Summary
The RP-HPLC analysis of chlorophylls, their degradation products (chloropigments) and carotenoids is very sensitive to the nature of the injection solvent. The effect of sample-solvent interaction can result in the production of distorted, or even false, peaks that could be erroneously interpreted as “pigment like” or as poor chromatographic resolution. The previously suggested theoretical explantion, based on differences in solvent characteristics as expressed by the Polarity Index (P′) or even by the more precise solvent strength parameter for reversed-phase systems (S) we use, was unsatisfactory. The problem seems more complex, with other parameters such as injection volume and solute concentration or solvent selectivity also playing a role. As a practical consequence, however, suggesions are made for optimum injection conditions. Also, the presence of ion-pairing reagents in the injection solvent is demonstrated to be unnecessary.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
W. A. Svec, in “The porphyrins”, D. Dolphin, ed., Academic Press, London, 1978; vol. V, p. 341.
T. W. Goodwin, G. Britton, in “Plant Pigments”. J. W. Goodwin, ed. Academic Press, London, 1988; p. 62.
S. Roy, J. Chromatogr.391, 19 (1987).
S. W. Wright, J. D. Shearer, Chromatogr.294, 281 (1984).
M. Zapata, Ana M. Ayala, J. M. Franco, J. L. Garrido, Chromatographia23, 26 (1987).
W. W. C. Gieskes, G. W. Kraay, A. Nontij, D. Setiapermana, A. Sutomo, Neth. J. Sea Res.22, 123 (1988).
L. R. Snyder, J. J. Kirkland, in “Introduction to Modern Liquid Chromatography”, 2nd ed., New York, 1979; p. 246.
M. Tsimidou, R. Macrae, J. Chromatogr. Sci.23, 155 (1985).
F. Khachik, G. R. Beecher, J. T. Vanderslice, G. Furrow, Anal. Chem.60, 807 (1988).
L. R. Snyder, J. W. Dolan, J. R. Gant, J. Chromatogr.165, 3 (1979).
R. F. C. Mantoura, C. A. Llewellyn, Anal. Chim. Acta151, 297 (1983).
C. K. Lim, T. J. Peters, J. Chromatogr.316, 397 (1984).
C. Horvath, W. Melander, I. Molnar, P. Molnar, Anal. Chem.49, 2295 (1977).
L. Rohrschneider, Anal. Chem.45, 1241 (1973).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Zapata, M., Garrido, J.L. Influence of injection conditions in reversed-phase high-performance liquid chromatography of chlorophylls and carotenoids. Chromatographia 31, 589–594 (1991). https://doi.org/10.1007/BF02279480
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02279480