{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,8]],"date-time":"2026-03-08T00:09:05Z","timestamp":1772928545654,"version":"3.50.1"},"reference-count":42,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2021,3,31]],"date-time":"2021-03-31T00:00:00Z","timestamp":1617148800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["(UID\/Multi\/04326\/2020)"],"award-info":[{"award-number":["(UID\/Multi\/04326\/2020)"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>Tetrodotoxin (TTX) is a potent neurotoxin, considered an emerging toxin in Europe where recently a safety limit of 44 \u00b5g TTX kg\u22121 was recommended by authorities. In this study, three specimens of the large gastropod trumpet shell Charonia lampas bought in a market in south Portugal were analyzed using a neuroblastoma cell (N2a) based assay and by LC-MS\/MS. N2a toxicity was observed in the viscera of two individuals analyzed and LC-MS\/MS showed very high concentrations of TTX (42.1 mg kg\u22121) and 4,9-anhydroTTX (56.3 mg kg\u22121). A third compound with m\/z 318 and structurally related with TTX was observed. In the edible portion, i.e., the muscle, toxin levels were below the EFSA recommended limit. This study shows that trumpet shell marine snails are seafood species that may reach the markets containing low TTX levels in the edible portion but containing very high levels of TTX in non-edible portion raising concerns regarding food safety if a proper evisceration is not carried out by consumers. These results highlight the need for better understanding TTX variability in this gastropod species, which is critical to developing a proper legal framework for resources management ensuring seafood safety, and the introduction of these gastropods in the markets.<\/jats:p>","DOI":"10.3390\/toxins13040250","type":"journal-article","created":{"date-parts":[[2021,3,31]],"date-time":"2021-03-31T10:24:33Z","timestamp":1617186273000},"page":"250","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":26,"title":["High Levels of Tetrodotoxin (TTX) in Trumpet Shell Charonia lampas from the Portuguese Coast"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6083-470X","authenticated-orcid":false,"given":"Pedro Reis","family":"Costa","sequence":"first","affiliation":[{"name":"IPMA\u2014Portuguese Institute of the Sea and Atmosphere, Av. Bras\u00edlia, 1449-006 Lisbon, Portugal"},{"name":"CCMAR\u2014Center of Marine Sciences, Campus of Gambelas, University of Algarve, 8005-139 Faro, Portugal"}]},{"given":"Jorge","family":"Gir\u00e1ldez","sequence":"additional","affiliation":[{"name":"Biomedical Research Center (CINBIO), Department of Analytical and Food Chemistry, Campus Universitario de Vigo, University of Vigo, 36310 Vigo, Spain"}]},{"given":"Susana Margarida","family":"Rodrigues","sequence":"additional","affiliation":[{"name":"IPMA\u2014Portuguese Institute of the Sea and Atmosphere, Av. Bras\u00edlia, 1449-006 Lisbon, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2260-5281","authenticated-orcid":false,"given":"Jos\u00e9 Manuel","family":"Le\u00e3o","sequence":"additional","affiliation":[{"name":"Biomedical Research Center (CINBIO), Department of Analytical and Food Chemistry, Campus Universitario de Vigo, University of Vigo, 36310 Vigo, Spain"}]},{"given":"Estefan\u00eda","family":"Pinto","sequence":"additional","affiliation":[{"name":"IPMA\u2014Portuguese Institute of the Sea and Atmosphere, Av. Bras\u00edlia, 1449-006 Lisbon, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8356-5688","authenticated-orcid":false,"given":"Luc\u00eda","family":"Soli\u00f1o","sequence":"additional","affiliation":[{"name":"IPMA\u2014Portuguese Institute of the Sea and Atmosphere, Av. Bras\u00edlia, 1449-006 Lisbon, Portugal"},{"name":"CCMAR\u2014Center of Marine Sciences, Campus of Gambelas, University of Algarve, 8005-139 Faro, Portugal"}]},{"given":"Ana","family":"Gago-Mart\u00ednez","sequence":"additional","affiliation":[{"name":"Biomedical Research Center (CINBIO), Department of Analytical and Food Chemistry, Campus Universitario de Vigo, University of Vigo, 36310 Vigo, Spain"}]}],"member":"1968","published-online":{"date-parts":[[2021,3,31]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"276939","DOI":"10.5402\/2011\/276939","article-title":"Tetrodotoxin poisoning due to pufferfish and gastropods, and their intoxication mechanism","volume":"2011","author":"Noguchi","year":"2011","journal-title":"ISRN Toxicol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"935","DOI":"10.2331\/suisan.47.935","article-title":"Occurrence of tetrodotoxin in a trumpet shell, \u201cBoshubora\u201d Charonia sauliae","volume":"47","author":"Narita","year":"1981","journal-title":"Bull. Jpn. Soc. Sci. Fish."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1173","DOI":"10.2331\/suisan.48.1173","article-title":"Tetrodotoxin in the starfish Astropecten polyacanthus; in association with toxification of a trumpet shell, \u201cboshubora\u201d Charonia sauliae","volume":"48","author":"Noguchi","year":"1982","journal-title":"Bull. Jap. Soc. Sci. Fish."