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Effect of environmental and nutritional factors on growth and azaspiracid production of the dinoflagellate Azadinium spinosum ArchiMer
Jauffrais, Thierry; Sechet, Veronique; Herrenknecht, Christine; Truquet, Philippe; Savar, Veronique; Tillmann, Urban; Hess, Philipp.
Azadinium spinosum, a small dinoflagellate isolated from the North Sea, is a producer of azaspiracids (AZAs), a group of biotoxins associated with human illness following ingestion of contaminated shellfish. Using batch and continuous cultures of A. spinosum, the present study investigated the effects of different environmental and nutritional factors (salinity, temperature, photon flux density, aeration, culture media, nitrogen sources, phosphate source, and N/P ratios) on growth, maximum cell concentration, and AZA cell quota. Azadinium spinosum grew in a wide range of conditions; from 10 ̊C to 26 ̊C and salinities from 30 to 40, under irradiances ranging from 50 μmol m−2 s−1 to 250 μmol m−2 s−1, with or without aeration. Growth and maximum cell...
Tipo: Text Palavras-chave: AZA; Chemostat; Dinophyceae; Nutrients; N/P ratio; Photobioreactor.
Ano: 2013 URL: http://archimer.ifremer.fr/doc/00149/26054/24411.pdf
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Cytotoxicity, Fractionation and Dereplication of Extracts of the Dinoflagellate Vulcanodinium rugosum, a Producer of Pinnatoxin G ArchiMer
Geiger, Marie; Desanglois, Gwenaelle; Hogeveen, Kevin; Fessard, Valerie; Lepretre, Thomas; Mondeguer, Florence; Guitton, Yann; Herve, Fabienne; Sechet, Veronique; Grovel, Olivier; Pouchus, Yves-francois; Hess, Philipp.
Pinnatoxin G (PnTX-G) is a marine toxin belonging to the class of cyclic imines and produced by the dinoflagellate Vulcanodinium rugosum. In spite of its strong toxicity to mice, leading to the classification of pinnatoxins into the class of “fast-acting toxins”, its hazard for human health has never been demonstrated. In this study, crude extracts of V. rugosum exhibited significant cytotoxicity against Neuro2A and KB cells. IC50 values of 0.38 µg mL−1 and 0.19 µg mL−1 were estimated on Neuro2A cells after only 24 h of incubation and on KB cells after 72 h of incubation, respectively. In the case of Caco-2 cells 48 h after exposure, the crude extract of V. rugosum induced cell cycle arrest accompanied by a dramatic increase in double strand DNA breaks,...
Tipo: Text Palavras-chave: Dereplication; Cyclic imine; HRMS; Bioactivity; Pinnatoxins.
Ano: 2013 URL: http://archimer.ifremer.fr/doc/00156/26723/24802.pdf
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Extended evaluation of polymeric and lipophilic sorbents for passive sampling of marine toxins ArchiMer
Zendong, Zita; Herrenknecht, Christine; Abadie, Eric; Brissard, Charline; Tixier, Celine; Mondeguer, Florence; Sechet, Veronique; Amzil, Zouher; Hess, Philipp.
Marine biotoxins are algal metabolites that can accumulate in fish or shellfish and render these foodstuffs unfit for human consumption. These toxins, released into seawater during algal occurrences, can be monitored through passive sampling. Acetone, methanol and isopropanol were evaluated for their efficiency in extracting toxins from algal biomass. Isopropanol was chosen for further experiments thanks to a slightly higher recovery and no artifact formation. Comparison of Oasis HLB, Strata-X, BondElut C18 and HP-20 sorbent materials in SPE-mode led to the choice of Oasis HLB, HP-20 and Strata-X. These three sorbents were separately exposed as passive samplers for 24 h to seawater spiked with algal extracts containing known amounts of okadaic acid (OA),...
Tipo: Text Palavras-chave: Passive sampling; Oasis HLB; Strata-X; LOPE; PDMS; SPATT.
Ano: 2014 URL: https://archimer.ifremer.fr/doc/00186/29756/28187.pdf
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A mussel (Mytilus edulis) tissue certified reference material for the marine biotoxins azaspiracids ArchiMer
Mccarron, Pearse; Giddings, Sabrina D.; Reeves, Kelley L.; Hess, Philipp; Quilliam, Michael A..
