17 Effect of Dilution Rate on Azadinium spinosum and Azaspiracid (AZA). . .
203
Conclusion
At a rate of 0.25 day
1 , we obtained about 3 mg of AZAs in crude extracts
over 12 days (8 days of culture, 1 day of filtration and 3 days of extractions).
The optimisation of the procedure demonstrated the feasibility of producing AZAs
from A. spinosum cultured in photobioreactors in series. The AZAs obtained are
suitable amounts for purification and production of certified standards for further
toxicological study and for instrument calibration in monitoring programs.
Acknowledgment This study was carried out under the Sea Change strategy with the support
of the Marine Institute and the Marine Research Sub-Programme of the National Development
Plan 2007–2013 (ASTOX2 project PBA/AF/08/001(01)), co-financed by the European Regional
Development Fund. Further funding was obtained through Ifremer from the French Ministry of
Education, Research and Technology through Programme 187 of the National Finance Law. The
authors would like to thank all the members of the laboratory EMP/PHYC at the Atlantic Centre
of Ifremer for their help and technical advice during this study.
References
Akselman R, Negri RM (2012) Blooms of Azadinium cf. spinosum Elbr¨ achter et Tillmann
(Dinophyceae) in northern shelf waters of Argentina, Southwestern Atlantic. Harmful Algae
19:30–38. doi:10.1016/j.hal.2012.05.004
Diaz Sierra M, Furey A, Hamilton B, Lehane M, James KJ (2003) Elucidation of the fragmentation
pathways of azaspiracids, using electrospray ionisation, hydrogen/deuterium exchange, and
multiple-stage mass spectrometry. J Mass Spectrom 38:1178–1186
Fux E, Marcaillou C, Mondeguer F, Bire R, Hess P (2008) 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. Harmful Algae 7:574–583
Fux E, Bire R, Hess P (2009) Comparative accumulation and composition of lipophilic marine
biotoxins in passive samplers and in mussels (M. edulis) on the West Coast of Ireland. Harmful
Algae 8:523–537
Hernandez-Becerril DU, Escobae-Morales S, Morreno-Guti´ erez SP, Baron-Campis SA (2010)
Two new records of potentially toxic phytoplankton species from the Mexican Pacific. Poster
presented at the 14th international conference on harmful algae, Hersonissos, Nov 2010
James KJ, Sierra MD, Lehane M, Magdalena AB, Furey A (2003) Detection of five new hydroxyl
analogues of azaspiracids in shellfish using multiple tandem mass spectrometry. Toxicon
41:277–283
Jauffrais T, S´ echet V, Herrenknecht C, Tillmann U, Krock B, Amzil Z, Hess P (2010) Growth and
toxin production of Azadinium spinosum in batch and continuous culture. Poster presented at
the 14th international conference on harmful algae, Hersonissos, Nov 2010
Jauffrais T, Herrenknecht C, S´ echet V, Sibat M, Tillmann U, Krock B, Kilcoyne J, Miles CO,
McCarron P, Amzil Z, Hess P (2012) Quantitative analysis of azaspiracids in Azadinium
spinosum cultures. Anal Bioanal Chem 403:833–846
Keller MD, Selvin RC, Claus W, Guillard RRL (1987) Media for the culture of oceanic
ultraphytoplankton. J Phycol 23:633–638
Kilcoyne J, Keogh A, Clancy G, Leblanc P, Burton I, Quilliam MA, Hess P, Miles CO
(2012) Improved isolation procedure for azaspiracids from shellfish, structural elucidation of
azaspiracid-6, and stability studies. J Agric Food Chem 60:2447–2455
203
Conclusion
At a rate of 0.25 day
1 , we obtained about 3 mg of AZAs in crude extracts
over 12 days (8 days of culture, 1 day of filtration and 3 days of extractions).
The optimisation of the procedure demonstrated the feasibility of producing AZAs
from A. spinosum cultured in photobioreactors in series. The AZAs obtained are
suitable amounts for purification and production of certified standards for further
toxicological study and for instrument calibration in monitoring programs.
Acknowledgment This study was carried out under the Sea Change strategy with the support
of the Marine Institute and the Marine Research Sub-Programme of the National Development
Plan 2007–2013 (ASTOX2 project PBA/AF/08/001(01)), co-financed by the European Regional
Development Fund. Further funding was obtained through Ifremer from the French Ministry of
Education, Research and Technology through Programme 187 of the National Finance Law. The
authors would like to thank all the members of the laboratory EMP/PHYC at the Atlantic Centre
of Ifremer for their help and technical advice during this study.
References
Akselman R, Negri RM (2012) Blooms of Azadinium cf. spinosum Elbr¨ achter et Tillmann
(Dinophyceae) in northern shelf waters of Argentina, Southwestern Atlantic. Harmful Algae
19:30–38. doi:10.1016/j.hal.2012.05.004
Diaz Sierra M, Furey A, Hamilton B, Lehane M, James KJ (2003) Elucidation of the fragmentation
pathways of azaspiracids, using electrospray ionisation, hydrogen/deuterium exchange, and
multiple-stage mass spectrometry. J Mass Spectrom 38:1178–1186
Fux E, Marcaillou C, Mondeguer F, Bire R, Hess P (2008) 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. Harmful Algae 7:574–583
Fux E, Bire R, Hess P (2009) Comparative accumulation and composition of lipophilic marine
biotoxins in passive samplers and in mussels (M. edulis) on the West Coast of Ireland. Harmful
Algae 8:523–537
Hernandez-Becerril DU, Escobae-Morales S, Morreno-Guti´ erez SP, Baron-Campis SA (2010)
Two new records of potentially toxic phytoplankton species from the Mexican Pacific. Poster
presented at the 14th international conference on harmful algae, Hersonissos, Nov 2010
James KJ, Sierra MD, Lehane M, Magdalena AB, Furey A (2003) Detection of five new hydroxyl
analogues of azaspiracids in shellfish using multiple tandem mass spectrometry. Toxicon
41:277–283
Jauffrais T, S´ echet V, Herrenknecht C, Tillmann U, Krock B, Amzil Z, Hess P (2010) Growth and
toxin production of Azadinium spinosum in batch and continuous culture. Poster presented at
the 14th international conference on harmful algae, Hersonissos, Nov 2010
Jauffrais T, Herrenknecht C, S´ echet V, Sibat M, Tillmann U, Krock B, Kilcoyne J, Miles CO,
McCarron P, Amzil Z, Hess P (2012) Quantitative analysis of azaspiracids in Azadinium
spinosum cultures. Anal Bioanal Chem 403:833–846
Keller MD, Selvin RC, Claus W, Guillard RRL (1987) Media for the culture of oceanic
ultraphytoplankton. J Phycol 23:633–638
Kilcoyne J, Keogh A, Clancy G, Leblanc P, Burton I, Quilliam MA, Hess P, Miles CO
(2012) Improved isolation procedure for azaspiracids from shellfish, structural elucidation of
azaspiracid-6, and stability studies. J Agric Food Chem 60:2447–2455