Ferrothiocyanate법에 의한 혈청 Lipid Hydroperoxide정량

Quantitative Determination of Lipid Hydroperoxide in Human Blood Serum by Ferrothiocyanate Method

In order to develope the method of quantitative determination of lipid hydroperoxide in human blood serum, we tried the ferrothiocyanate method to total lipids extracted by Bligh-Dyer method and obtained the results as follows. 1. The maximum absorbance showed at the concentration of Mohr s solution, 0.127M at pH 1.70 and ammonium thiocyanate solution, 3.95M in the ferrothiocyanate method. 2. When hydrogen peroxide, cumene hydroperoxide, and oxidized linoleic acid were added to serum, and extracted them by Bligh-Dyer method to examine the extraction efficiency, we confirmed that cumene hydroperoxide and oxidized linoleic acid were extracted in CHCI₃ phase, and hydrogen peroxide in MeOH-H₂O phase, respectively. 3. The concentration of lipid hydroperoxide of total lipids extracted from normal adult serum was 2.0×10⁻⁵M, and increased proportionally the concentration of lipid hydroperoxide by increasing the amount of serum. 4. When we compared the total lipids extracted by Bligh-Dyer method and total lipids extracted after lipoprotein is precipitated by Yagi method in human blood serum, the concentration of lipid hydroperoxide was showed nearly the same value. From our results, we concluded that the concentration of lipid hydroperoxide in human blood serum could be determined quantitatively by ferrothiocyanate method.