{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T09:07:40Z","timestamp":1788858460310,"version":"build-2803163510"},"reference-count":64,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T00:00:00Z","timestamp":1788825600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003196","name":"Italian Ministry of Health","doi-asserted-by":"publisher","award":["RF-2021-12373885"],"award-info":[{"award-number":["RF-2021-12373885"]}],"id":[{"id":"10.13039\/501100003196","id-type":"DOI","asserted-by":"publisher"}]},{"award":["RF-2021-12373885"],"award-info":[{"award-number":["RF-2021-12373885"]}],"id":[{"id":"https:\/\/ror.org\/00789fa95","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Mar. Drugs","Marine Drugs"],"abstract":"<jats:p>Tetrodotoxin (TTX) is a potent marine neurotoxin responsible for severe seafood poisoning in humans, characterized by neurological symptoms that may be fatal. Originally identified as a natural contaminant of pufferfish (Tetraodontidae family), over the last few years TTX and its analogs have also been detected in other edible marine organisms, including mollusks, gastropods and crustaceans. Consequently, there is a need for rapid, sensitive, and reliable methods for TTX detection in seafood. In this study, a functional assay based on the use of mouse neuroblastoma Neuro-2a cells has been optimized and characterized for TTX detection in mussels. The assay is based on the toxin\u2019s ability to block voltage-gated sodium channels, thereby counteracting the sodium-dependent cytotoxicity induced by veratridine and ouabain. The linear range of the TTX standard curve fell between 0.44 and 33 ng\/mL, with limits of TTX detection (LOD) and quantitation (LOQ) of 0.132 ng\/mL and 0.439 ng\/mL, respectively, and good intra- and inter-day repeatability (RSDr= 15 and 11%, respectively). The assay also detected saxitoxin, which shares the same mechanism of action as TTX, but was less sensitive towards 4,9-anhydro-TTX. The minimum mussel extract dilution of 1:100 did not result in matrix-related interference, allowing accurate TTX quantitation, with a LOQ of 0.54 \u00b5g TTX equivalents\/kg mussel meat. Given its sensitivity, the optimized Neuro-2a assay represents a promising tool for toxicity-based TTX quantitation in mussels before their consumption.<\/jats:p>","DOI":"10.3390\/md24090314","type":"journal-article","created":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T08:10:22Z","timestamp":1788855022000},"page":"314","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Optimization and Intra-Laboratory Validation of the Neuro-2a Assay for Tetrodotoxin Detection in Mussels"],"prefix":"10.3390","volume":"24","author":[{"given":"Alessandra","family":"D\u2019Arelli","sequence":"first","affiliation":[{"name":"Department of Life Sciences, University of Trieste, Via A. Fleming 22, 34127 Trieste, Italy"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2909-6603","authenticated-orcid":false,"given":"Silvio","family":"Sosa","sequence":"additional","affiliation":[{"name":"Department of Life Sciences, University of Trieste, Via A. Fleming 22, 34127 Trieste, Italy"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"data-curation","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"conceptualization","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2120-6775","authenticated-orcid":false,"given":"Sonia","family":"Dall\u2019Ara","sequence":"additional","affiliation":[{"name":"National Reference Laboratory for Marine Biotoxins, Fondazione Centro Ricerche Marine, V.le A. Vespucci 2, 47042 Cesenatico, Italy"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"validation","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5361-4837","authenticated-orcid":false,"given":"Monica","family":"Cangini","sequence":"additional","affiliation":[{"name":"National Reference Laboratory for Marine Biotoxins, Fondazione Centro Ricerche Marine, V.le A. Vespucci 2, 47042 Cesenatico, Italy"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"validation","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3438-2196","authenticated-orcid":false,"given":"Michela","family":"Carlin","sequence":"additional","affiliation":[{"name":"Department of Life Sciences, University of Trieste, Via A. Fleming 22, 34127 