Abstract
Lipid peroxides in serum were determined by use of a high performance liquid chromatograph (HPLC) with fluorescence detection and 1, 3-diphenyl-2-thiobarbituric acid (DPTBA) instead of thiobarbituric acid (TBA). Optimum analytical conditions for lipid peroxides were determined by using serum and tetramethoxypropane (IMP) as the standard aqueous solution of malondialdehyde (MDA). The optimum pH for the reaction of TMP with DPTBA was found to be 2.5. The lipid peroxides and bilirubin, which reacts with DPTBA and interferes with the measurement of lipid peroxides, were separated by use of HPLC. A linear relation passing through the origin point was obtained between the fluorescence intensity and the concentration of MDA standard solution over the range of 0.06-0.24nmol/ml. The relative standard deviation was 2.21% (n=5). The detection limit of the lipid peroxides in serum was 12pmol/ml as S/N=10. The recovery of 0.6nmol MDA added to 50μl of serum was 97.8%.
The sensitivity of HPLC-fluorescence detection method was 17 times greater than that of the HPLC-visible detection method and 192 times higher than that of the TBA method.
