{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,29]],"date-time":"2026-04-29T16:35:05Z","timestamp":1777480505485,"version":"3.51.4"},"reference-count":25,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2022,2,15]],"date-time":"2022-02-15T00:00:00Z","timestamp":1644883200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Marine Drugs"],"abstract":"<jats:p>The analysis of marine lipophilic toxins in shellfish products still represents a challenging task due to the complexity and diversity of the sample matrix. Liquid chromatography coupled with mass spectrometry (LC-MS) is the technique of choice for accurate quantitative measurements in complex samples. By combining unambiguous identification with the high selectivity of tandem MS, it provides the required high sensitivity and specificity. However, LC-MS is prone to matrix effects (ME) that need to be evaluated during the development and validation of methods. Furthermore, the large sample-to-sample variability, even between samples of the same species and geographic origin, needs a procedure to evaluate and control ME continuously. Here, we analyzed the toxins okadaic acid (OA), dinophysistoxins (DTX-1 and DTX-2), pectenotoxin (PTX-2), yessotoxin (YTX) and azaspiracid-1 (AZA-1). Samples were mussels (Mytilus galloprovincialis), both fresh and processed, and a toxin-free mussel reference material. We developed an accurate mass-extracted ion chromatogram (AM-XIC) based quantitation method using an Orbitrap instrument, evaluated the ME for different types and extracts of mussel samples, characterized the main compounds co-eluting with the targeted molecules and quantified toxins in samples by following a standard addition method (SAM). An AM-XIC based quantitation of lipophilic toxins in mussel samples using high resolution and accuracy full scan profiles (LC-HR-MS) is a good alternative to multi reaction monitoring (MRM) for instruments with HR capabilities. ME depend on the starting sample matrix and the sample preparation. ME are particularly strong for OA and related toxins, showing values below 50% for fresh mussel samples. Results for other toxins (AZA-1, YTX and PTX-2) are between 75% and 110%. ME in unknown matrices can be evaluated by comparing their full scan LC-HR-MS profiles with those of known samples with known ME. ME can be corrected by following SAM with AM-XIC quantitation if necessary.<\/jats:p>","DOI":"10.3390\/md20020143","type":"journal-article","created":{"date-parts":[[2022,2,15]],"date-time":"2022-02-15T22:44:47Z","timestamp":1644965087000},"page":"143","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["Quantitation Overcoming Matrix Effects of Lipophilic Toxins in Mytilus galloprovincialis by Liquid Chromatography-Full Scan High Resolution Mass Spectrometry Analysis (LC-HR-MS)"],"prefix":"10.3390","volume":"20","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8725-9100","authenticated-orcid":false,"given":"Camila Q. V.","family":"Costa","sequence":"first","affiliation":[{"name":"Centre of Marine Sciences (CCMAR\/CIMAR LA), University of Algarve, Campus de Gambelas, 8005-139 Faro, Portugal"}]},{"given":"In\u00eas I.","family":"Afonso","sequence":"additional","affiliation":[{"name":"Centre of Marine Sciences (CCMAR\/CIMAR LA), University of Algarve, Campus de Gambelas, 8005-139 Faro, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0167-7163","authenticated-orcid":false,"given":"Sandra","family":"Lage","sequence":"additional","affiliation":[{"name":"Centre of Marine Sciences (CCMAR\/CIMAR LA), University of Algarve, Campus de Gambelas, 8005-139 Faro, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6083-470X","authenticated-orcid":false,"given":"Pedro Reis","family":"Costa","sequence":"additional","affiliation":[{"name":"Centre of Marine Sciences (CCMAR\/CIMAR LA), University of Algarve, Campus de Gambelas, 8005-139 Faro, Portugal"},{"name":"Portuguese Institute for the Sea and Atmosphere (IPMA), Av. Bras\u00edlia, 1449-006 Lisbon, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6244-6468","authenticated-orcid":false,"given":"Adelino V. M.","family":"Can\u00e1rio","sequence":"additional","affiliation":[{"name":"Centre of Marine Sciences (CCMAR\/CIMAR LA), University of Algarve, Campus de Gambelas, 8005-139 