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The Use of HPLC for the Characterization of Phytoplankton Pigments

  • José L. Garrido
  • Suzanne Roy
Part of the Methods in Molecular Biology book series (MIMB, volume 1308)

Abstract

HPLC is still the technique of choice for the analysis and characterization of phytoplankton pigments. In this chapter we describe procedures for sample preparation and pigment extraction, and the use of octyl silica columns and pyridine-containing mobile phases to separate chlorophylls and carotenoids. The identification of pigments on the basis of their retention times and visible spectra, the preparation of pigment standards, and the quantitative analysis by either external or internal standard procedures are also described.

Key words

Carotenoids Chlorophylls HPLC Octyl silica Photodiode array detector Phytoplankton 

References

  1. 1.
    Wright SW, Jeffrey SW (2006) Pigment markers for phytoplankton production. In: Volkman JK (ed) Marine organic matter, vol 2, Handbook of environmental chemistry. Springer, Heidelberg, pp 71–104Google Scholar
  2. 2.
    Wright SW (2005) Analysis of phytoplankton populations using pigment markers. http://www.antarctica.gov.au/__data/assets/pdf_file/0007/21877/ml_38651523287037_analysis20of20phytoplankton20pigments.pdf. Accessed 1 Nov 2013
  3. 3.
    Roy S, Llewellyn CA, Egeland E et al (eds) (2011) Phytoplankton pigments: characterization, chemotaxonomy and applications in oceanography. Cambridge University Press, Cambridge, UKGoogle Scholar
  4. 4.
    Jeffrey SW, Mantoura RFC, Wright SW (eds) (1997) Phytoplankton pigments in oceanography: guidelines to modern methods. UNESCO Publishing, ParisGoogle Scholar
  5. 5.
    Roy S, Garrido JL (2013) Pigments Liquid chromatography. In: Reedijk J (ed) Reference module in chemistry, molecular sciences and chemical engineering. Elsevier, Waltham, MA. doi: 10.1016/B978-0-12-409547-2.04878-2 Google Scholar
  6. 6.
    Zapata M, Rodríguez F, Garrido JL (2000) Separation of chlorophylls and carotenoids from marine phytoplankton: a new HPLC method using a reversed-phase C8 column and pyridine-containing mobile phases. Mar Ecol Prog Ser 195:29–45CrossRefGoogle Scholar
  7. 7.
    Van Heukelem L, Thomas CS (2001) Computer-assisted high-performance liquid chromatography method development with applications to the isolation and analysis of phytoplankton pigments. J Chromatogr A 910:31–49PubMedCrossRefGoogle Scholar
  8. 8.
    Garrido JL, Airs R, Rodríguez F et al (2011) New HPLC separation techniques. In: Roy S, Egeland ES, Johnsen G et al (eds) Phytoplankton pigments: characterization, chemotaxonomy and applications in oceanography. Cambridge University Press, Cambridge, UK, pp 165–194CrossRefGoogle Scholar
  9. 9.
    Roy S, Wright SW, Jeffrey SW (2011) Phytoplankton cultures for standard pigments and their suppliers. In: Roy S, Egeland ES, Johnsen G et al (eds) Phytoplankton pigments: characterization, chemotaxonomy and applications in oceanography. Cambridge University Press, Cambridge, UK, pp 653–657CrossRefGoogle Scholar
  10. 10.
    Zapata M, Garrido JL (1991) Influence of injection conditions in reversed-phase high-performance liquid chromatography of chlorophylls and carotenoids. Chromatographia 31:589–594CrossRefGoogle Scholar
  11. 11.
    Egeland ES (2011) Data sheets aiding identification of phytoplankton carotenoids and chlorophylls. In: Roy S, Egeland ES, Johnsen G et al (eds) Phytoplankton pigments: characterization, chemotaxonomy and applications in oceanography. Cambridge University Press, Cambridge, pp 665–822Google Scholar
  12. 12.
    Zapata M, Rodríguez F, Garrido JL et al (2004) Photosynthetic pigments in 37 species (65 strains) of Haptophyta: implications for oceanography and chemotaxonomy. Mar Ecol Prog Ser 270:83–102CrossRefGoogle Scholar
  13. 13.
    Zapata M, Fraga S, Rodríguez F et al (2012) Pigment-based chloroplast types in dinoflagellates. Mar Ecol Prog Ser 465:33–52CrossRefGoogle Scholar
  14. 14.
    Mantoura RFC, Repeta D (1997) Calibration methods for HPLC. In: Jeffrey SW, Mantoura RFC, Wright SW (eds) Phytoplankton pigments in oceanography: guidelines to modern methods. UNESCO Publishing, Paris, pp 407–428Google Scholar
  15. 15.
    Van Heukelem L, Hooker SB (2011) The importance of the quality assurance plan for method validation and minimizing uncertainties in the HPLC analysis of phytoplankton pigments. In: Roy S, Egeland ES, Johnsen G et al (eds) Phytoplankton pigments: characterization, chemotaxonomy and applications in oceanography. Cambridge University Press, Cambridge, pp 195–242CrossRefGoogle Scholar
  16. 16.
    Hooker SB, Van Heukelem L (2011) A symbology and vocabulary for an HPLC lexicon. In: Roy S, Egeland ES, Johnsen G et al (eds) Phytoplankton pigments: characterization, chemotaxonomy and applications in oceanography. Cambridge University Press, Cambridge, pp 243–256Google Scholar
  17. 17.
    Pinckney JL, Millie DF, Van Heukelem L (2011) Update on filtration, storage and extraction solvents. In: Roy S, Egeland ES, Johnsen G et al (eds) Phytoplankton pigments: characterization, chemotaxonomy and applications in oceanography. Cambridge University Press, Cambridge, pp 627–635CrossRefGoogle Scholar
  18. 18.
    Alou-Font E, Mundy CJ, Roy S et al (2013) Snow cover affects ice algal pigment composition in the coastal Arctic Ocean during spring. Mar Ecol Prog Ser 474:89–104CrossRefGoogle Scholar
  19. 19.
    Rousseaux G, Martin M, De Wit A (2011) Viscous fingering in packed chromatographic columns: non-linear dynamics. J Chromatogr A 1218:8353–8361PubMedCrossRefGoogle Scholar
  20. 20.
    Latasa M, van Lenning K, Garrido JL et al (2001) Losses of chlorophylls and carotenoids in aqueous acetone and MeOH extracts prepared for RPHPLC analysis of pigments. Chromatographia 53:385–391CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, New York 2015

Authors and Affiliations

  1. 1.Instituto de Investigaciones Marinas (CSIC)VigoSpain
  2. 2.ISMERUniversité du Québec à RimouskiRimouskiCanada

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