高效液相色谱法测定红细胞内α-生育酚

High performance liquid chromatography measuring α generational hydroxybenzene in red blood cell

目的 :寻找一种测定红细胞内α 生育酚含量的较好方法。方法 :将含有α 生育酚的肝素抗凝血离心后吸出血浆 ,剩余的红细胞用生理盐水洗涤后用氢氧化钾皂化 ,并加入联苯三酚以防止α 生育酚被氧化。再用正己烷提取红细胞内的α 生育酚 ,并进行高效液相层析。结果 :α 生育酚浓度为 0 92 7～ 9 2 6 7μmol/L ,线性关系良好 ,最低检测限为 0 32 3μmol/L ,变异系数 1 3% ,回收率 1 0 0 2 %。 结论 :该方法简单可靠 ,重复性好 ,并可从细胞水平上了解机体的抗脂质过氧化作用。

Aim:To find a better method to measure α generational hydroxybenzene in red blood cell (RBC).Methods:The α generational hydroxybenzene in RBC was measured with high performance liquid chromatography (HPLC).After the heparinizing anticoagulating blood containing α generational hydroxybenzene was centrifuged and plasma was exsuctioned,the residual RBC was lavaged with physiological salt solution,then saponified with potassium hydroxide,while added diphenyl triester to prevent α generational hydroxybenzene oxidized.After those,α generational hydroxybenzene in RBC was extracted with positive ethane,then isolated by HPLC.Results:The linear relation was well,while the concentration of α generational hydroxybenzene was between 0.927 ～9.267 μmol/L.The lowest limit of detectable level of α generational hydroxybenzene was 0.323 μmol/L,the variable coefficient was 1.3%,and the rate of recovery was 100.2%.Conclusion:The technique was simple and reliable,better repetitive,and helped to understand anti hyperoxide lipid of human body in cellular level.