Kayden H J, Chow C K, Bjornson L K
J Lipid Res. 1973 Sep;14(5):533-40.
A relatively rapid procedure is described for the spectrophotometric determination of total tocopherol in red blood cells (RBC) based on a modification of the original Emmerie-Engel reaction. The critical feature in this method is the presence of a large amount of an added antioxidant, pyrogallol or ascorbic acid, during the saponification and extraction stages and the use of thin-layer chromatography for tocopherol purification. The total tocopherol levels of plasma and erythrocytes were determined for a number of human subjects, for patients with abetalipoproteinemia, and for rats. It was found that these levels had a wide range in normal human subjects but that the ratio of RBC to plasma tocopherol was relatively constant and equal to 0.18, uncorrected, and 0.21 when both RBC and plasma values were corrected to 100% recovery. The RBC-to-plasma ratio for rats was 0.39. The accuracy of this ratio determined by the spectrophotometric procedure was verified by measuring the distribution of [(14)C]tocopherol in RBC and plasma when radioactive vitamin E was introduced into the blood by both in vitro and in vivo techniques. The addition of radioactive tocopherol to RBC or plasma at the initial stage of the analysis permits an accurate determination of the total tocopherol in RBC or plasma by calculations based on the recovery of the added isotope. This procedure for erythrocyte tocopherol analysis is compared with a gas-liquid chromatographic method in current use.
本文描述了一种相对快速的分光光度法，用于测定红细胞（RBC）中的总生育酚，该方法基于对原始Emmerie-Engel反应的改进。该方法的关键特征在于，在皂化和萃取阶段存在大量添加的抗氧化剂，如焦性没食子酸或抗坏血酸，以及使用薄层色谱法纯化生育酚。测定了许多人类受试者、无β脂蛋白血症患者和大鼠的血浆和红细胞中的总生育酚水平。结果发现，这些水平在正常人类受试者中范围很广，但红细胞与血浆生育酚的比率相对恒定，未校正时为0.18，当红细胞和血浆值均校正至100%回收率时为0.21。大鼠的红细胞与血浆比率为0.39。当通过体外和体内技术将放射性维生素E引入血液时，通过测量[(14)C]生育酚在红细胞和血浆中的分布，验证了分光光度法测定该比率的准确性。在分析初始阶段向红细胞或血浆中添加放射性生育酚，可通过基于添加同位素回收率的计算准确测定红细胞或血浆中的总生育酚。将这种红细胞生育酚分析方法与目前使用的气液色谱法进行了比较。
