Abstract
The suitability of N,N-dimethylformamide (DMF) as an extractant for the standard reverse-phase HPLC method was examined using algal cultures. Good pigment separations and recovery were achieved with 20% (volume %) addition of an ion-pairing solution in an injection. While slight amounts of degradation products of chlorophylla, i.e., chlorophyllidea, allomeric and epimeric forms, were produced, adequate attention to filtration and extraction prevents the formation of degradation products, confining them to an acceptable level. Because of its strong extractability, which expedites the extraction process, DMF is an efficient solvent for HPLC analysis of phytoplankton pigments.
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References
Barrett, J. and S. W. Jeffrey (1971): A note on the occurrence of chlorophyllase in marine algae.J. Exp. Mar. Biol. Ecol.,7, 252–262.
Gieskes, W. W. C. (1991): Algal pigment fingerprints: Clue to taxon-specific abundance, productivity and degradation of phytoplankton in seas and oceans. p. 61–99. InParticle Analysis in Oceanography, ed. by S. Demers, Springer-Verlag, Berlin.
Guillard, R. R. L. and J. H. Ryther (1962): Studies of marine planktonic diatoms. I.Cyclotella nana Hustedt andDetonula confervacea (Cleve) gran.Can. J Microbiol.,8, 229–239.
Jeffrey, S. W. and G. M. Hallegraeff (1987): Chlorophyllase distribution in ten classes of phytoplankton: a problem for chlorophyll analysis.Mar. Ecol. Prog. Ser.,35, 293–304.
Li, W. K. W. (1986). Experimental approaches to field measurements: methods and interpretation. p. 251–286 InPhotosynthetic Picoplankton, ed. by T. Platt and W. K. W. Li,Can. Bull. Fish. Aquat. Sci.,214, Dept. Fish and Ocean, Ottawa.
Mantoura, R. F. C. and C. A. Llewellyn (1983): The rapid determination of algal chlorophyll and carotenoid pigments and their breakdown products in natural waters by reversephase high-performance liquid chromatography.Anal. Chim, Acta,151, 297–314.
Mantoura, R. F. C., S. W. Wright, S. W. Jeffrey, R. G. Barlow and D. E. Cummings (1997): Filtration and storage of pigments from microalgae. p. 283–305. InPhytoplankton Pigments in Oceanography, ed. by S. W. Jeffrey, R. F. C. Mantoura and S. W. Wright, Monographs on oceanographic methodology UNESCO, Paris.
Moran, R. and D. Porath (1980): Chlorophyll determination in intact tissues using N,N-dimethylformamide.Plant Physiol.,65, 478–479.
Speziale, B. J., S. P. Schreiner, P. A. Giammatteo and J. E. Schindler (1984): Comparison of N,N-dimethylformamide, dimenthylsulfoxide, and acetone for extraction of phytoplankton chlorophyll.Can. J Fish. Aquat. Sci.,41, 1519–1522.
Suzuki, R. and Y. Fujita (1986): Chlorophyll decomposition inSkeletonema costatum: a problem in chlorophyll determination of water samples.Mar. Ecol. Progr. Ser.,28, 81–85.
Suzuki, R. and T. Ishimaru (1990): An Improved method for the determination of phytoplankton chlorophyll using N,N-dimethylformamide.J. Oceanogr. Soc. Japan,46, 190–194.
Suzuki, R., M. Takahashi, K. Furuya and T. Ishimaru (1993): Simplified technique for the rapid determination of phytoplankton pigments by reverse-phase high-performance liquid chromatography.J. Oceanogr.,49, 571–580.
Watanabe, T., H. Mazaki and M. Nakazato (1987): Chlorophylla/a′ epimerization in organic solvents.Biochim. Biophys. Acta,892, 197–206.
Wright, S. W., S. W. Jeffrey, r. F. C. Mantoura, C. A. Llewellyn, T. Bjornland, D. Repeta and N. Welschmeyer (1991): Improved HPLC method for the analysis of chlorophylls and carotenoids from marine phytoplankton.Mar. Ecol. Prog. Ser.,77, 183–196.
Wright, S. W., S. W. Jeffrey and R. F. C. Mantoura (1997): Evaluation of methods and solvents for pigment extraction. p. 261–282. InPhytoplankton Pigments in Oceanography, ed. by S. W. Jeffrey, R. F. C. Mantoura and S. W. Wright, Monographs on oceanographic methodology, UNESCO, Paris.
Zapata, M., A. M. Ayala, J. M. Franco and J. L. Garrido (1987): Separation of chlorophylls and their degradation products in marine phytoplankton by reversed-phase high-performance liquid chromatography.Chromatogr.,23, 26–30.
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Furuya, K., Hayashi, M. & Yabushita, Y. HPLC determination of phytoplankton pigments using N,N-dimethylformamide. J Oceanogr 54, 199–203 (1998). https://doi.org/10.1007/BF02751695
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DOI: https://doi.org/10.1007/BF02751695