Abstract
Kinetics of formation of fluorescent condensation products from hexanal andl-lysine (or itsN-acetylated forms) including mass-transfer has been studied in a two-phase system consisting of lysine (or lysine derivative) in a aqueous phosphate buffer and a 1-octanol solution of hexanal as model for formation of fluorophores between protein and carbonyl compounds in peroxidizing biological systems. The initial rate of formation of fluorescent products in the aqueous phase was found to be proportional to the concentration of hexanal and lysine and to increase in both phases with increasing pH in the aqueous phase, in contrast to a higher-order dependence on hexanal in the octanol phase. At pH=6.8, the temperature dependence of the appearance of fluorescent products corresponds to apparent energies of activation of 63 kJ·mol−1 and 87 kj·mol−1 in the aqueous phase and the octanol phase, respectively. Fluorescent condensation products appeared faster in the octanol phase. However, by a kinetic analysis, the fluorescent products were shown to be formed in the aqueous phase, corresponding to the lower energy of activation and to the simple second-order kinetics, and subsequently distributed between the aqueous phase and the octanol phase.l-Lysine reacted faster thanN α-acetyl-l-lysine which reacted faster thanN ε-acetyl-l-lysine. Using fluorescence quantum yields, determined to be 1.4·10−2 in octanol and 8·10−3 in water at pH 6.8, an apparent partition coefficient of 17 (octanol/water) was determined for the condensation product ofl-lysine. The steady-state fluorescence in the octanol phase was attributed to two components with fluorescence life-time at 25°C of 0.7±0.05 ns and 5.1±0.2 ns, assigned to hexanal and the condensation product, respectively. The emission spectra, were resolved in the two components using phase-sensitive detection, and the condensation product had emission maximum at 405 nm.
Similar content being viewed by others
Abbreviations
- Dimethyl-POPOP:
-
1,4-bis(4-methyl-5-phenyl-oxazolyl)benzene
References
Montgomery, M.W., and Day, E.A. (1965) Aldehyde-Amine Condensation Reaction: A Possible Fate of Carbonyls in Foods,J. Food Sci. 30, 828–832.
Kikugawa, K. (1986) Fluorescent Products Derived from the Reaction of Primary Amines and Components in Peroxidized Lipids,Adv. Free Radical Biol. Med. 2, 389–417.
Chio, K.S., Reiss, U., Fletcher, B., and Tappel, A.L. (1969) Peroxidation of Subcellular Organelles: Formation of Lipofuscin-like Fluorescent Pigments,Science 166, 1535–1536.
Dillard, C., and Tappel, A.L. (1971) Fluorescent Products of Lipid Peroxidation of Mitochondria and Microsomes,Lipids 6, 715–721.
Gray, J.I. (1978) Measurement of Lipid Oxidation: A Review,J. Am. Oil Chem. Soc. 55, 539–546.
Stapelfeldt, H., Bjørn, H., Skovgaard, I.M., Skibsted, L.H., and Bertelsen, G. (1992) Warmed-Over Flavour in Cooked Sliced Beef. Chemical Analysis in Relation to Sensory Evaluation,Z. Lebensm. Unters.-Forsch. 195, 203–208.
Guthrie, F.E., and Hodgson, E. (1987) Absorption and Distribution of Toxicants, inA Textbook of Modern Toxicology (Hodgson, E., and Levi, P.E. eds.) pp. 23–50, Elsevier, New York.
Esterbauer, H. (1982) Aldehydic Products of Lipid Peroxidation, inFree Radicals, Lipid Peroxidation and Cancer (McBrien, D.C.H., and Slater, T.F., eds.) pp. 101–128, Academic Press, London.
Elleder, M. (1981) Chemical Characterization of Age Pigments, inAge Pigments (Sohal, R.S., ed.) pp. 204–241, Elsevier, Amsterdam.
Chio, K.S., and Tappel, A.L. (1969) Inactivation of Ribonuclease and Other Enzymes, by Peroxidizing Lipids and by Malonaldehyde,Biochemistry 8, 2827–2832.
Kikugawa, K., Maruyama, T., Machida, Y., and Kurechi, T. (1981) Studies on Peroxidized Lipids. II. Fluorescent Products Relevant to Aging Pigmetns Derived from Malondialdehyde and Methylamine,Chem. Pharm. Bull. 29, 1423–1432.
Kikugawa, K., Machida, Y., Kida, M., and Kurechi, T. (1981) Studies on Peroxidized Lipids. III. Fluorescent Pigments Derived from the Reaction of Malonaldehyde and Amino Acids,Chem. Pharm. Bull 29, 3003–3011.
Eldred, G.E., and Katz, M.L. (1989) The Autofluorescent Products of Lipid Peroxidation May Not Be Lipofuscin-Like,Free Radical Biol. Med. 7, 157–163.
Minamide, Y., Horie, T., Itoh, F., and Awazu, S. (1990) Fluorescence Lifetimes of Fluorescent Substances Formed in Peroxidized Microsomes of Rat Liver,Analyst. Lett. 23, 57–65.
Minamide, Y., Horie, T., and Awazu, S. (1992) Fluorospectroscopic Analysis of the Fluorescent Substances in Peroxidized Microsomes of Rat Liver,Lipids 27, 354–359.
Parker, C.A. (1968)Photoluminescence of Solutions. Elsevier, Amsterdam.
