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A fluorescence method for the determination of plasma susceptibility to lipid peroxidation
G Cervato1, P Viani, R Cazzola
1Department of Medical Chemistry and Biochemistry, L.I.T.A. Vialba, University of Milan, Milano, Italy. cestaro@imiucca.csi.unimi.it
Objective:
We propose a fluorescence kinetics method for monitoring plasma susceptibility to peroxidation.
Design And Method:
Plasmatic peroxidation was induced by CuSO4 (500 microM), and fluorescence was measured every 30 min. Kinetics were represented by a sigmoidal curve from which it was possible to calculate the latency time (lag-time) and the propagation velocity (slope) of plasma peroxidation.
Results:
The lag-time monitored by the fluorescence kinetics method corresponded to the formation of thiobarbituric acid reactive substances, and to progressive depletion of polyunsaturated fatty acids and alpha-tocopherol. The mechanism of reaction appeared to be dependent upon plasmatic hydroperoxides, and independent of oxygen radicals. Plasma storage is possible for at least two months at -80 degrees C, and reproducibility of the method is very good.
Conclusions:
Fluorescence kinetics provide a highly comprehensive picture of plasma susceptibility to peroxidation in comparison with the conventional measurements of anti- and pro-oxidant ratios.