Faro, Portugal"}]},{"given":"Jos\u00e9 P.","family":"Da Silva","sequence":"additional","affiliation":[{"name":"Centre of Marine Sciences (CCMAR\/CIMAR LA), University of Algarve, Campus de Gambelas, 8005-139 Faro, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2022,2,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Farabegoli, F., Blanco, L., Rodriguez, L., Vieites, J., and Cabado, A. (2018). Phycotoxins in Marine Shellfish: Origin, Occurrence and Effects on Humans. Mar. Drugs, 16.","DOI":"10.3390\/md16060188"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1051","DOI":"10.3389\/fmicb.2016.01051","article-title":"Marine Biotoxins: Occurrence, Toxicity, Regulatory Limits and Reference Methods","volume":"7","author":"Visciano","year":"2016","journal-title":"Front. Microbiol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"341","DOI":"10.1080\/02652030410001665919","article-title":"Distribution of diarrhetic shellfish poisoning toxins in consignments of blue mussel","volume":"21","author":"Jorgensen","year":"2004","journal-title":"Food Addit. Contam."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Klijnstra, M., Faassen, E., and Gerssen, A. (2021). A Generic LC-HRMS Screening Method for Marine and Freshwater Phycotoxins in Fish, Shellfish, Water, and Supplements. Toxins, 13.","DOI":"10.3390\/toxins13110823"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1371","DOI":"10.1093\/jaoac\/88.5.1371","article-title":"Liquid chromatography with mass spectrometry-Detection of lipophilic shellfish toxins","volume":"88","author":"Stobo","year":"2005","journal-title":"J. AOAC Int."},{"key":"ref_6","unstructured":"(2004). Regulation (EC) No 853\/2004 of the European parliament and of the council of 29 April 2004 laying down specific hygiene rules for food of animal origin. Off. J. Eur. Union, 139, 55\u2013205."},{"key":"ref_7","unstructured":"(2021). Commission Delegated Regulation (EU) 2021\/1374 of 12 April 2021 amending Annex III to Regulation (EC) No 853\/2004 of the European Parliament and of the Council on specific hygiene requirements for food of animal origin. Off. J. Eur. Union, 297, 1\u201315."},{"key":"ref_8","unstructured":"EURLMB (2022, January 29). EU-Harmonised Standard Operating Procedure for Determination of Lipophilic Marine Biotoxins in Molluscs by LC-MS\/MS, Version 5, Available online: https:\/\/www.aesan.gob.es\/CRLMB\/docs\/docs\/metodos_analiticos_de_desarrollo\/EU-Harmonised-SOP-LIPO-LCMSMS_Version5.pdf."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2939","DOI":"10.4155\/bio.12.243","article-title":"The future key role of LC-high-resolution-MS analyses in clinical laboratories: A focus on quantification","volume":"4","author":"Rochat","year":"2012","journal-title":"Bioanalysis"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"491","DOI":"10.1002\/mas.20298","article-title":"An Overview of Matrix Effects in Liquid Chromatography-Mass Spectrometry","volume":"30","author":"Trufelli","year":"2011","journal-title":"Mass Spectrom. Rev."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Cortese, M., Gigliobianco, M., Magnoni, F., Censi, R., and Di Martino, P. (2020). Compensate for or Minimize Matrix Effects? Strategies for Overcoming Matrix Effects in Liquid Chromatography-Mass Spectrometry Technique: A Tutorial Review. Molecules, 25.","DOI":"10.3390\/molecules25133047"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1024","DOI":"10.1080\/02652030802008601","article-title":"Approaches to the evaluation of matrix effects in the liquid chromatography-mass spectrometry (LC-MS) analysis of three regulated lipophilic toxin groups in mussel matrix (Mytilus edulis)","volume":"25","author":"Fux","year":"2008","journal-title":"Food Addit. Contam. Part A-Chem. Anal. Control. Expo. Risk Assess."