Improved Isolation Procedures for Okadaic Acid Group Toxins from Shellfish (<i>Mytilus edulis</i>) and Microalgae (<i>Prorocentrum lima</i>)
Okadaic acid (OA) group toxins may accumulate in shellfish and can result in diarrhetic shellfish poisoning when consumed by humans, and are therefore regulated. Purified toxins are required for the production of certified reference materials used to accurately quantitate toxin levels in shellfish a...
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| Format: | Article |
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MDPI AG
2020-12-01
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| Series: | Marine Drugs |
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| Online Access: | https://www.mdpi.com/1660-3397/18/12/647 |
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| author | Jane Kilcoyne Stephen Burrell Cíara Nulty Rafael Salas Elliott J. Wright Isabelle Rajotte Christopher O. Miles |
| author_facet | Jane Kilcoyne Stephen Burrell Cíara Nulty Rafael Salas Elliott J. Wright Isabelle Rajotte Christopher O. Miles |
| author_sort | Jane Kilcoyne |
| collection | DOAJ |
| description | Okadaic acid (OA) group toxins may accumulate in shellfish and can result in diarrhetic shellfish poisoning when consumed by humans, and are therefore regulated. Purified toxins are required for the production of certified reference materials used to accurately quantitate toxin levels in shellfish and water samples, and for other research purposes. An improved procedure was developed for the isolation of dinophysistoxin 2 (DTX2) from shellfish (<i>M. edulis</i>), reducing the number of purification steps from eight to five, thereby increasing recoveries to ~68%, compared to ~40% in a previously reported method, and a purity of >95%. Cell densities and toxin production were monitored in cultures of <i>Prorocentrum lima</i>, that produced OA, DTX1, and their esters, over ~1.5 years with maximum cell densities of ~70,000 cells mL<sup>−1</sup> observed. Toxin accumulation progressively increased over the study period, to ~0.7 and 2.1 mg L<sup>−1</sup> of OA and DTX1 (including their esters), respectively, providing information on appropriate harvesting times. A procedure for the purification of OA and DTX1 from the harvested biomass was developed employing four purification steps, with recoveries of ~76% and purities of >95% being achieved. Purities were confirmed by LC-HRMS, LC-UV, and NMR spectroscopy. Additional stability observations led to a better understanding of the chemistry of these toxins. |
| first_indexed | 2024-03-10T14:02:35Z |
| format | Article |
| id | doaj.art-ff812dc0c5324d37ac0cecf625d8f139 |
| institution | Directory Open Access Journal |
| issn | 1660-3397 |
| language | English |
| last_indexed | 2024-03-10T14:02:35Z |
| publishDate | 2020-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Marine Drugs |
| spelling | doaj.art-ff812dc0c5324d37ac0cecf625d8f1392023-11-21T01:03:21ZengMDPI AGMarine Drugs1660-33972020-12-01181264710.3390/md18120647Improved Isolation Procedures for Okadaic Acid Group Toxins from Shellfish (<i>Mytilus edulis</i>) and Microalgae (<i>Prorocentrum lima</i>)Jane Kilcoyne0Stephen Burrell1Cíara Nulty2Rafael Salas3Elliott J. Wright4Isabelle Rajotte5Christopher O. Miles6Marine Institute, Rinville, Oranmore, Co. Galway H91 R673, IrelandMarine Institute, Rinville, Oranmore, Co. Galway H91 R673, IrelandMarine Institute, Rinville, Oranmore, Co. Galway H91 R673, IrelandMarine Institute, Rinville, Oranmore, Co. Galway H91 R673, IrelandBiotoxin Metrology, National Research Council Canada, Halifax, NS B3H 3Z1, CanadaBiotoxin Metrology, National Research Council Canada, Halifax, NS B3H 3Z1, CanadaBiotoxin Metrology, National Research Council Canada, Halifax, NS B3H 3Z1, CanadaOkadaic acid (OA) group toxins may accumulate in shellfish and can result in diarrhetic shellfish poisoning when consumed by humans, and are therefore regulated. Purified toxins are required for the production of certified reference materials used to accurately quantitate toxin levels in shellfish and water samples, and for other research purposes. An improved procedure was developed for the isolation of dinophysistoxin 2 (DTX2) from shellfish (<i>M. edulis</i>), reducing the number of purification steps from eight to five, thereby increasing recoveries to ~68%, compared to ~40% in a previously reported method, and a purity of >95%. Cell densities and toxin production were monitored in cultures of <i>Prorocentrum lima</i>, that produced OA, DTX1, and their esters, over ~1.5 years with maximum cell densities of ~70,000 cells mL<sup>−1</sup> observed. Toxin accumulation progressively increased over the study period, to ~0.7 and 2.1 mg L<sup>−1</sup> of OA and DTX1 (including their esters), respectively, providing information on appropriate harvesting times. A procedure for the purification of OA and DTX1 from the harvested biomass was developed employing four purification steps, with recoveries of ~76% and purities of >95% being achieved. Purities were confirmed by LC-HRMS, LC-UV, and NMR spectroscopy. Additional stability observations led to a better understanding of the chemistry of these toxins.https://www.mdpi.com/1660-3397/18/12/647OA group toxinsDSPpurificationshellfish<i>Prorocentrum lima</i>marine biotoxins |
| spellingShingle | Jane Kilcoyne Stephen Burrell Cíara Nulty Rafael Salas Elliott J. Wright Isabelle Rajotte Christopher O. Miles Improved Isolation Procedures for Okadaic Acid Group Toxins from Shellfish (<i>Mytilus edulis</i>) and Microalgae (<i>Prorocentrum lima</i>) Marine Drugs OA group toxins DSP purification shellfish <i>Prorocentrum lima</i> marine biotoxins |
| title | Improved Isolation Procedures for Okadaic Acid Group Toxins from Shellfish (<i>Mytilus edulis</i>) and Microalgae (<i>Prorocentrum lima</i>) |
| title_full | Improved Isolation Procedures for Okadaic Acid Group Toxins from Shellfish (<i>Mytilus edulis</i>) and Microalgae (<i>Prorocentrum lima</i>) |
| title_fullStr | Improved Isolation Procedures for Okadaic Acid Group Toxins from Shellfish (<i>Mytilus edulis</i>) and Microalgae (<i>Prorocentrum lima</i>) |
| title_full_unstemmed | Improved Isolation Procedures for Okadaic Acid Group Toxins from Shellfish (<i>Mytilus edulis</i>) and Microalgae (<i>Prorocentrum lima</i>) |
| title_short | Improved Isolation Procedures for Okadaic Acid Group Toxins from Shellfish (<i>Mytilus edulis</i>) and Microalgae (<i>Prorocentrum lima</i>) |
| title_sort | improved isolation procedures for okadaic acid group toxins from shellfish i mytilus edulis i and microalgae i prorocentrum lima i |
| topic | OA group toxins DSP purification shellfish <i>Prorocentrum lima</i> marine biotoxins |
| url | https://www.mdpi.com/1660-3397/18/12/647 |
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