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This research was funded by the Ministerio de Ciencia e Innovación (MICIN) and the Agencia Estatal de Investigación (AEI) through the CELLECTRA (PID2020-112976RB-C21) project. Mounira Alkassar acknowledges MICIN and AEI for her PhD grant (PRE2019-088181). Jaume Reverté acknowledges IRTA for his PhD grant (CPI0422). Lourdes Barreiro acknowledges IRTA and URV for her PhD grant (2020PMF-PIPF-07). Andres Sanchez-Henao acknowledges the Canary Islands Research Agency (ACIISI) for his research staff training aid through the Catalina Ruiz Program (APCR2022010011). The authors also acknowledge the support from the CERCA Program/Generalitat de Catalunya.

Analysis of institutional authors

Alkassar, MAuthorReverte, JAuthorRambla-Alegre, MAuthorLeonardo, SAuthorDiogene, JAuthorCampas, MCorresponding Author

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August 11, 2023
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Article

Evaluation of Toxicity Equivalency Factors of Tetrodotoxin Analogues with a Neuro-2a Cell-Based Assay and Application to Puffer Fish from Greece

Publicated to:Marine Drugs. 21 (8): 432- - 2023-08-01 21(8), DOI: 10.3390/md21080432

Authors: Alkassar, Mounira; Sanchez-Henao, Andres; Reverte, Jaume; Barreiro, Lourdes; Rambla-Alegre, Maria; Leonardo, Sandra; Mandalakis, Manolis; Peristeraki, Panagiota; Diogene, Jorge; Campas, Monica

Affiliations

Aigües Marines i Continentals. IRTA Investigación y Tecnología Agroalimentarias - Author
Aigües Marines i Continentals. Producció Animal - Author
Hellen Ctr Marine Res, Inst Marine Biol Biotechnol & Aquaculture, Iraklion 71003, Greece - Author
Hellen Ctr Marine Res, Inst Marine Biol Resources & Inland Waters, Iraklion 71003, Greece - Author
Inst Recerca & Tecnol Agroalimentaries IRTA, Ctra Poble Nou Km 5-5, La Rapita 43540, Spain - Author
Producció Animal. IRTA Investigación y Tecnología Agroalimentarias - Author
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Abstract

Tetrodotoxin (TTX) is a potent marine neurotoxin involved in poisoning cases, especially through the consumption of puffer fish. Knowledge of the toxicity equivalency factors (TEFs) of TTX analogues is crucial in monitoring programs to estimate the toxicity of samples analyzed with instrumental analysis methods. In this work, TTX analogues were isolated from the liver of a Lagocephalus sceleratus individual caught on South Crete coasts. A cell-based assay (CBA) for TTXs was optimized and applied to the establishment of the TEFs of 5,11-dideoxyTTX, 11-norTTX-6(S)-ol, 11-deoxyTTX and 5,6,11-trideoxyTTX. Results showed that all TTX analogues were less toxic than the parent TTX, their TEFs being in the range of 0.75–0.011. Then, different tissues of three Lagocephalus sceleratus individuals were analyzed with CBA and liquid chromatography-tandem mass spectrometry (LC-MS/MS). The obtained TEFs were applied to the TTX analogues’ concentrations obtained by LC-MS/MS analysis, providing an indication of the overall toxicity of the sample. Information about the TEFs of TTX analogues is valuable for food safety control, allowing the estimation of the risk of fish products to consumers.

Keywords

1st detectioncaughtcell-based assay (cba)lagocephalus sceleratusliquid chromatography-tandem mass spectrometry (lc-ms/ms)mass-spectrometry methodneuroblastoma-cellspuffer fishsaxitoxinsodium-channeltetrodotoxin (ttx)tissue-culture bioassaytoxicity equivalency factor (tef)toxinsttxttx analogueAnimalsCell-based assay (cba)Chromatography, liquidGreeceLagocephalus sceleratusLagocephalus-sceleratus gmelinLiquid chromatography-tandem mass spectrometry (lc-ms/ms)Puffer fishTandem mass spectrometryTetraodontiformesTetrodotoxinTetrodotoxin (ttx)Toxicity equivalency factor (tef)Ttx analogue

Quality index

Bibliometric impact. Analysis of the contribution and dissemination channel

The work has been published in the journal Marine Drugs due to its progression and the good impact it has achieved in recent years, according to the agency WoS (JCR), it has become a reference in its field. In the year of publication of the work, 2023, it was in position 46/354, thus managing to position itself as a Q1 (Primer Cuartil), in the category Pharmacology & Pharmacy.

From a relative perspective, and based on the normalized impact indicator calculated from World Citations provided by WoS (ESI, Clarivate), it yields a value for the citation normalization relative to the expected citation rate of: 1.92. This indicates that, compared to works in the same discipline and in the same year of publication, it ranks as a work cited above average. (source consulted: ESI Nov 14, 2024)

This information is reinforced by other indicators of the same type, which, although dynamic over time and dependent on the set of average global citations at the time of their calculation, consistently position the work at some point among the top 50% most cited in its field:

  • Field Citation Ratio (FCR) from Dimensions: 5.06 (source consulted: Dimensions Sep 2025)

Specifically, and according to different indexing agencies, this work has accumulated citations as of 2025-09-09, the following number of citations:

  • WoS: 11
  • Europe PMC: 7

Impact and social visibility

From the perspective of influence or social adoption, and based on metrics associated with mentions and interactions provided by agencies specializing in calculating the so-called "Alternative or Social Metrics," we can highlight as of 2025-09-09:

  • The use, from an academic perspective evidenced by the Altmetric agency indicator referring to aggregations made by the personal bibliographic manager Mendeley, gives us a total of: 19.
  • The use of this contribution in bookmarks, code forks, additions to favorite lists for recurrent reading, as well as general views, indicates that someone is using the publication as a basis for their current work. This may be a notable indicator of future more formal and academic citations. This claim is supported by the result of the "Capture" indicator, which yields a total of: 19 (PlumX).

With a more dissemination-oriented intent and targeting more general audiences, we can observe other more global scores such as:

  • The Total Score from Altmetric: 0.5.
  • The number of mentions on the social network X (formerly Twitter): 1 (Altmetric).

It is essential to present evidence supporting full alignment with institutional principles and guidelines on Open Science and the Conservation and Dissemination of Intellectual Heritage. A clear example of this is:

  • The work has been submitted to a journal whose editorial policy allows open Open Access publication.
  • Assignment of a Handle/URN as an identifier within the deposit in the Institutional Repository: http://hdl.handle.net/20.500.12327/2351

Leadership analysis of institutional authors

This work has been carried out with international collaboration, specifically with researchers from: Greece.

There is a significant leadership presence as some of the institution’s authors appear as the first or last signer, detailed as follows: First Author (ALKASSAR, MOUNIRA) and Last Author (CAMPAS HOMS, MONICA).

the author responsible for correspondence tasks has been CAMPAS HOMS, MONICA.