Trieste, Italy"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9571-1993","authenticated-orcid":false,"given":"Pietro","family":"Antonelli","sequence":"additional","affiliation":[{"name":"Laboratory of Microbial Ecology and Genomics, Istituto Zooprofilattico Sperimentale delle Venezie, Viale dell\u2019Universit\u00e0 10, 35020 Legnaro, Italy"},{"name":"PhD National Programme in One Health Approaches to Infectious Diseases and Life Science Research, Department of Public Health, Experimental and Forensice Medicine, University of Pavia, 27100 Pavia, Italy"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"resources","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"}]},{"given":"Aurora","family":"Dall\u2019Occo","sequence":"additional","affiliation":[{"name":"Laboratory of Microbial Ecology and Genomics, Istituto Zooprofilattico Sperimentale delle Venezie, Viale dell\u2019Universit\u00e0 10, 35020 Legnaro, Italy"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"resources","vocabulary":"credit"}]},{"given":"Nicolas","family":"Scapin","sequence":"additional","affiliation":[{"name":"National Reference Center for fish, Mollusc and Crustacean Diseaseas, Istituto Zooprofilattico Sperimentale delle Venezie, Viale dell\u2019Universit\u00e0 10, 35020 Legnaro, Italy"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"resources","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7989-1317","authenticated-orcid":false,"given":"Giuseppe","family":"Arcangeli","sequence":"additional","affiliation":[{"name":"National Reference Center for fish, Mollusc and Crustacean Diseaseas, Istituto Zooprofilattico Sperimentale delle Venezie, Viale dell\u2019Universit\u00e0 10, 35020 Legnaro, Italy"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"resources","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8234-1292","authenticated-orcid":false,"given":"Carmen","family":"Losasso","sequence":"additional","affiliation":[{"name":"Laboratory of Microbial Ecology and Genomics, Istituto Zooprofilattico Sperimentale delle Venezie, Viale dell\u2019Universit\u00e0 10, 35020 Legnaro, Italy"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"resources","vocabulary":"credit"}]},{"given":"Aurelia","family":"Tubaro","sequence":"additional","affiliation":[{"name":"Department of Life Sciences, University of Trieste, Via A. Fleming 22, 34127 Trieste, Italy"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4306-7411","authenticated-orcid":false,"given":"Marco","family":"Pelin","sequence":"additional","affiliation":[{"name":"Department of Life Sciences, University of Trieste, Via A. Fleming 22, 34127 Trieste, Italy"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,8]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1081\/TXR-100103080","article-title":"Puffer Poisoning: Epidemiology and Treatment","volume":"20","author":"Noguchi","year":"2001","journal-title":"J. Toxicol. Toxin Rev."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"219","DOI":"10.1016\/S1474-4422(05)70041-7","article-title":"Neurotoxic Marine Poisoning","volume":"4","author":"Isbister","year":"2005","journal-title":"Lancet Neurol."},{"key":"ref_3","first-page":"e04752","article-title":"Risks for Public Health Related to the Presence of Tetrodotoxin (TTX) and TTX Analogues in Marine Bivalves and Gastropods","volume":"15","author":"Knutsen","year":"2017","journal-title":"EFSA J."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/S0006-3495(94)80746-5","article-title":"A Structural Model of the Tetrodotoxin and Saxitoxin Binding Site of the Na+ Channel","volume":"66","author":"Lipkind","year":"1994","journal-title":"Biophys. J."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1081\/TXR-100102537","article-title":"Pharmacology of Tetrodotoxin","volume":"20","author":"Narahashi","year":"2001","journal-title":"J. Toxicol. Toxin Rev."