Abstract
The spirolides are a family of marine biotoxins derived from the dinoflagellate Alexandrium ostenfeldii, recently isolated from contaminated shellfish and characterized. A crude phytoplankton extract has been extensively studied for mass spectrometric determination and characterization of several known spirolides and previously unreported compounds. The complex sample was initially analyzed by full-scan mass spectrometry in an ion-trap instrument, enabling identification of several components. Subsequent analysis by selected-ion monitoring in a triple-quadrupole instrument resulted in the confirmation of the identities of the compounds detected in the ion trap. Purification of the crude extract was performed using an automated mass-based fractionation system, yielding several fractions with different relative contributions of the spirolide components. Collision-induced dissociation (CID) in the triple-quadrupole instrument produced significant fragment ions for all identified species. Selective enrichment of some minor compounds in certain fractions enabled excellent CID spectra to be generated; this had previously been impossible, because of interferences from the major toxins present. Fourier-transform ion cyclotron resonance (FTICR) mass spectrometry was then performed for accurate determination of the masses of MH+ ions of all the species present in the sample. Additionally, infrared multiphoton dissociation in the FTICR instrument generated elemental formulae for product ions, including those formed in the previous collisional activation experiments. Collection of these results and the fragmentation scheme proposed for the main component of the extract, 13-desmethyl spirolide C, from part I of this study, enabled elucidation of the structures of some uncharacterized spirolides and some biogenetically related compounds present at previously unreported masses.
Similar content being viewed by others
References
Cembella AD, Bauder AG, Lewis NI, Quilliam MA (2001) J Plankton Res 23:1413–1419
Gribble KE, Keafer BA, Kulis DM, Cembella AD, Anderson DM (2003) In: Proc 10th Int Conf on Harmful Algae, October 21–25, 2002, St Pete Beach, Florida, USA, p 116
MacKinnon SL, Cembella AD, Quilliam MA, LeBlanc P, Lewis NI, Hardstaff WR, Burton IW, Walter JA (2003) In: Proc 10th Int Conf on Harmful Algae, October 21–25, 2002, St Pete Beach, Florida, USA, p 180
Rühl A, Hummert C, Reinhardt K, Gerdts G, Luckas B (2001) In: Proc Int Council for the Exploration of the Sea (ICES), Oslo, Norway 2001/S:09
Hu T, Curtis JM, Oshima Y, Quilliam MA, Walter JA, Watson-Wright WM, Wright JLC (1995) J Chem Soc Chem Commun 2159–2161
Cembella AD, Quilliam MA, Lewis NI, Bauder AG, Wright JLC (1998) Harmful Algae 481–484
Richard D, Arsenault E, Cembella A, Quilliam M (2000) Harmful Algal Blooms 383–386
Cembella AD, Lewis NI, Quilliam MA (1999) Nat Toxins 7:197–206
Cembella AD, Lewis NI, Quilliam MA (2000) Phycologia 39:67–74
Cembella AD, Bauder AG, Lewis NI, Quilliam MA (2000) In: Hallegraff GM, Blackburn SI, Bolch CJ, Lewis RJ (eds) Harmful algal blooms, Intergovernmental Oceanographic Commission of UNESCO, 2001, 173–176
Hu T, Burton IW, Cembella AD, Curtis JM, Quilliam MA, Walter JA, Wright JLC (2001) J Nat Prod 64:308–312
Hu T, Curtis JM, Walter JA, Wright JLC (1996) Tetrahedron Lett 37:7671–7674
Falk M, Burton IW, Hu T, Walter JA, Wright JLC (2001) Tetrahedron 57:8659–8665
Wiltshire H (2000) In: Venn RF (ed) Principles and practice of bioanalysis. Taylor and Francis, New York, pp 325–326
Quilliam MA (2003) J Chromatogr A 1000:527–548
Hummert C, Rühl A, Reinhardt K, Gerdts G, Luckas B (2002) Chromatographia 55:673–680
Sleno L, Windust AJ, Volmer DA (2003) Anal Bioanal Chem, in print
Cembella AD, Lewis NI, Quilliam MA (1999) Nat Toxins 7:197–206
Senko MW, Canterbury JD, Guan S, Marshall AG (1996) Rapid Commun Mass Spectrom 10:1839–1844
Yates NA, Booth MM, Stephenson JL Jr, Yost RA (1995) In: March RE, Todd JFJ (eds) Practical aspects of ion trap mass spectrometry, vol III. Chemical, environmental and biomedical applications. CRC Press, Boca Raton, FL, USA, p 147
Bier ME, Schwartz JC (1997) In: Cole RB (ed) Electrospray ionization mass spectrometry: fundamentals, instrumentation and applications. Wiley, New York, pp 253–254
Acknowledgements
The authors wish to thank Dr Anthony Windust (NRC/IMB, Halifax, NS, Canada) for the culture of the phytoplankton and help with the partial purification of the extract, and Dr Kevin Bateman, Carmai Seto and Tammy LeRiche at the Merck Frosst Centre for Therapeutic Research (Kirkland, QC, Canada) for invaluable help in the mass-triggered fractionation experiments. LS would also like to acknowledge financial assistance from le Fonds Quebecois de la recherche sur la nature et les technologies (FCAR), the National Research Council’s Graduate Student Scholarship Supplement Program (GSSSP) and Dr Alan Cembella at the National Research Council’s Institute for Marine Biosciences. This work was supported in part by the NSF National High-Field FTICR MS Facility, CHE-99-09502
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Sleno, L., Chalmers, M.J. & Volmer, D.A. Structural study of spirolide marine toxins by mass spectrometry. Anal Bioanal Chem 378, 977–986 (2004). https://doi.org/10.1007/s00216-003-2296-0
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00216-003-2296-0
Keywords
- Ion-trap mass spectrometry
- Triple-quadrupole mass spectrometry
- Fourier-transform ion cyclotron resonance mass spectrometry
- Full-scan mass spectrometry
- Selected-ion monitoring
- Collision-induced dissociation
- Infrared multiphoton dissociation
- Automated mass-based fractionation
- Phytoplankton
- Spirolide marine toxins
- Alexandrium ostenfeldii