Journal of Lipid Research, Vol 38, 2580-2588, Copyright © 1997 by Lipid Research, Inc.

ARTICLES

RRR-alpha-tocopherol can be substituted for by Trolox in determination of kinetic parameters of LDL oxidizability by copper

AR Bergmann, P Ramos, H Esterbauer and BM Winklhofer-Roob
Institute of Biochemistry, Karl-Franzens University of Graz, Austria.

LDL resistance to oxidation is characterized by the lag time that precedes the rapid generation of conjugated dienes in copper-induced oxidation. Lag time (y) is described by the equation y = kx + a, where k is the efficacy constant of alpha-tocopherol (alpha-TOH), x the alpha- TOH content of LDL, and a the alpha-TOH-independent variable, all of which show considerable between-subject variability. To answer the question of whether loading of LDL with Trolox can substitute for the more timeconsuming loading with RRR-alpha-TOH in large scale studies focusing on determination of k and a values, LDL from 15 healthy subjects was loaded in vitro with 0-50 mol Trolox/mol LDL or 0-250 mM RRR-alpha-TOH. In addition, five of these subjects were supplemented in vivo with 800-1200 IU/d RRR-alpha-TOH for 17 days and k and a were determined. The ratios k(Trolox):k(alpha-TOH) and alpha(Trolox):a(alpha- TOH) were calculated. The k(Trolox) was only slightly higher than k(alpha-TOH), in contrast to alpha(Trolox), which was almost twice as high as a(alpha-TOH), indicating that, in the case of Trolox, a includes the contribution of alpha-TOH to the lag time. In the case of alpha-TOH, a includes only the additional antioxidants that occur naturally in LDL. The ratios k(Trolox):k(alpha-TOH) and alpha(Trolox):a(alpha-TOH) showed little variability, both between subjects and between in vitro and in vivo experiments. A close correlation was found between k(alpha-TOH) and a(alpha0TOH), calculated using this ratio, and k(alpha-TOH) and a(alpha-TOH), obtained in our experiments. We conclude that from this ratio, k(alpha-TOH) and a(alpha- TOH) can be reliably calculated when k(Trolox) and a(Trolox) are determined experimentally, thus allowing us to obtain k and a values in a more convenient way.
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