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Cell-based assay coupled with chromatographic fractioning: a strategy for marine toxins detection in natural samples
A Caillaud1, E Cañete, P de la Iglesia
1IRTA, C. Poble Nou, Km 5.5, 43540 Sant Carles de la Ràpita, Spain.
Abstract:
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 2.5mg 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.
Insights
Cell-based assays (CBA) show promise for marine toxin evaluation. Combining chromatography with CBA enhances accuracy in complex samples like shellfish and microalgae, enabling reliable toxicological assessment.
Area of Science:
- Environmental Toxicology
- Marine Biology
- Analytical Chemistry
Background:
- Cell-based assays (CBA) are proposed for marine toxin evaluation in natural samples.
- Interferences in biological matrices hinder CBA accuracy for marine toxins.
- Existing methods face challenges in complex sample matrices.
Purpose of the Study:
- To review the application of CBA for marine toxin toxicity evaluation.
- To enhance CBA applicability for complex matrices by coupling with chromatographic fractioning.
- To demonstrate a reliable toxicological evaluation strategy for marine toxins.
Main Methods:
- Extensive literature review on CBA for marine toxin evaluation.
- Coupling chromatographic fractioning with neuroblastoma Neuro-2a CBA.
- Application to mussel (Mytilus galloprovincialis) and microalgae (Gambierdiscus sp.) samples.
Main Results:
- Chromatographic fractioning improved CBA applicability for complex matrices.
- Fractioning identified non-toxic and toxic fractions in mussel samples.
- CBA distinguished ciguatoxin-like from unspecific toxicity in microalgae extracts.
Conclusions:
- The combined strategy of chromatographic fractioning and CBA offers reliable toxicological evaluation.
- This approach minimizes ethical concerns associated with traditional toxicity testing.
- The method shows great potential for analyzing marine toxins in complex natural samples.