Azaspiracids (AZAs) are lipophilic biotoxins produced by marine algae that can contaminate shellfish and cause human illness. The European Union (EU) regulates the level of AZAs in shellfish destined for the commercial market, with liquid chromatography-mass spectrometry (LC-MS) being used as the official reference method for regulatory analysis. Certified reference materials (CRMs) are essential tools for the development, validation, and quality control of LC-MS methods. This paper describes the work that went into the planning, preparation, characterization, and certification of CRM-AZA-Mus, a tissue matrix CRM, which was prepared as a wet homogenate from mussels (Mytilus edulis) naturally contaminated with AZAs. The homogeneity and stability of...
Tipo: Text Palavras-chave: Azaspiracids; Seafood safety; Matrix certified reference material; Liquid chromatography-mass spectrometry; Matrix effects.
Ano: 2015 URL: http://archimer.ifremer.fr/doc/00320/43138/42761.pdf
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Field and mesocosm trials on passive sampling for the study of adsorption and desorption behaviour of lipophilic toxins with a focus on OA and DTX1 ArchiMer
Fux, E; Marcaillou-le Baut, Claire; Mondeguer, Florence; Bire, R; Hess, Philipp.
It has been demonstrated that polymeric resins can be used as receiving phase in passive samplers designed for the detection of lipophilic marine toxins at sea and was referred to as solid phase adsorption toxin tracking (SPATT). The present study describes the uptake and desorption behaviour of the lipophilic marine toxins okadaic acid (OA) and dinophysistoxin-1 (DTX1) from Prorocentrum lima cultures by five styrene-divinylbenzene based polymeric resins Sepabeads (R) SP850, Sepabeads (R) SP825L, Amberlite (R) XAD4, Dowex (R) Optipore (R) L-493 and Diaion (R) HP-20. All resins accumulated OA and DTX1 from the P. lima culture with differences in adsorption rate and equilibrium rate. Following statistical evaluation, HP-20, SP850 and SP825L demonstrated...
Tipo: Text Palavras-chave: Polymeric resin; Dinophysistoxin 1; Okadaic acid; SPATT; Prorocentrum lima.
Ano: 2008 URL: http://archimer.ifremer.fr/doc/2008/publication-4739.pdf
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Combined effects of temperature, irradiance and pH on Teleaulax amphioxeia (Cryptophyceae) physiology and feeding ratio for its predator Mesodinium rubrum (Ciliophora) ArchiMer
Gaillard, Sylvain; Charrier, Aurelie; Malo, Florent; Carpentier, Liliane; Bougaran, Gael; Hégaret, Helene; Réveillon, Damien; Hess, Philipp; Séchet, Veronique.
The cryptophyte Teleaulax amphioxeia is a source of plastids for the ciliate Mesodinium rubrum and both organisms are members of the trophic chain of several species of Dinophysis. It is important to better understand the ecology of organisms at the first trophic levels before assessing the impact of principal factors of global change on Dinophysis spp. Therefore, combined effects of temperature, irradiance and pH on growth rate, photosynthetic activity and pigment content of a temperate strain of T. amphioxeia were studied using a full factorial design (central composite design 23*) in 17 individually controlled bioreactors. The derived model predicted an optimal growth rate of T. amphioxeia at a light intensity of 400 µmol photons · m‐2 · s‐1, more...
Tipo: Text Palavras-chave: Dinophysis; Ecophysiology; Full factorial design; Global change; Mesodinium rubrum; Teleaulax amphioxeia.
Ano: 2020 URL: https://archimer.ifremer.fr/doc/00609/72155/70910.pdf
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Dissolved azaspiracids are absorbed and metabolized by blue mussels (Mytilus edulis) ArchiMer
Jauffrais, Thierry; Kilcoyne, Jane; Herrenknecht, Christine; Truquet, Philippe; Sechet, Veronique; Miles, Christopher O.; Hess, Philipp.
