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Development and Validation of an LC-MS/MS Method for the Ultra-Trace Analysis of Pacific Ciguatoxins in Fish

BACKGROUND: Ciguatera fish poisoning (CFP) poses a serious threat to both public health and the use of aquatic resources from the various warm‐water regions of the world. Hence, a process for the efficient determination of the relevant toxins is required. OBJECTIVE: We sought to develop and validate...

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Autores principales: Nagae, Mika, Igarashi, Tomoji, Mizukoshi, Kazushi, Kuniyoshi, Kyoko, Oshiro, Naomasa, Yasumoto, Takeshi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8475085/
https://www.ncbi.nlm.nih.gov/pubmed/33831184
http://dx.doi.org/10.1093/jaoacint/qsab052
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author Nagae, Mika
Igarashi, Tomoji
Mizukoshi, Kazushi
Kuniyoshi, Kyoko
Oshiro, Naomasa
Yasumoto, Takeshi
author_facet Nagae, Mika
Igarashi, Tomoji
Mizukoshi, Kazushi
Kuniyoshi, Kyoko
Oshiro, Naomasa
Yasumoto, Takeshi
author_sort Nagae, Mika
collection PubMed
description BACKGROUND: Ciguatera fish poisoning (CFP) poses a serious threat to both public health and the use of aquatic resources from the various warm‐water regions of the world. Hence, a process for the efficient determination of the relevant toxins is required. OBJECTIVE: We sought to develop and validate the first LC-MS/MS method to quantify the major toxins prevalent in fish from the Pacific Ocean. METHOD: Toxins were extracted from fish flesh (2 g) using a methanol–water mixture (9:1, v/v). The extract was heated at 80°C, and low-polarity lipids were eliminated using hexane, initially from the basic solution and later from the acidic solution. The cleanup was performed using solid-phase extraction, Florisil, silica, reversed-phase C18, and primary secondary amine columns. A validation study was conducted by spiking fish flesh with two representative toxins having different skeletal structures and polarities and was calibrated by NMR (qNMR) spectroscopy. RESULTS: The validation parameters for the ciguatera toxins CTX1B and CTX3C at spiked levels of 0.1 µg/kg were as follows: repeatabilities of 2.3–3.5% and 3.2–5.3%; intermediate precisions of 6.3–9.8% and 6.0–7.4%; recoveries of 80–107% and 95–120%, respectively. The lowest detection levels were 0.004 µg/kg for CTX1B, 0.005 µg/kg for 51-hydroxyCTX3C, and 0.009 µg/kg for CTX3C. CONCLUSIONS: The described method practically clears the international action level of 0.01 µg/kg CTX1B equivalents set by the U.S. Food and Drug Administration and the European Food Safety Authority and satisfies the global standards set by Codex and AOAC INTERNATIONAL. HIGHLIGHTS: A validation study for an LC-MS/MS method for ciguatoxin detection was completed for the first time using calibrated toxin standards.
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spelling pubmed-84750852021-12-28 Development and Validation of an LC-MS/MS Method for the Ultra-Trace Analysis of Pacific Ciguatoxins in Fish Nagae, Mika Igarashi, Tomoji Mizukoshi, Kazushi Kuniyoshi, Kyoko Oshiro, Naomasa Yasumoto, Takeshi J AOAC Int Food Chemical Contaminants BACKGROUND: Ciguatera fish poisoning (CFP) poses a serious threat to both public health and the use of aquatic resources from the various warm‐water regions of the world. Hence, a process for the efficient determination of the relevant toxins is required. OBJECTIVE: We sought to develop and validate the first LC-MS/MS method to quantify the major toxins prevalent in fish from the Pacific Ocean. METHOD: Toxins were extracted from fish flesh (2 g) using a methanol–water mixture (9:1, v/v). The extract was heated at 80°C, and low-polarity lipids were eliminated using hexane, initially from the basic solution and later from the acidic solution. The cleanup was performed using solid-phase extraction, Florisil, silica, reversed-phase C18, and primary secondary amine columns. A validation study was conducted by spiking fish flesh with two representative toxins having different skeletal structures and polarities and was calibrated by NMR (qNMR) spectroscopy. RESULTS: The validation parameters for the ciguatera toxins CTX1B and CTX3C at spiked levels of 0.1 µg/kg were as follows: repeatabilities of 2.3–3.5% and 3.2–5.3%; intermediate precisions of 6.3–9.8% and 6.0–7.4%; recoveries of 80–107% and 95–120%, respectively. The lowest detection levels were 0.004 µg/kg for CTX1B, 0.005 µg/kg for 51-hydroxyCTX3C, and 0.009 µg/kg for CTX3C. CONCLUSIONS: The described method practically clears the international action level of 0.01 µg/kg CTX1B equivalents set by the U.S. Food and Drug Administration and the European Food Safety Authority and satisfies the global standards set by Codex and AOAC INTERNATIONAL. HIGHLIGHTS: A validation study for an LC-MS/MS method for ciguatoxin detection was completed for the first time using calibrated toxin standards. Oxford University Press 2021-04-08 /pmc/articles/PMC8475085/ /pubmed/33831184 http://dx.doi.org/10.1093/jaoacint/qsab052 Text en © AOAC INTERNATIONAL 2021. https://creativecommons.org/licenses/by-nc-nd/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs licence (http://creativecommons.org/licenses/by-nc-nd/4.0/ (https://creativecommons.org/licenses/by-nc-nd/4.0/) ), which permits non-commercial reproduction and distribution of the work, in any medium, provided the original work is not altered or transformed in any way, and that the work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Food Chemical Contaminants
Nagae, Mika
Igarashi, Tomoji
Mizukoshi, Kazushi
Kuniyoshi, Kyoko
Oshiro, Naomasa
Yasumoto, Takeshi
Development and Validation of an LC-MS/MS Method for the Ultra-Trace Analysis of Pacific Ciguatoxins in Fish
title Development and Validation of an LC-MS/MS Method for the Ultra-Trace Analysis of Pacific Ciguatoxins in Fish
title_full Development and Validation of an LC-MS/MS Method for the Ultra-Trace Analysis of Pacific Ciguatoxins in Fish
title_fullStr Development and Validation of an LC-MS/MS Method for the Ultra-Trace Analysis of Pacific Ciguatoxins in Fish
title_full_unstemmed Development and Validation of an LC-MS/MS Method for the Ultra-Trace Analysis of Pacific Ciguatoxins in Fish
title_short Development and Validation of an LC-MS/MS Method for the Ultra-Trace Analysis of Pacific Ciguatoxins in Fish
title_sort development and validation of an lc-ms/ms method for the ultra-trace analysis of pacific ciguatoxins in fish
topic Food Chemical Contaminants
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8475085/
https://www.ncbi.nlm.nih.gov/pubmed/33831184
http://dx.doi.org/10.1093/jaoacint/qsab052
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