Tetrodotoxin (TTX) is one of the most potent toxins already isolated, which occurs in a wide variety of animals. In this work, the occurrence of TTX and analogues was examined using mass spectrometry, confocal microscopy, liquid chromatography-mass spectrometry (LC-MS), and mouse bioassay in a trumpet shell (Charonia lampas lampas) and in the fluids of a patient poisoned by consuming this shell. Retention time data in the LC-MS system within the enhanced mass spectrum (EMS) mode indicated the presence of TTX and the analogue 5,6,11-trideoxyTTX; the enhanced product ion (EPI) mode confirmed the existence of both toxins with the formation of characteristic daughter ions from the fragment pattern of each molecule. TTX and 5,6,11-trideoxyTTX were only detected in the digestive gland of the trumpet shell and also in the urine and serum of the patient. The concentration of 5,6,11-trideoxyTTX checked in the samples by LC-MS was 3 times higher than TTX. However, the results obtained by mouse bioassay showed that the analogue is much less toxic than TTX. In vitro toxicity was checked using cerebellar cells; in these experiments the trumpet shell sample showed high toxicity, but the level was lower than in vivo results probably due to some competition between analogues. This paper shows for first time the presence and toxicity of TTX and 5,6,11-trideoxyTTX in a trumpet shell collected in the European coasts. The LC-MS method is a useful tool to confirm the presence of TTX and the further identification of TTX analogues.
Ciguatera fish poisoning (CFP) is a human foodborne intoxication caused by ingestion of tropical fishes contaminated with the potent polyether toxins known as ciguatoxins (CTXs). These toxins are issued from Gambierdiscus species of dinoflagellates. Herbivorous fish accumulate these toxins in their musculature and viscera after ingesting dinoflagellates. Epidemiological studies showed that CFP has been present in areas between 35 degrees North and 35 degrees South latitude, mainly, Indo-pacific and Caribbean areas, but not in waters closed to European and African continent. In the present paper, a specimen of Seriola dumerili weighing 70 kg and a smaller Seriola fasciata specimen, captured in waters belonging to Selvagens Islands (Madeira Arquipelago), were analyzed. Fishes from this genus were implicated in previous suspected ciguatera poisoning outbreaks in the Portuguese Madeira Arquipelago in the North Atlantic Ocean. Analysis was performed by two approaches, a functional method using cerebellar granule cells and by ultraperformance liquid chromatography-mass spectrometry (UPLC-MS) method. The study was carried out in one portion of the tail muscle of Seriola fasciata and five parts of the body of Seriola dumerili (tail muscle, head, ventral muscle, mid muscle, and liver). The functional method consisted in the modification of the inward sodium current in cerebellar granule cells and the chemical method was a high resolution chromatography, which allowed elucidating the toxin profile in the samples. In addition, UPLC-MS technique was optimized and used for detecting and quantifying CTXs for the first time. After fish extraction and clean up, the chromatograms revealed the presence of CTX-1B at 1111.6 m/z, CTX-3C at 1023.5 m/z, a CTX analogue at 1040.6 m/z, and a CTX from the Caribbean or Indic waters at 1141.6 m/z. Therefore, the results obtained in the present paper for both methods confirm, for the first time, the presence of CTX in fish from Madeira Arquipelago.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.