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1016\/j.toxicon.2012.11.026","article-title":"The Molecular Mystique of Tetrodotoxin","volume":"63","author":"Moczydlowski","year":"2013","journal-title":"Toxicon"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Magarlamov, T.Y., Melnikova, D.I., and Chernyshev, A.V. (2017). Tetrodotoxin-Producing Bacteria: Detection, Distribution and Migration of the Toxin in Aquatic Systems. Toxins, 9.","DOI":"10.3390\/toxins9050166"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Katikou, P., Gokbulut, C., Kosker, A.R., Camp\u00e0s, M., and Ozogul, F. (2022). An Updated Review of Tetrodotoxin and Its Peculiarities. Mar. Drugs, 20.","DOI":"10.3390\/md20010047"},{"key":"ref_9","first-page":"213","article-title":"Identification of Tetrodotoxin in Marine Gastropods Implicated in Food Poisoning","volume":"11","author":"Sui","year":"2002","journal-title":"J. Nat. Toxins"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/S0041-0101(02)00169-1","article-title":"Intra-Tissue Distribution of Tetrodotoxin in Two Marine Puffers Takifugu vermicularis and Chelonodon patoca","volume":"41","author":"Mahmud","year":"2003","journal-title":"Toxicon"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1153","DOI":"10.1016\/j.foodchem.2006.10.021","article-title":"Tetrodotoxin-Binding Proteins Isolated from Five Species of Toxic Gastropods","volume":"103","author":"Hwang","year":"2007","journal-title":"Food Chem."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1016\/j.aquatox.2011.04.001","article-title":"On the Origins and Biosynthesis of Tetrodotoxin","volume":"104","author":"Chau","year":"2011","journal-title":"Aquat. Toxicol."},{"key":"ref_13","first-page":"276939","article-title":"Tetrodotoxin Poisoning Due to Pufferfish and Gastropods, and Their Intoxication Mechanism","volume":"2011","author":"Noguchi","year":"2011","journal-title":"Int. Sch. Res. Not."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1307","DOI":"10.1016\/j.toxicon.2012.08.015","article-title":"Microdistribution of Tetrodotoxin in Two Species of Blue-Ringed Octopuses (Hapalochlaena lunulata and Hapalochlaena fasciata) Detected by Fluorescent Immunolabeling","volume":"60","author":"Williams","year":"2012","journal-title":"Toxicon"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"124404","DOI":"10.1016\/j.chemosphere.2019.124404","article-title":"Tetrodotoxin in Marine Bivalves and Edible Gastropods: A Mini-Review","volume":"236","author":"Biessy","year":"2019","journal-title":"Chemosphere"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"964","DOI":"10.1016\/j.toxicon.2008.10.001","article-title":"Lessepsian Migration and Tetrodotoxin Poisoning Due to Lagocephalus sceleratus in the Eastern Mediterranean","volume":"52","author":"Bentur","year":"2008","journal-title":"Toxicon"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"5622","DOI":"10.1021\/ac800769e","article-title":"First Toxicity Report of Tetrodotoxin and 5,6,11-TrideoxyTTX in the Trumpet Shell Charonia lampas lampas in Europe","volume":"80","author":"Rodriguez","year":"2008","journal-title":"Anal. Chem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"612","DOI":"10.1016\/j.jemermed.2008.09.024","article-title":"Seafood Intoxication by Tetrodotoxin: First Case in Europe","volume":"39","author":"Loureo","year":"2010","journal-title":"J. Emerg. Med."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1103","DOI":"10.1016\/j.foodchem.2011.11.081","article-title":"Liquid Chromatography\u2013Mass Spectrometry Method to Detect Tetrodotoxin and Its Analogues in the Puffer Fish Lagocephalus sceleratus (Gmelin, 1789) from European Waters","volume":"132","author":"Alfonso","year":"2012","journal-title":"Food Chem."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"21009","DOI":"10.2807\/1560-7917.ES2015.20.2.21009","article-title":"Detection of the Pufferfish Toxin Tetrodotoxin in European Bivalves, England, 2013 to 2014","volume":"20","author":"Turner","year":"2015","journal-title":"Eurosurveillance"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1779","DOI":"10.3390\/toxins7051779","article-title":"First Detection of Tetrodotoxin in Greek Shellfish by UPLC-MS\/MS Potentially Linked to the Presence of the Dinoflagellate Prorocentrum minimum","volume":"7","author":"Vlamis","year":"2015","journal-title":"Toxins"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Gerssen, A., Bovee, T.H.F., Klijnstra, M.D., Poelman, M., Portier, L., and Hoogenboom, R.L.A.P. (2018). First Report on the Occurrence of Tetrodotoxins in Bivalve Mollusks in The Netherlands. Toxins, 10.","DOI":"10.3390\/toxins10110450"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1016\/j.chemosphere.2018.12.010","article-title":"Tetrodotoxin Levels of Three Pufferfish Species (Lagocephalus sp.) Caught in the North-Eastern