Development for separation analysis of micro free radical materials and its application for disease elucidation

自由基在体内的作用与各种疾病的关系已经被研究，并且检测方法吸引了各个医学领域的广泛关注。电子自旋共振（ESR）谱已成功地检测和鉴定了各种化学和生物体系中的多种自由基。羧酸是重要的营养基质和代谢产物在活的生物体。本文研究了苯甲酰甲基溴与羧酸衍生的苯甲酰甲基酯在碱性条件下产生的自由基，并采用高效液相色谱法测定了对硝基苯甲酰甲基溴与羧酸衍生的对硝基苯甲酰甲基酯。对硝基苯甲酰甲基溴被用作柱前试剂以衍生羧酸。对硝基苯甲酰甲基羧酸酯经碱水解产生自由基，用电子自旋共振（ESR）和电子捕获检测器（ECD）检测，乙酸在10-500 pmol，丙酸在50-500 pmol，丁酸在50-1000 pmol，采用ESR自由基色谱法，以HPLC为检测器，同时检测了g值相近的自旋加合物所产生的自由基。

The roles of free radicals in the body have been investigated in the relationship with various diseases and the detection methods are attracting much attention from various medical fields. Many kind of free radicals in various chemical and biological systems have been successfully detected and identified by an electron spin resonance (ESR) spectroscopy. Carboxylic acids are important as nutritional substrates and metabolites in living organisms. It was found that phenacyl esters, which are derived from phenacyl bromide and carboxyl acids, give rise to the free radicals under the alkaline condition, The p-nitrophenacyl esters derived from p-nitrophenacyl bromide and carboxylic acids were determined by a high-performance liquid chromatography with ESR and ECD as detection. The p-nitrophenacyl bromide was used as a pre-column reagent to derivative the carboxylic acids. The p-nitrophenacyl esters of carboxylic acid gave free radicals by hydrolysis with an alkaline and then the free radicals were detected with ESR and ECD.The ECD peak height was related to the concentration with high linearity in the range of 10-500 pmol for acetic acid, 50-500 pmol for propionic acid, 50-1000 pmol for butyric acid, 0.1-1nmol for valeric and 0.1- 1nmol for hexanoic acid.The radical chromatography using ESR for HPLC detector was carried out for simultaneous detection of free radicals derived from similar spin adducts of g value.