采用单溴代联苯胺柱前衍生化结合反相高效液相色谱法测定硫辛酸。
Determination of lipoic acid by precolumn derivatization with monobromobimane and reversed-phase high-performance liquid chromatography.
作者信息
Witt W, Rüstow B
机构信息
Department of Neonatology, Hospital for Pediatrics, University Hospital Charité, Humboldt University at Berlin, Germany.
出版信息
J Chromatogr B Biomed Sci Appl. 1998 Jan 23;705(1):127-31. doi: 10.1016/s0378-4347(97)00488-x.
A new method for the quantitation of lipoic acid in plasma and tissues based on the selective precolumn derivatization of thiols with the fluorescent label monobromobimane is described. After extraction with diethyl ether, the dithiolane ring of lipoic acid is opened by reduction with NaBH4 before the free thiols can react with the label. Separation and quantitation was achieved by reversed-phase HPLC and fluorescence detection. The concentration-response curve was linear from 20 to 3000 nM in plasma. The recovery as determined with [3H]lipoic acid was 60.9% from plasma and 61.4% from rat heart tissue. The pretreatment of samples with N-ethylmaleimide makes it possible to differentiate between reduced and oxidized forms of lipoic acid.
描述了一种基于用荧光标记单溴联苯胺对硫醇进行选择性柱前衍生化来定量血浆和组织中硫辛酸的新方法。用乙醚萃取后，在游离硫醇与标记物反应之前，用NaBH4还原打开硫辛酸的二硫戊环。通过反相高效液相色谱和荧光检测实现分离和定量。血浆中浓度 - 响应曲线在20至3000 nM范围内呈线性。用[3H]硫辛酸测定的回收率在血浆中为60.9%，在大鼠心脏组织中为61.4%。用N - 乙基马来酰亚胺对样品进行预处理可以区分还原型和氧化型硫辛酸。
Zotero 插件