The relationship between azaspiracid shellfish poisoning and a small dinoflagellate, Azadinium spinosum, has been shown recently. The organism produces AZA1 and -2, while AZA3 and other analogues are metabolic products formed in shellfish. We evaluated whether mussels were capable of accumulating dissolved AZA1 and -2, and compared the toxin profiles of these mussels at 24 h with profiles of those exposed to live or lysed A. spinosum. We also assessed the possibility of preparative production of AZA metabolites by exposing mussels to semi-purified AZA1. We exposed mussels to similar concentration of AZAs: dissolved AZA1+2 (crude extract) at 7.5 and 0.75 µg L-1, dissolved AZA1+2 (7.5 µg L-1) in combination with Isochrysis affinis galbana, and lysed and live...
Tipo: Text Palavras-chave: Dissolved marine biotoxins; AZA; Tissue distribution; Bivalve molluscs; LC-MS/MS; Azaspiracid.
Ano: 2013 URL: http://archimer.ifremer.fr/doc/00136/24757/22867.pdf
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Effect of seawater salinity on pore-size distribution on a poly(styrene)-based HP20 resin and its adsorption of diarrhetic shellfish toxins ArchiMer
Fan, Lin; Sun, Geng; Qiu, Jiangbing; Ma, Qimin; Hess, Philipp; Li, Aifeng.
In the present study, okadaic acid (OA) and dinophysistoxin-1 (DTX1) were spiked into artificial seawater at low, medium and high estuarine salinities (9‰, 13.5‰ and 27‰). Passive samplers (HP20 resin) used for solid phase adsorption toxin tracking (SPATT) technology were exposed in these seawaters for12-h periods. Adsorption curves well fitted a pseudo-secondary kinetics model. The highest initial sorption rates of both toxins occurred in the seawater of medium salinity, followed by seawater of low and high estuarine salinity. Pore volumes of micropores (< 2 nm) and small mesopores (2 nm < diameter < 10 nm) of HP20 resin decreased after adsorption of toxins in seawater at high and low salinity but not in seawater at medium salinity, which...
Tipo: Text Palavras-chave: Diarrhetic shellfish toxins (DST); Solid phase adsorption toxin tracking (SPATT); HP20 resin; Salinity; Pore-size distribution; Pseudo-secondary kinetics equation.
Ano: 2014 URL: https://archimer.ifremer.fr/doc/00226/33766/32192.pdf
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Growth and toxin production of Azadinium spinosum in batch and continuous culture ArchiMer
Jauffrais, Thierry; Sechet, Veronique; Herrenknecht, Christine; Tillmann, Urban; Krock, Bernd; Amzil, Zouher; Hess, Philipp.
Azaspiracids are lipophilic marine biotoxins causing gastrointestinal symptoms similar to DSP toxins. Since 1995, azaspiracids have been encountered in Europe, Africa and more recently in North and South America and Japan. The biological primary producer remained undiscovered during many years and has now been identified as Azadinium spinosum. The organism was grown using K modified medium, at 18°C with a PFD of 200 μmol.m-2.s-1 and a photoperiod of 16L/8D. Batch cultures were carried out using 75mL and 10L flasks, while continuous cultures were produced in 100L chemostats. Cells were recovered using centrifugation or filtration. Different extraction solvents and procedures as well as evaporation modes were evaluated for yield. Quantitation was carried out...
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Ano: 2010 URL: http://archimer.ifremer.fr/doc/00152/26348/24431.pdf
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Algal toxin profiles in Nigerian coastal waters (Gulf of Guinea) using passive sampling and liquid chromatography coupled to mass spectrometry ArchiMer
Zendong, Suzie Zita; Kadiri, Medina; Herrenknecht, Christine; Nezan, Elisabeth; Mazzeo, Antonia; Hess, Philipp.
Algal toxins may accumulate in fish and shellfish and thus cause poisoning in consumers of seafood. Such toxins and the algae producing them are regularly surveyed in many countries, including Europe, North America, Japan and others. However, very little is known regards the occurrence of such algae and their toxins in most African countries. This paper reports on a survey of phytoplankton and algal toxins in Nigerian coastal waters. Seawater samples were obtained from four sites for phytoplankton identification, on three occasions between the middle of October 2014 and the end of February 2015 (Bar Beach and Lekki in Lagos State, Port Harcourt in Rivers State and Uyo in Akwa Ibom State). The phytoplankton community was generally dominated by diatoms and...