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1213","DOI":"10.1007\/s00216-009-2790-0","article-title":"Solid phase extraction for removal of matrix effects in lipophilic marine toxin analysis by liquid chromatography-tandem mass spectrometry","volume":"394","author":"Gerssen","year":"2009","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1016\/j.toxicon.2020.10.002","article-title":"Evaluation of different strategies to minimize the matrix effects on LC-MS\/MS analysis of multiple lipophilic shellfish toxins in both acidic and alkaline chromatographic conditions","volume":"188","author":"Qiu","year":"2020","journal-title":"Toxicon"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"4529","DOI":"10.1016\/j.chroma.2009.03.062","article-title":"Sensitive method for the determination of lipophilic marine biotoxins in extracts of mussels and processed shellfish by high-performance liquid chromatography-tandem mass spectrometry based on enrichment by solid-phase extraction","volume":"1216","author":"These","year":"2009","journal-title":"J. Chromatogr. A"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2962","DOI":"10.1021\/acs.jafc.8b00346","article-title":"Selective Extraction and Purification of Azaspiracids from Blue Mussels (Mytilus edulis) Using Boric Acid Gel","volume":"66","author":"Miles","year":"2018","journal-title":"J. Agric. Food Chem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1093\/jat\/33.1.8","article-title":"Quantification of Saxitoxin and Neosaxitoxin in Human Urine Utilizing Isotope Dilution Tandem Mass Spectrometry","volume":"33","author":"Johnson","year":"2009","journal-title":"J. Anal. Toxicol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"273","DOI":"10.1016\/j.chroma.2007.05.016","article-title":"Development of an ultra-performance liquid chromatography-mass spectrometry method for the detection of lipophilic marine toxins","volume":"1157","author":"Fux","year":"2007","journal-title":"J. Chromatogr. A"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1016\/S0021-9673(01)01429-7","article-title":"Matrix effect and correction by standard addition in quantitative liquid chromatographic-mass spectrometric analysis of diarrhetic shellfish poisoning toxins","volume":"943","author":"Ito","year":"2002","journal-title":"J. Chromatogr. A"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"622","DOI":"10.1016\/j.talanta.2018.07.050","article-title":"A single run UPLC-MS\/MS method for detection of all EU-regulated marine toxins","volume":"189","author":"Rodriguez","year":"2018","journal-title":"Talanta"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"805","DOI":"10.1021\/ed076p805","article-title":"Estimates of precision in a standard additions analysis","volume":"76","author":"Bruce","year":"1999","journal-title":"J. Chem. Educ."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/j.chroma.2015.08.064","article-title":"High resolution mass spectrometry for quantitative analysis and untargeted screening of algal toxins in mussels and passive samplers","volume":"1416","author":"Zendong","year":"2015","journal-title":"J. Chromatogr. A"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"8522","DOI":"10.1021\/acs.est.6b02081","article-title":"Passive Sampling and High Resolution Mass Spectrometry for Chemical Profiling of French Coastal Areas with a Focus on Marine Biotoxins","volume":"50","author":"Zendong","year":"2016","journal-title":"Environ. Sci. Technol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"119077","DOI":"10.1016\/j.ijpharm.2020.119077","article-title":"Quantification of phospholipid degradation products in liposomal pharmaceutical formulations by ultra performance liquid chromatography-mass spectrometry (UPLC-MS)","volume":"578","author":"Siriwardane","year":"2020","journal-title":"Int. J. Pharm."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1453","DOI":"10.1007\/s00216-015-9245-6","article-title":"A study of inter-species ion suppression in electrospray ionization-mass spectrometry of some phospholipid classes","volume":"408","author":"Khoury","year":"2016","journal-title":"Anal. Bioanal. Chem."}],"container-title":["Marine Drugs"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1660-3397\/20\/2\/143\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:20:26Z","timestamp":1760134826000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1660-3397\/20\/2\/143"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,2,15]]},"references-count":25,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2022,2]]}},"alternative-id":["md20020143"],"URL":"https:\/\/doi.org\/10.3390\/md20020143","relation":{},"ISSN":["1660-3397"],"issn-type":[{"value":"1660-3397","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,2,15]]}}}