toxins. Afterwards, the coupling of chromatographic fractioning with neuroblastoma Neuro-2a CBA is presented to enhance the applicability of CBA for complex matrices. Examples of application are provided for mussel samples (Mytilus galloprovincialis) and microalgae (Gambierdiscus sp.), and the results demonstrated the great potential of the combined strategy for reliable toxicological evaluation without ethical concern. Fractioning of an equivalent of 72 mg eq mL^?1 of mussel sample allowed the identification of non-toxic and toxic fractions whereas only 2.5 mg eq mL^?1 of non-purified mussel sample was responsible for 20% of cell mortality. Furthermore, the application of CBA allowed selectively distinguishing between ciguatoxin-like and other unspecific toxicity in Gambierdiscus sp. extract.