Mediterranean Sea","volume":"219","author":"Kosker","year":"2019","journal-title":"Chemosphere"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Hort, V., Arnich, N., Gu\u00e9rin, T., Lavison-Bompard, G., and Nicolas, M. (2020). First Detection of Tetrodotoxin in Bivalves and Gastropods from the French Mainland Coasts. Toxins, 12.","DOI":"10.3390\/toxins12090599"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Costa, P.R., Gir\u00e1ldez, J., Rodrigues, S.M., Le\u00e3o, J.M., Pinto, E., Soli\u00f1o, L., and Gago-Mart\u00ednez, A. (2021). High Levels of Tetrodotoxin (TTX) in Trumpet Shell Charonia lampas from the Portuguese Coast. Toxins, 13.","DOI":"10.3390\/toxins13040250"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"107510","DOI":"10.1016\/j.foodcont.2020.107510","article-title":"First Occurrence of Tetrodotoxins in Bivalve Mollusks from Northern Adriatic Sea (Italy)","volume":"120","author":"Bordin","year":"2021","journal-title":"Food Control"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"719","DOI":"10.1111\/1541-4337.12881","article-title":"Tetrodotoxin in Live Bivalve Mollusks from Europe: Is It to Be Considered an Emerging Concern for Food Safety?","volume":"21","author":"Antonelli","year":"2022","journal-title":"Compr. Rev. Food Sci. Food Saf."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Bacchiocchi, S., Campacci, D., Siracusa, M., Dubbini, A., Accoroni, S., Romagnoli, T., Campanelli, A., Griffoni, F., Tavoloni, T., and Gorbi, S. (2023). A Hotspot of TTX Contamination in the Adriatic Sea: Study on the Origin and Causative Factors. Mar. Drugs, 21.","DOI":"10.3390\/md21010008"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"50","DOI":"10.1016\/j.toxicon.2009.03.012","article-title":"First Report on Toxicity Assessment of the Lessepsian Migrant Pufferfish Lagocephalus sceleratus (Gmelin, 1789) from European Waters (Aegean Sea, Greece)","volume":"54","author":"Katikou","year":"2009","journal-title":"Toxicon"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"881","DOI":"10.1016\/j.chemosphere.2018.10.081","article-title":"First Detection of Tetrodotoxin and High Levels of Paralytic Shellfish Poisoning Toxins in Shellfish from Sicily (Italy) by Three Different Analytical Methods","volume":"215","author":"Tartaglione","year":"2019","journal-title":"Chemosphere"},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Bacchiocchi, S., Campacci, D., Siracusa, M., Dubbini, A., Leoni, F., Tavoloni, T., Accoroni, S., Gorbi, S., Giuliani, M.E., and Stramenga, A. (2021). Tetrodotoxins (TTXs) and Vibrio Alginolyticus in Mussels from Central Adriatic Sea (Italy): Are They Closely Related?. Mar. Drugs, 19.","DOI":"10.3390\/md19060304"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Bacchiocchi, S., Siracusa, M., Diomedi, G., Mazza, S., Calandri, E., Tavoloni, T., Vivani, V., Cangini, M., Arcangeli, G., and Losasso, C. (2025). Survey of Tetrodotoxins (TTXs) in Gastropods, Sea Urchins, and Blue Crabs from the Adriatic Sea: First Report in Paracentrotus lividus. Foods, 14.","DOI":"10.3390\/foods14234036"},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Botana, L.M., Alfonso, A., Rodr\u00edguez, I., Botana, A.M., Louzao, M.D.C., and Vieytes, M.R. (2016). How Safe Is Safe for Marine Toxins Monitoring?. Toxins, 8.","DOI":"10.3390\/toxins8070208"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Katikou, P. (2019). Public Health Risks Associated with Tetrodotoxin and Its Analogues in European Waters: Recent Advances after The EFSA Scientific Opinion. Toxins, 11.","DOI":"10.3390\/toxins11050240"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"579","DOI":"10.2478\/jvetres-2019-0060","article-title":"Marine Tetrodotoxin as a Risk for Human Health","volume":"63","author":"Madejska","year":"2019","journal-title":"J. Vet. Res."},{"key":"ref_36","unstructured":"(2025, October 01). Regulation\u2014854\/2004\u2014EN\u2014EUR-Lex. Available online: https:\/\/eur-lex.europa.eu\/eli\/reg\/2004\/854\/oj\/eng."