Improved Isolation Procedures for Okadaic Acid Group Toxins from Shellfish (Mytilus edulis) and Microalgae (Prorocentrum lima).
Jane KilcoyneStephen BurrellCíara NultyRafael SalasElliott J WrightIsabelle RajotteChristopher Owen MilesPublished in: Marine drugs (2020)
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 (M. edulis), 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 Prorocentrum lima, that produced OA, DTX1, and their esters, over ~1.5 years with maximum cell densities of ~70,000 cells mL-1 observed. Toxin accumulation progressively increased over the study period, to ~0.7 and 2.1 mg L-1 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.
Keyphrases
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- single cell
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- induced apoptosis
- minimally invasive
- simultaneous determination
- mass spectrometry
- cell cycle arrest
- anaerobic digestion
- transcription factor
- stem cells
- healthcare
- oxidative stress
- high resolution mass spectrometry
- recombinant human
- cell proliferation
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- signaling pathway
- health information
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- high resolution
- mesenchymal stem cells
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- tandem mass spectrometry
- drug discovery