Demas, J.N., and Crosby, G.A. (1971) The Measurement of Photoluminescence Quantum Yields: A Review,Journal of Physical Chemistry.75, 991–1024.
Melhuish W.H. (1961) Quantum Efficiencies of Fluorescence of Organic Substances: Effects of Solvent and Concentration of the Fluorescent Solute,J. Phys. Chem. 65, 229–235.
Riddick, J.A., Bunger, W.B., and Sakano, T.K. (1986)Organic Solvents (Techniques of Chemistry, Vol. II), Wiley, New York.
Lakowicz, J.R. (1983),Principles of Fluorescence Spectroscopy, Plenum, New York.
Lakowicz, J.R., Cherek, H., and Balter, A. (1981) Correction of Timing Errors in Photomultiplier Tubes Used in Phase-Modulation Fluorometry,J. Biochem. Biophys. Methods 5, 131–146.
Lakowicz, J.R., Laczko, G., Cherek, H., Gratton, E., and Limkeman, M. (1984) Analysis of Fluorescence Decay Kinetics from Varible-Frequency Phase Shift and Modulation Data,Biophys. J. 46, 463–477.
Lakowicz, J., and Cherek, H. (1981) Phase-Sensitive Fluorescence Spectroscopy: A New Method to Resolve Fluorescence Lifetimes or Emission Spectraof Components in a Mixture of Fluorophores,J. Biochem. Biophys. Meth. 5, 19–35.
Leahy, D.E. (1986) Intrinsic Molecular Volume as a Measure of the Cavity Term in Linear Solvation Energy Relationships: Octanol-Water Partition Coefficients and Aqueous Solubilities,J. Pharm. Sci. 75, 629–636.
Kikugawa, K., Iwata, A., and Beppu, M. (1988) Formation of Cross-Links and Fluorescence in Polylysine, Soluble Proteins and Membrane Proteins by Reaction with 1-Butanal,Chem. Pharm. Bull. 36, 685–692.
Espenson, J.H. (1981)Chemical Kinetics and Reaction Mechanisms, McGraw-Hill, New York.
Davis, P.L. (1968) Determination of Solubilities of C5−C9 Aldehydes in Water by Gas Chromatography,J. Gas Chromatogr. 6, 518–519.
Pliška, V., Schmidt, M., and Fauchère, J.-L. (1981) Partition Co-efficients of Amino Acids and Hydrophobic Parameters π of Their Side-Chains as Measured by Thin-Layer Chromatography.J. Chromatogr. 216, 79–92.
Martell, A.E., and Smith, R.M. (1974)Critical Stability Constants, Vol. 1: Amino Acids, Plenum, New York.
Gilbert, A., and Baggott, J. (1991)Essentials of Molecular Photochemistry, Blackwell, Oxford.
Kossanyi, J., Daccord, G., Sabbah, S., Furth, B., Chaquin, P., André, J.C. and Bouchy, M. (1980) Photochemistry in Solution. Self-Quenching and Quenching by Dienes of Aldehyde Singlet States,Nouv. J. Chim. 4, 337–344.
Kikugawa, K., and Sawamura, A. (1987) Formation of Fluorescent Substances in Reaction of Aliphatic Aldehydes and Methylamine.J. Am. Oil Chem. Soc. 64, 1156–1162.
Kikugawa, K., Takayanagi, K., and Watanabe, S. (1985) Polylysines Modified with Malonaldehyde, Hydroperoxylinoleic Acid and Monofunctional Aldehydes,Chem. Pharm. Bull. 33, 5437–5444.
Beppu, M., Murakami, K., and Kikugawa K. (1986) Fluorescent and Cross-Linked Proteins of Human Erythrocyte Ghosts Formed by Reaction with Hydroperoxylinoleic Acid, Malonaldehyde and Manofunctional Aldehydes,Chem. Pharm. Bull. 34, 781–788.
Yamaki, S., Kato, T., and Kikugawa, K. (1992) Characteristics of Fluorescence Formed by the Reaction of Proteins with Unsaturated Aldehydes, Possible Degradation Products of Lipid Radicals,Chem. Pharm. Bull. 40, 2138–2142.
Suyama, K., and Adachi, S. (1979) Reaction of Alkanals and Amino Acids or Primary Amines. Synthesis of 1,2,3,5- and 1,3,4,5-Substituted Quaternary Pyridinium Salts,J. Org. Chem. 44, 1417–1420.
Kikugawa, K., Kato, T., and Iwata, A. (1989) A Tetrameric Dialdehyde Formed in the Reaction of Butyraldehyde and Benzylamine: A Possible Intermediary, Component for Protein Cross-Linking Induced by Lipid Oxidation,Lipids 24, 962–968.
Fukuzawa, K., Kishikawa, K., Tokumura, A., Tsukatani, H., and Shibuya, M. (1985) Fluorescent Pigments by Covalent Binding of Lipid Peroxidation By-Products to Protein and Amino Acids,Lipids 20, 854–861.
Esterbauer, H., Zollner, H., and Scholz, N. (1975) Reaction of Glutathione with Conjugated Carbonyls,Z. Naturforsch. 30C, 466–473.
Author information
Authors and Affiliations
About this article
Cite this article
Stapelfeldt, H., Skibsted, L.H. Kinetics of formation of fluorescent products from hexanal andl-lysine in a two-phase system. Lipids 31, 1125–1132 (1996). https://doi.org/10.1007/BF02524286
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02524286