Effect of temperature, cholesterol content, and antioxidant structure on the mobility of vitamin E constituents in biomembrane models studied by laterally diffusion-controlled fluorescence quenching.

Kinetic parameters relevant for the antioxidant activity of the vitamin E constituents (alpha, beta, gamma, and delta homologues of tocopherols and tocotrienols) and of an amphiphilic vitamin C derivative, l-ascorbyl 6-palmitate, were determined. Fluorescence quenching experiments of 2,3-diazabicyclo[2.2.2]oct-2-ene in homogeneous acetonitrile-water mixtures afforded reactivity trends in terms of intermolecular quenching rate constants, while the quenching of Fluorazophore-L in liposomes provided the lateral diffusion coefficients relevant for understanding their biological activity in membranes. The reactivity in homogeneous solution was not influenced by the nature of the isoprenoid tail (tocopherol versus tocotrienol), but was dependent on the methylation pattern. The resulting order (alpha > beta = gamma > delta) was found to be in line with their reactivities toward peroxyl radicals as well as the phenolic O-H bond dissociation energies. The mutual lateral diffusion coefficient in POPC liposomes was the same, within error, for different tocopherols and tocotrienols (D(L) = (1.6 +/- 0.2) x 10(-7) cm(2) s(-1)). l-Ascorbyl 6-palmitate exhibited a reactivity similar to that of delta-tocopherol in homogeneous solution, but displayed a 1 order of magnitude lower fluorescence quenching efficiency in liposomes than the vitamin E constituents. Temperature effects on the laterally diffusion-controlled fluorescence quenching were large, with activation energies of 44 +/- 6 kJ mol(-1). The addition of cholesterol (0-30%) to POPC liposomes resulted only in slightly reduced diffusion coefficients. The combined results demonstrate that Fluorazophore-L can provide important physicochemical parameters for the understanding of antioxidant activity in biological environments.