Optimization of the Extraction Protocol for Pacific Ciguatoxins from Marine Products Prior to Analysis Using the Neuroblastoma Cell-Based Assay
Ciguatera poisoning (CP) is caused by the consumption of marine products contaminated with ciguatoxins (CTXs) produced by dinoflagellates of the genus <i>Gambierdiscus</i>. Analytical methods for CTXs, involving the extraction/purification of trace quantities of CTXs from complex matrice...
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2025-01-01
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| author | Thomas Yon Philippe Cruchet Jérôme Viallon J. Sam Murray Emillie Passfield Mireille Chinain Hélène Taiana Darius Mélanie Roué |
| author_facet | Thomas Yon Philippe Cruchet Jérôme Viallon J. Sam Murray Emillie Passfield Mireille Chinain Hélène Taiana Darius Mélanie Roué |
| author_sort | Thomas Yon |
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| description | Ciguatera poisoning (CP) is caused by the consumption of marine products contaminated with ciguatoxins (CTXs) produced by dinoflagellates of the genus <i>Gambierdiscus</i>. Analytical methods for CTXs, involving the extraction/purification of trace quantities of CTXs from complex matrices, are numerous in the literature. However, little information on their effectiveness for nonpolar CTXs is available, yet these congeners, contributing to the risk of CP, are required for the establishment of effective food safety monitoring programs. An evaluation of six extraction/purification protocols, performed with CTX3C spiked on fish flesh and a neuroblastoma cell-based assay (CBA-N2a), revealed recoveries from 6 to 45%. This led to the development of an optimized 3-day protocol designed for a large number of samples, with CTX1B and CTX3C eluting in a single fraction and showing recoveries of 73% and 70%, respectively. In addition, a reduction in adverse matrix effects in the CBA-N2a analyses was demonstrated with naturally contaminated specimens, increasing the sensitivity of the method, which now meets the very low guidance level recommended by international agencies. However, efforts are still required to reduce the signal suppression observed in LC-MS/MS analysis. This optimized protocol contributes to the technological advancement of detection methods, promoting food safety and improving CP risk assessment in marine products. |
| format | Article |
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| institution | Kabale University |
| issn | 1660-3397 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | MDPI AG |
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| series | Marine Drugs |
| spelling | doaj-art-ce7a07e4a6d44284b302c923a1001aea2025-01-24T13:39:36ZengMDPI AGMarine Drugs1660-33972025-01-012314210.3390/md23010042Optimization of the Extraction Protocol for Pacific Ciguatoxins from Marine Products Prior to Analysis Using the Neuroblastoma Cell-Based AssayThomas Yon0Philippe Cruchet1Jérôme Viallon2J. Sam Murray3Emillie Passfield4Mireille Chinain5Hélène Taiana Darius6Mélanie Roué7Institut Louis Malardé (ILM), UMR 241-SECOPOL (IFREMER, ILM, IRD, UPF), Laboratory of Marine Biotoxins, P.O. Box 30, 98713 Papeete, Tahiti, French PolynesiaInstitut Louis Malardé (ILM), UMR 241-SECOPOL (IFREMER, ILM, IRD, UPF), Laboratory of Marine Biotoxins, P.O. Box 30, 98713 Papeete, Tahiti, French PolynesiaInstitut Louis Malardé (ILM), UMR 241-SECOPOL (IFREMER, ILM, IRD, UPF), Laboratory of Marine Biotoxins, P.O. Box 30, 98713 Papeete, Tahiti, French PolynesiaCawthron Institute, 98 Halifax St East, PB 2, The Wood, Nelson 7010, New ZealandCawthron Institute, 98 Halifax St East, PB 2, The Wood, Nelson 7010, New ZealandInstitut Louis Malardé (ILM), UMR 241-SECOPOL (IFREMER, ILM, IRD, UPF), Laboratory of Marine Biotoxins, P.O. Box 30, 98713 Papeete, Tahiti, French PolynesiaInstitut Louis Malardé (ILM), UMR 241-SECOPOL (IFREMER, ILM, IRD, UPF), Laboratory of Marine Biotoxins, P.O. Box 30, 98713 Papeete, Tahiti, French PolynesiaInstitut de Recherche pour le Développement (IRD), UMR 241-SECOPOL (IFREMER, ILM, IRD, UPF), P.O. Box 6570, 98702 Faa’a, Tahiti, French PolynesiaCiguatera poisoning (CP) is caused by the consumption of marine products contaminated with ciguatoxins (CTXs) produced by dinoflagellates of the genus <i>Gambierdiscus</i>. Analytical methods for CTXs, involving the extraction/purification of trace quantities of CTXs from complex matrices, are numerous in the literature. However, little information on their effectiveness for nonpolar CTXs is available, yet these congeners, contributing to the risk of CP, are required for the establishment of effective food safety monitoring programs. An evaluation of six extraction/purification protocols, performed with CTX3C spiked on fish flesh and a neuroblastoma cell-based assay (CBA-N2a), revealed recoveries from 6 to 45%. This led to the development of an optimized 3-day protocol designed for a large number of samples, with CTX1B and CTX3C eluting in a single fraction and showing recoveries of 73% and 70%, respectively. In addition, a reduction in adverse matrix effects in the CBA-N2a analyses was demonstrated with naturally contaminated specimens, increasing the sensitivity of the method, which now meets the very low guidance level recommended by international agencies. However, efforts are still required to reduce the signal suppression observed in LC-MS/MS analysis. This optimized protocol contributes to the technological advancement of detection methods, promoting food safety and improving CP risk assessment in marine products.https://www.mdpi.com/1660-3397/23/1/42ciguatera poisoningciguatoxinsmarine productsextraction protocolCBA-N2aLC-MS/MS |
| spellingShingle | Thomas Yon Philippe Cruchet Jérôme Viallon J. Sam Murray Emillie Passfield Mireille Chinain Hélène Taiana Darius Mélanie Roué Optimization of the Extraction Protocol for Pacific Ciguatoxins from Marine Products Prior to Analysis Using the Neuroblastoma Cell-Based Assay Marine Drugs ciguatera poisoning ciguatoxins marine products extraction protocol CBA-N2a LC-MS/MS |
| title | Optimization of the Extraction Protocol for Pacific Ciguatoxins from Marine Products Prior to Analysis Using the Neuroblastoma Cell-Based Assay |
| title_full | Optimization of the Extraction Protocol for Pacific Ciguatoxins from Marine Products Prior to Analysis Using the Neuroblastoma Cell-Based Assay |
| title_fullStr | Optimization of the Extraction Protocol for Pacific Ciguatoxins from Marine Products Prior to Analysis Using the Neuroblastoma Cell-Based Assay |
| title_full_unstemmed | Optimization of the Extraction Protocol for Pacific Ciguatoxins from Marine Products Prior to Analysis Using the Neuroblastoma Cell-Based Assay |
| title_short | Optimization of the Extraction Protocol for Pacific Ciguatoxins from Marine Products Prior to Analysis Using the Neuroblastoma Cell-Based Assay |
| title_sort | optimization of the extraction protocol for pacific ciguatoxins from marine products prior to analysis using the neuroblastoma cell based assay |
| topic | ciguatera poisoning ciguatoxins marine products extraction protocol CBA-N2a LC-MS/MS |
| url | https://www.mdpi.com/1660-3397/23/1/42 |
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