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"645","DOI":"10.1080\/87559129.2019.1669162","article-title":"A Global Retrospective Study on Human Cases of Tetrodotoxin (TTX) Poisoning after Seafood Consumption","volume":"36","author":"Guardone","year":"2020","journal-title":"Food Rev. Int."},{"key":"ref_5","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_6","doi-asserted-by":"crossref","first-page":"965","DOI":"10.1085\/jgp.47.5.965","article-title":"Tetrodotoxin blockage of sodium conductance increase in excitation","volume":"47","author":"Narahashi","year":"1964","journal-title":"J. Gen. Physiol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1438","DOI":"10.1126\/science.157.3795.1438","article-title":"Saxitoxin and tetrodotoxin: Comparison of nerve blocking mechanism","volume":"157","author":"Narahashi","year":"1967","journal-title":"Science"},{"key":"ref_8","first-page":"997","article-title":"Tetrodotoxin, saxitoxin and their significance in the study of excitation phenomenon","volume":"18","author":"Kao","year":"1966","journal-title":"Pharm. Rev."},{"key":"#cr-split#-ref_9.1","unstructured":"European Commission (2004). Commission Regulation"},{"key":"#cr-split#-ref_9.2","unstructured":"(EC) No 853\/2004 of the European Parliament and of the Council of 29 April 2004 laying down specific hygiene rules for on the hygiene of foodstuffs. Off. J. Eur. Union, 139, 55-205."},{"key":"#cr-split#-ref_10.1","unstructured":"European Commission (2004). Commission Regulation"},{"key":"#cr-split#-ref_10.2","unstructured":"(EC) No 854\/2004 of the European Parliament and of the Council of 29 April 2004 laying down specific rules for the organisation of official controls on products of animal origin intended for human consumption. Off. J. Eur. Union, 226, 83-127."},{"key":"ref_11","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":"Alfonso","year":"2010","journal-title":"J. Emerg. Med."},{"key":"ref_12","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":"Alfonso","year":"2008","journal-title":"Anal. Chem."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"712","DOI":"10.3390\/md10040712","article-title":"New Gastropod Vectors and Tetrodotoxin Potential Expansion in Temperate Waters of the Atlantic Ocean","volume":"10","author":"Silva","year":"2012","journal-title":"Mar. Drugs"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2","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_15","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_16","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 molluscs in the Netherlands. Toxins, 10.","DOI":"10.3390\/toxins10110450"},{"key":"ref_17","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_18","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_19","doi-asserted-by":"crossref","unstructured":"EFSA (2017). Scientific opinion on the risks for public health related to the presence of tetrodotoxin (TTX) and TTX analogues in marine bivalves and gastropods. EFSA J., 15, 4752.","DOI":"10.2903\/j.efsa.2017.4752"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Leao, J.M., Lozano-Leon, A., Gir\u00e1ldez, J., Vilari\u00f1o, O., and Gago-Mart\u00ednez, A. (2018). Preliminary results on the evaluation of the occurrence of tetrodotoxin associated to marine Vibrio spp in bivalves from the Galician Rias (Northwest Spain). Mar. Drugs, 16.","DOI":"10.3390\/md16030081"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Blanco, L., Lago, J., Gonz\u00e1lez, V., Paz, B., Rambla-Alegre, M., and Cabado, A.G. (2019). Occurrence of tetrodotoxin in bivalves and gastropods from harvesting areas and other natural spaces in Spain. Toxins, 11.","DOI":"10.3390\/toxins11060331"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Rodrigues, S.M., Pinto, E.P., Oliveira, P., Pedro, S., and Costa, P.R. (2019). Evaluation of the occurrence of tetrodotoxin in bivalve mollusks from the Portuguese coast. J. Mar. Sci. Eng., 7.","DOI":"10.3390\/jmse7070232"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"104780","DOI":"10.1016\/j.marenvres.2019.104780","article-title":"Tetrodotoxin and saxitoxin in two native species of puffer fish, Sphoeroides marmoratus and Lagocephalus lagocephalus, from NE Atlantic Ocean (Madeira Island, Portugal)","volume":"151","author":"Pinto","year":"2019","journal-title":"Mar. Environ. Res."