Tipo: Text Palavras-chave: Dinoflagellates; Dinophysis; Phycotoxins; Untargeted analysis; Phytoplankton.
Ano: 2016 URL: http://archimer.ifremer.fr/doc/00314/42521/41893.pdf
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Ostreopsis cf. ovata Bloom in Currais, Brazil: Phylogeny, Toxin Profile and Contamination of Mussels and Marine Plastic Litter ArchiMer
Tibiriçá, Carlos Eduardo J. A.; Leite, Isabel P.; Batista, Talita V. V.; Fernandes, Luciano F.; Chomérat, Nicolas; Herve, Fabienne; Hess, Philipp; Mafra, Luiz.
Ostreopsis cf. ovata is a toxic marine benthic dinoflagellate responsible for harmful blooms affecting ecosystem and human health, mostly in the Mediterranean Sea. In this study we report the occurrence of a summer O. cf. ovata bloom in Currais, a coastal archipelago located on the subtropical Brazilian coast (~25° S). This bloom was very similar to Mediterranean episodes in many aspects: (a) field-sampled and cultivated O. cf. ovata cells aligned phylogenetically (ITS and LSU regions) along with Mediterranean strains; (b) the bloom occurred at increasing temperature and irradiance, and decreasing wind speed; (c) cell densities reached up to 8.0 × 104 cell cm−2 on fiberglass screen and 5.6 × 105 cell g−1 fresh weight on seaweeds; (d) and toxin profiles...
Tipo: Text Palavras-chave: Harmful algal bloom; Benthic microalgae; Toxic dinoflagellates; Ovatoxin; Toxin transfer; Seafood safety; Marine pollution; Plastic litter; Biofilm formation.
Ano: 2019 URL: https://archimer.ifremer.fr/doc/00509/62037/66177.pdf
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Effect of Dilution Rate on Azadinium spinosum and Azaspiracid (AZA) Production in Pilot Scale Photobioreactors for the Harvest of AZA1 and -2 ArchiMer
Jauffrais, Thierry; Sechet, Veronique; Truquet, Philippe; Amzil, Zouher; Herrenknecht, Christine; Hess, Philipp.
Azadinium spinosum, a small dinoflagellate has recently been discovered and identified as the primary producer of azaspiracid-1 (AZA) and -2. Since AZA poisoning has been reported following consumption of contaminated shellfish it is important to have these toxins available for toxicological studies, and a sustainable production of AZAs as calibrants in monitoring programs without having to rely on natural events. In order to address this concern, continuous pilot scale cultures were carried out to evaluate the feasibility of AZA production from A. spinosum. Algae were cultured using two 100 L chemostats in series (R1 and R2), with agitation and pH control. Four different dilution rates were tested (0.15, 0.2, 0.25 and 0.3 day−1) to evaluate chemostat...
Tipo: Text Palavras-chave: Solid phase adsorption; Chemostat; Photobioreactor; Microalgae; LC-MS/MS; Tangential flow filtration.
Ano: 2014 URL: http://archimer.ifremer.fr/doc/00152/26349/24437.pdf
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Isolation, Structure Elucidation, Relative LC-MS Response, and in Vitro Toxicity of Azaspiracids from the Dinoflagellate Azadinium spinosum ArchiMer
Kilcoyne, Jane; Nulty, Ciara; Jauffrais, Thierry; Mccarron, Pearse; Herve, Fabienne; Foley, Barry; Rise, Frode; Crain, Sheila; Wilkins, Alistair L.; Twiner, Michael J.; Hess, Philipp; Miles, Christopher O..
We identified three new azaspiracids (AZAs) with molecular weights of 715, 815, and 829 (AZA33 (3), AZA34 (4), and AZA35, respectively) in mussels, seawater, and Azadinium spinosum culture. Approximately 700 mu g of 3 and 250 mu g of 4 were isolated from a bulk culture of A. spinosum, and their structures determined by MS and NMR spectroscopy. These compounds differ significantly at the carboxyl end of the molecule from known AZA analogues and therefore provide valuable information on structure-activity relationships. Initial toxicological assessment was performed using an in vitro model system based on Jurkat T lymphocyte cytotoxicity, and the potencies of 3 and 4 were found to be 0.22- and 5.5-fold that of AZA1 (1), respectively. Thus, major changes in...
