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Preparation of (18)O-labelled azaspiracids for accurate quantitation using liquid chromatography–mass spectrometry

Azaspiracids (AZAs) are a group of polyether marine algal toxins known to accumulate in shellfish, posing a risk to human health and the seafood industry. Analysis of AZAs is typically performed using LC–MS, which can suffer from matrix effects that significantly impact the accuracy of measurement r...

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Autores principales: Wright, Elliott J., Meija, Juris, McCarron, Pearse, Miles, Christopher O.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10556123/
https://www.ncbi.nlm.nih.gov/pubmed/37530793
http://dx.doi.org/10.1007/s00216-023-04868-4
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author Wright, Elliott J.
Meija, Juris
McCarron, Pearse
Miles, Christopher O.
author_facet Wright, Elliott J.
Meija, Juris
McCarron, Pearse
Miles, Christopher O.
author_sort Wright, Elliott J.
collection PubMed
description Azaspiracids (AZAs) are a group of polyether marine algal toxins known to accumulate in shellfish, posing a risk to human health and the seafood industry. Analysis of AZAs is typically performed using LC–MS, which can suffer from matrix effects that significantly impact the accuracy of measurement results. While the use of isotopic internal standards is an effective approach to correct for these effects, isotopically labelled standards for AZAs are not currently available. In this study, (18)O-labelled AZA1, AZA2, and AZA3 were prepared by reaction with H(2)(18)O under acidic conditions, and the reaction kinetics and sites of incorporation were studied using LC–HRMS/MS aided by mathematical analysis of their isotope patterns. Analysis of the isotopic incorporation in AZA1 and AZA3 indicated the presence of four exchangeable oxygen atoms. Excessive isomerization occurred during preparation of (18)O-labelled AZA2, suggesting a role for the 8-methyl group in the thermodynamic stability of AZAs. Neutralized mixtures of (18)O-labelled AZA1 and AZA3 were found to maintain their isotopic and isomeric integrities when stored at −20 °C and were used to develop an isotope-dilution LC–MS method which was applied to reference materials of shellfish matrices containing AZAs, demonstrating high accuracy and excellent reproducibility. Preparation of isotopically labelled compounds using the isotopic exchange method, combined with the kinetic analysis, offers a feasible way to obtain isotopically labelled internal standards for a wide variety of biomolecules to support reliable quantitation. GRAPHICAL ABSTRACT: [Image: see text] SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00216-023-04868-4.
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spelling pubmed-105561232023-10-07 Preparation of (18)O-labelled azaspiracids for accurate quantitation using liquid chromatography–mass spectrometry Wright, Elliott J. Meija, Juris McCarron, Pearse Miles, Christopher O. Anal Bioanal Chem Research Paper Azaspiracids (AZAs) are a group of polyether marine algal toxins known to accumulate in shellfish, posing a risk to human health and the seafood industry. Analysis of AZAs is typically performed using LC–MS, which can suffer from matrix effects that significantly impact the accuracy of measurement results. While the use of isotopic internal standards is an effective approach to correct for these effects, isotopically labelled standards for AZAs are not currently available. In this study, (18)O-labelled AZA1, AZA2, and AZA3 were prepared by reaction with H(2)(18)O under acidic conditions, and the reaction kinetics and sites of incorporation were studied using LC–HRMS/MS aided by mathematical analysis of their isotope patterns. Analysis of the isotopic incorporation in AZA1 and AZA3 indicated the presence of four exchangeable oxygen atoms. Excessive isomerization occurred during preparation of (18)O-labelled AZA2, suggesting a role for the 8-methyl group in the thermodynamic stability of AZAs. Neutralized mixtures of (18)O-labelled AZA1 and AZA3 were found to maintain their isotopic and isomeric integrities when stored at −20 °C and were used to develop an isotope-dilution LC–MS method which was applied to reference materials of shellfish matrices containing AZAs, demonstrating high accuracy and excellent reproducibility. Preparation of isotopically labelled compounds using the isotopic exchange method, combined with the kinetic analysis, offers a feasible way to obtain isotopically labelled internal standards for a wide variety of biomolecules to support reliable quantitation. GRAPHICAL ABSTRACT: [Image: see text] SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00216-023-04868-4. Springer Berlin Heidelberg 2023-08-02 2023 /pmc/articles/PMC10556123/ /pubmed/37530793 http://dx.doi.org/10.1007/s00216-023-04868-4 Text en © Crown 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Research Paper
Wright, Elliott J.
Meija, Juris
McCarron, Pearse
Miles, Christopher O.
Preparation of (18)O-labelled azaspiracids for accurate quantitation using liquid chromatography–mass spectrometry
title Preparation of (18)O-labelled azaspiracids for accurate quantitation using liquid chromatography–mass spectrometry
title_full Preparation of (18)O-labelled azaspiracids for accurate quantitation using liquid chromatography–mass spectrometry
title_fullStr Preparation of (18)O-labelled azaspiracids for accurate quantitation using liquid chromatography–mass spectrometry
title_full_unstemmed Preparation of (18)O-labelled azaspiracids for accurate quantitation using liquid chromatography–mass spectrometry
title_short Preparation of (18)O-labelled azaspiracids for accurate quantitation using liquid chromatography–mass spectrometry
title_sort preparation of (18)o-labelled azaspiracids for accurate quantitation using liquid chromatography–mass spectrometry
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10556123/
https://www.ncbi.nlm.nih.gov/pubmed/37530793
http://dx.doi.org/10.1007/s00216-023-04868-4
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