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"693","DOI":"10.3390\/toxins6020693","article-title":"Tetrodotoxin: Chemistry, Toxicity, Source, Distribution and Detection","volume":"6","author":"Bane","year":"2014","journal-title":"Toxins"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"5719","DOI":"10.3390\/md12125719","article-title":"Alternative Methods for the Detection of Emerging Marine Toxins: Biosensors, Biochemical Assays and Cell-Based Assays","volume":"12","author":"Carnicer","year":"2014","journal-title":"Mar. Drugs"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"12249","DOI":"10.1021\/acs.jafc.2c04106","article-title":"Tetrodotoxin and Its Analogues in Food: Recent Updates on Sample Preparation and Analytical Methods Since 2012","volume":"70","author":"Hu","year":"2022","journal-title":"J. Agric. Food Chem."},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Lin, C., Li, Q., Liu, D., Feng, Q., Zhou, H., Shi, B., Zhang, X., Hu, Y., Jiang, X., and Sun, X. (2024). Recent Research Progress in Tetrodotoxin Detection and Quantitative Analysis Methods. Front. Chem., 12.","DOI":"10.3389\/fchem.2024.1447312"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1081\/TXR-100102535","article-title":"Current methodologies for detection of tetrodotoxin","volume":"20","author":"Noguchi","year":"2001","journal-title":"J. Toxicol. Toxin Rev."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Mi, W., and Liu, S. (2024). Tetrodotoxin and the State-of-the-Art Progress of Its Associated Analytical Methods. Front. Microbiol., 15.","DOI":"10.3389\/fmicb.2024.1413741"},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Zhang, X., Qiao, K., Cui, R., Xu, M., Cai, S., Huang, Q., and Liu, Z. (2024). Tetrodotoxin: The State-of-the-Art Progress in Characterization, Detection, Biosynthesis, and Transport Enrichment. Mar. Drugs, 22.","DOI":"10.3390\/md22120531"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"190","DOI":"10.1006\/abio.1993.1476","article-title":"Tetrazolium-Based Cell Bioassay for Neurotoxins Active on Voltage-Sensitive Sodium Channels: Semiautomated Assay for Saxitoxins, Brevetoxins, and Ciguatoxins","volume":"214","author":"Manger","year":"1993","journal-title":"Anal. Biochem."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"2647","DOI":"10.1016\/S0006-3495(98)77710-0","article-title":"Differences in Saxitoxin and Tetrodotoxin Binding Revealed by Mutagenesis of the Na+ Channel Outer Vestibule","volume":"75","author":"Penzotti","year":"1998","journal-title":"Biophys. J."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1016\/0041-0101(88)90171-7","article-title":"A Tissue Culture Assay for Tetrodotoxin, Saxitoxin and Related Toxins","volume":"26","author":"Kogure","year":"1988","journal-title":"Toxicon"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1016\/j.toxicon.2016.05.012","article-title":"The Voltage-Gated Sodium Ion Channel Inhibitory Activities of a New Tetrodotoxin Analogue, 4,4a-Anhydrotetrodotoxin, and Three Other Analogues Evaluated by Colorimetric Cell-Based Assay","volume":"119","author":"Saruhashi","year":"2016","journal-title":"Toxicon"},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Alkassar, M., Sanchez-Henao, A., Revert\u00e9, J., Barreiro, L., Rambla-Alegre, M., Leonardo, S., Mandalakis, M., Peristeraki, P., Diog\u00e8ne, J., and Camp\u00e0s, M. (2023). Evaluation of Toxicity Equivalency Factors of Tetrodotoxin Analogues with a Neuro-2a Cell-Based Assay and Application to Puffer Fish from Greece. Mar. Drugs, 21.","DOI":"10.3390\/md21080432"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"541","DOI":"10.1016\/j.toxicon.2008.06.028","article-title":"Comparative Study of the Use of Neuroblastoma Cells (Neuro-2a) and Neuroblastoma \u00d7 Glioma Hybrid Cells (NG108-15) for the Toxic Effect Quantification of Marine Toxins","volume":"52","year":"2008","journal-title":"Toxicon"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"381","DOI":"10.1016\/j.toxicon.2009.08.017","article-title":"Improvements in the Use of Neuroblastoma \u00d7 Glioma Hybrid Cells (NG108-15) for the Toxic Effect Quantification of Marine Toxins","volume":"55","year":"2010","journal-title":"Toxicon"},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Viallon, J., Chinain, M., and Darius, H.T. (2020). Revisiting the Neuroblastoma Cell-Based Assay (CBA-N2a) for the Improved Detection of Marine Toxins Active on Voltage Gated Sodium Channels (VGSCs). Toxins, 12.","DOI":"10.3390\/toxins12050281"},{"key":"ref_52","doi-asserted-by":"crossref","unstructured":"Ghasemi, M., Turnbull, T., Sebastian, S., and Kempson, I. (2021). The MTT Assay: Utility, Limitations, Pitfalls, and Interpretation in Bulk and Single-Cell Analysis. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms222312827"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"725","DOI":"10.1016\/0041-0101(85)90002-9","article-title":"Actions of 4-EpiTetrodotoxin and Anhydrotetrodotoxin on the Squid Axon","volume":"23","author":"Kao","year":"1985","journal-title":"Toxicon"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"C783","DOI":"10.1152\/ajpcell.00070.2007","article-title":"The TTX Metabolite 4,9-Anhydro-TTX Is a Highly Specific Blocker of the Na(v1.6) Voltage-Dependent Sodium Channel","volume":"293","author":"Rosker","year":"2007","journal-title":"Am. J. Physiol. Cell Physiol."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"865","DOI":"10.1021\/acs.chemrestox.0c00494","article-title":"Targeting Chloride Ion Channels: New Insights into the Mechanism of Action of the Marine Toxin Azaspiracid","volume":"34","author":"Louzao","year":"2021","journal-title":"Chem. Res. Toxicol."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"82","DOI":"10.3390\/md403082","article-title":"Ciguatoxins: Cyclic Polyether Modulators of Voltage-Gated Iion Channel Function","volume":"4","author":"Nicholson","year":"2006","journal-title":"Mar. Drugs"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"101994","DOI":"10.1016\/j.hal.2021.101994","article-title":"Improving In Vitro Ciguatoxin and Brevetoxin Detection: Selecting Neuroblastoma (Neuro-2a) Cells with Lower Sensitivity to Ouabain and Veratridine (OV-LS)","volume":"103","author":"Loeffler","year":"2021","journal-title":"Harmful Algae"},{"key":"ref_58","doi-asserted-by":"crossref","unstructured":"Litaker, R.W., Holland, W.C., Hardison, D.R., Pisapia, F., Hess, P., Kibler, S.R., and Tester, P.A. (2017). Ciguatoxicity of Gambierdiscus and Fukuyoa Species from the Caribbean and Gulf of Mexico. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0185776"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"576","DOI":"10.1016\/j.scitotenv.2019.03.445","article-title":"Predictive Score and Probability of CTX-like Toxicity in Fish Samples from the Official Control of Ciguatera in the Canary Islands","volume":"673","author":"Saavedra","year":"2019","journal-title":"Sci. Total Environ."},{"key":"ref_60","unstructured":"(2025, September 23). Eurachem Guides. Available online: https:\/\/www.eurachem.org\/index.php\/publications\/guides\/."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"659","DOI":"10.1016\/j.talanta.2017.08.058","article-title":"Development and Validation of a Maleimide-Based Enzyme-Linked Immunosorbent Assay for the Detection of Tetrodotoxin in Oysters and Mussels","volume":"176","author":"Leonardo","year":"2018","journal-title":"Talanta"},{"key":"ref_62","unstructured":"(2026, August 31). AESAN\u2014Laboratorio de Biotoxinas Marinas\/EU Reference Laboratory for Marine Biotoxins (EURLMB) Determination of Tetrodotoxin by HILIC-MS\/MS. Available online: https:\/\/www.aesan.gob.es\/en\/laboratorios\/eurlmb\/documentos-publicos."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"139682","DOI":"10.1016\/j.chemosphere.2023.139682","article-title":"Tetrodotoxin in Bivalve Mollusks: An Integrated Study towards the Comprehension of the Influencing Factors of a Newly Native Phenomenon","volume":"339","author":"Antonelli","year":"2023","journal-title":"Chemosphere"},{"key":"ref_64","unstructured":"(2020). Ultrahigh-Performance Hydrophilic Interaction Liquid Chromatography with Tandem Mass Spectrometry Method for the Determination of Paralytic Shellfish Toxins and Tetrodotoxin in Mussels, Oysters, Clams, Cockles, and Scallops: Collaborative Study. J. AOAC Int., 103, 533\u2013562."}],"container-title":["Marine Drugs"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1660-3397\/24\/9\/314\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T08:14:01Z","timestamp":1788855241000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1660-3397\/24\/9\/314"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,8]]},"references-count":64,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2026,9]]}},"alternative-id":["md24090314"],"URL":"https:\/\/doi.org\/10.3390\/md24090314","relation":{},"ISSN":["1660-3397"],"issn-type":[{"value":"1660-3397","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,8]]}}}