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Silva, M., Rodr\u00edguez, I., Barreiro, A., Kaufmann, M., Neto, A.I., Hassouani, M., Sabour, B., Alfonso, A., Botana, L.M., and Vasconcelos, V. (2019). Tetrodotoxins occurrence in non-traditional vectors of the North Atlantic waters (Portuguese maritime territory, and Morocco coast). Toxins, 11.","DOI":"10.3390\/toxins11060306"},{"key":"ref_25","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_26","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_27","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1006\/abio.2000.4953","article-title":"Electrospray ionization mass spectrometry of tetrodotoxin and its analogs: Liquid chromatography\/mass spectrometry, tandem mass spectrometry, and liquid chromatography\/tandem mass spectrometry","volume":"290","author":"Shoji","year":"2001","journal-title":"Anal. Biochem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1468","DOI":"10.1080\/19440049.2016.1218070","article-title":"High-resolution mass spectrometry analysis of tetrodotoxin (TTX) and its analogues in puffer fish and shellfish","volume":"33","author":"Bane","year":"2016","journal-title":"Food Addit. Contam. A"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"142","DOI":"10.1016\/j.ab.2006.02.010","article-title":"Hydrophilic interaction liquid chromatography-electrospray ionization mass spectrometry of tetrodotoxin and its analogs","volume":"352","author":"Nakagawa","year":"2006","journal-title":"Anal. Biochem."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2799","DOI":"10.3390\/md11082799","article-title":"First identification of 5, 11-dideoxytetrodotoxin in marine animals, and characterization of major fragment ions of tetrodotoxin and its analogs by high resolution ESI-MS\/MS","volume":"11","author":"Abe","year":"2013","journal-title":"Mar. Drugs"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.chroma.2015.01.086","article-title":"Development of a sensitive and selective liquid chromatography-mass spectrometry method for high throughput analysis of paralytic shellfish toxins using graphitised carbon solid phase extraction","volume":"1387","author":"Boundy","year":"2015","journal-title":"J. Chromatogr. A"},{"key":"ref_32","unstructured":"European Commission (2002). Commission Decision of 15 March 2002 establishing special health checks for the harvesting and processing of certain bivalve mollusks with a level of amnesic shellfish poison (ASP) exceeding the limit laid down by Council Directive 91\/492\/EEC. Off. J. Eur. Comm., L 75, 65\u201366."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2621","DOI":"10.1080\/00222933.2012.724721","article-title":"Foregut anatomy and predation by Charonia lampas (gastropoda: Prosobranchia: Neotaenioglossa) attacking Ophidiaster ophidianus (asteroidea: Ophidiasteridae) in the Azores, with a review of triton feeding behavior","volume":"46","author":"Morton","year":"2012","journal-title":"J. Nat. Hist."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"617","DOI":"10.2331\/suisan.53.617","article-title":"Vibrio alginolyticus, a TTX-producing bacterium isolated from the starfish Astropecten polyacanthus","volume":"53","author":"Narita","year":"1987","journal-title":"Nippon Suisan Gakkaishi"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"573","DOI":"10.1016\/S0041-0101(00)00171-9","article-title":"Possible source of tetrodotoxin in the starfiish Astropecten scoparius","volume":"39","author":"Lin","year":"2001","journal-title":"Toxicon"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1016\/j.foodcont.2016.12.038","article-title":"Relevance and challenges in monitoring marine biotoxins in non-bivalve vectors","volume":"76","author":"Costa","year":"2017","journal-title":"Food Control"},{"key":"ref_37","unstructured":"EULRMB (2021, January 12). Determination of Tetrodontoxin byHILIC-MS\/MS, European Reference Laboratory for Marine Biotoxins, Ed 01062107. Available online: http:\/\/www.aecosan.msssi.gob.es\/en\/CRLMB\/docs\/docs\/metodos_analiticos_de_desarrollo\/HILIC-LCMSMS_SOP_for_TTX_in_mussels.pdf."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Turner, A.D., Dhanji-Rapkova, M., Coates, L., Bickerstaff, L., Milligan, S., O\u2019Neill, A., Faulkner, D., McEneny, H., Baker-Austin, C., and Lees, D.N. (2017). Detection of tetrodotoxin shellfish poisoning (tsp) toxins and causative factors in bivalve molluscs from the UK. Mar. Drugs, 15.","DOI":"10.3390\/md15090277"},{"key":"ref_39","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_40","doi-asserted-by":"crossref","first-page":"521","DOI":"10.1093\/jaoac\/78.2.521","article-title":"Detection of sodium channel toxins: Directed cytotoxicity assays for purified ciguatoxins, brevetoxins, saxitoxins and seafood extracts","volume":"78","author":"Manger","year":"1995","journal-title":"J. AOAC Int."}],"container-title":["Toxins"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6651\/13\/4\/250\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,13]],"date-time":"2025-10-13T13:25:37Z","timestamp":1760361937000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6651\/13\/4\/250"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,3,31]]},"references-count":42,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2021,4]]}},"alternative-id":["toxins13040250"],"URL":"https:\/\/doi.org\/10.3390\/toxins13040250","relation":{},"ISSN":["2072-6651"],"issn-type":[{"value":"2072-6651","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,3,31]]}}}