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Ano: 2014 URL: https://archimer.ifremer.fr/doc/00245/35606/34414.pdf
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World Health Organization and international guidelines for toxin control, harmful algal bloom management, and response planning ArchiMer
Soltani, Alex; Hess, Philipp; Dixon, Mike B; Boerlage, Siobhan F.e.; Anderson, Donald M; Newcombe, Gayle; House, Jenny; Ho, Lionel; Baker, Peter; Burch, Michael.
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Ano: 2017 URL: http://archimer.ifremer.fr/doc/00407/51836/52437.pdf
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COLNACOQ. Composés lipophiles naturels dans l’environnement des coquillages. Rapport final ArchiMer
Hess, Philipp; Geiger, Marie; Brochard, Solène; Lepretre, Thomas; Fessard, Valerie; Antignac, Jean-philippe; Dupont, Jacques; Sechet, Veronique; Vanel, Faustine; Mondeguer, Florence; Herve, Fabienne; Leborgne, Sabrina; Deslanglois, Gwenaëlle; Marshall, Lindsey; Rounds, Lucy; Guitton, Yann; Amzil, Zouher; Grovel, Olivier; Ruiz, Nicolas; Pouchus, Francois.
Cette étude s’est inscrite dans un contexte d’évaluation de la salubrité des mollusques bivalves destinés à la consommation humaine. Ces organismes peuvent en effet accumuler des toxines, en particulier de microalgues, en concentrations importantes, pouvant induire des intoxications chez les personnes les consommant. Afin d’assurer la protection des consommateurs, les toxines réglementées sont recherchées dans les coquillages par techniques physico-chimiques. En parallèle, les toxines émergentes, potentiellement produites par d’autres organismes que les micro-algues telles que les micromycètes, peuvent être detectées par l’utilisation du test de toxicité aigüe sur souris. Cependant, ce bio-essai présente de très nombreux inconvénients, notamment éthiques...
Tipo: Text Palavras-chave: Cytotoxicité; Toxicité sur larves de diptères; Activité antibactérienne; Bactéries marines; Toxines lipophiles; Micro-algues; Dinoflagellés; Micromycètes; Penicillium; Acide okadaïque; Azaspiracide; Pinnatoxine G; Vulcanodinium rugosum; Prorocentrum lima; Azadinium spinosum; Beauveria brongniartii.
Ano: 2015 URL: http://archimer.ifremer.fr/doc/00253/36470/35015.pdf
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Stratégie de mesure non ciblée de type métabolomique (couplage LC-HRMS ) pour identifier différents groupes de composés bioactifs accumulés dans les mollusques bivalves ArchiMer
Mondeguer, Florence; Antignac, Jean-philippe; Guitton, Yann; Monteau, Fabrice; Le Borgne, Sabrina; Hess, Philipp.
La bioactivité des toxines du phytoplancton qui s’accumulent dans les coquillages est presque toujours évaluée sur modèle animal (test souris). Malgré ses avantages, la capacité de ce test à expliquer la nature de cette bio-activité reste limitée. Par ailleurs, le contrôle sanitaire actuellement basé sur une méthode ciblée d’identification et de quantification d’un ensemble de toxines connues ne permet pas de détecter des toxines encore inconnues. Afin de répondre à ce besoin de caractérisation de substances toxiques inconnues, une nouvelle approche de profilage chimique différentiel et non ciblé, de type métabolomique, a été proposée. Les 2 extraits sélectionnés sont ceux ayant montré une toxicité positive chez la souris, sans que les substances...
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Ano: 2013 URL: http://archimer.ifremer.fr/doc/00179/29042/27480.pdf
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Azaspiracid shellfish poisoning: A review on the chemistry, ecology, and toxicology with an emphasis on human health impacts ArchiMer
Twiner, Michael J.; Rehmann, Nils; Hess, Philipp; Doucette, Gregory J..
Azaspiracids (AZA) are polyether marine toxins that accumulate in various shellfish species and have been associated with severe gastrointestinal human intoxications since 1995. This toxin class has since been reported from several countries, including Morocco and much of western Europe. A regulatory limit of 160 mu g AZA/kg whole shellfish flesh was established by the EU in order to protect human health; however, in some cases, AZA concentrations far exceed the action level. Herein we discuss recent advances on the chemistry of various AZA analogs, review the ecology of AZAs, including the putative progenitor algal species, collectively interpret the in vitro and in vivo data on the toxicology of AZAs relating to human health issues, and outline the...
Tipo: Text Palavras-chave: Azaspiracid (AZA); AZP; Shellfish poisoning.
Ano: 2008 URL: http://archimer.ifremer.fr/doc/00077/18805/16375.pdf
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Chemically mediated interactions between Microcystis and Planktothrix: Impact on their physiology and metabolic profiles ArchiMer
Briand, Enora; Reubrech, Sébastien; Mondeguer, Florence; Sibat, Manoella; Hess, Philipp; Amzil, Zouher; Bormans, Myriam.
Freshwater cyanobacteria are well known for their ability to produce a wide variety of bioactive compounds, some of which have been described as allelochemicals. There is growing evidence that these secondary metabolites play an important role in shaping community composition through biotic interactions; however, for the most part, their biological role and mode of regulation of the production are poorly understood. In temperate eutrophic freshwaters, Microcystis and Planktothrix often co-occur, with Planktothrix being an early colonizer and Microcystis appearing subsequently. We tested if the production of a range of peptides by co-existing species could be regulated through interspecific interactions. Using a combined approach of co-cultures and analyses...
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Ano: 2017 URL: http://archimer.ifremer.fr/doc/00374/48515/48794.pdf
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Production and Isolation of Azaspiracid-1 and -2 from Azadinium spinosum Culture in Pilot Scale Photobioreactors ArchiMer
Jauffrais, Thierry; Kilcoyne, Jane; Sechet, Veronique; Herrenknecht, Christine; Truquet, Philippe; Herve, Fabienne; Berard, Jean-baptiste; Nulty, Ciara; Taylor, Sarah; Tillmann, Urban; Miles, Christopher O.; Hess, Philipp.
Azaspiracid (AZA) poisoning has been reported following consumption of contaminated shellfish, and is of human health concern. Hence, it is important to have sustainable amounts of the causative toxins available for toxicological studies and for instrument calibration in monitoring programs, without having to rely on natural toxin events. Continuous pilot scale culturing was carried out to evaluate the feasibility of AZA production using Azadinium spinosum cultures. Algae were harvested using tangential flow filtration or continuous centrifugation. AZAs were extracted using solid phase extraction (SPE) procedures, and subsequently purified. When coupling two stirred photobioreactors in series, cell concentrations reached 190,000 and 210,000 cell·mL−1 at...
Tipo: Text Palavras-chave: Solid phase extraction; Photobioreactor; Chemostat; Dinoflagellate; Micro-algae; LC-MS/MS; Tangential flow filtration; Azaspiracid; HP-20.
Ano: 2012 URL: http://archimer.ifremer.fr/doc/00088/19875/17526.pdf
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High resolution mass spectrometry for quantitative analysis and untargeted screening of algal toxins in mussels and passive samplers ArchiMer
Zendong, Suzie Zita; Mccarron, Pearse; Herrenknecht, Christine; Sibat, Manoella; Amzil, Zouher; Cole, Richard B.; Hess, Philipp.
Measurement of marine algal toxins has traditionally focussed on shellfish monitoring while, over the last decade, passive sampling has been introduced as a complementary tool for exploratory studies. Since 2011, liquid chromatography-tandem mass spectrometry (LC-MS/MS) has been adopted as the EU reference method (No.15/2011) for detection and quantitation of lipophilic toxins. Traditional LC-MS approaches have been based on low-resolution mass spectrometry (LRMS), however, advances in instrument platforms have led to a heightened interest in the use of high-resolution mass spectrometry (HRMS) for toxin detection. This work describes the use of HRMS in combination with passive sampling as a progressive approach to marine algal toxin surveys. Experiments...
Tipo: Text Palavras-chave: Monitoring; Marine toxins; Passive sampling; SPATT; Matrix effects.
Ano: 2015 URL: http://archimer.ifremer.fr/doc/00277/38848/37388.pdf
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