SIMULTANEOUS QUANTITATION OF OXIDIZED AND REDUCED GLUTATHIONE IN EQUINE BIOLOGICAL-FLUIDS BY REVERSED-PHASE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY USING ELECTROCHEMICAL DETECTION
SIMULTANEOUS QUANTITATION OF OXIDIZED AND REDUCED GLUTATHIONE IN EQUINE BIOLOGICAL-FLUIDS BY REVERSED-PHASE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY USING ELECTROCHEMICAL DETECTION
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DOI：
10.1016/0378-4347(95)00245-e
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发表时间：
1995-11-03
期刊：
影响因子：
--
通讯作者：
MILLS, PC
中科院分区：
文献类型：
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作者：
SMITH, NC;DUNNETT, M;MILLS, PC
还原型谷胱甘肽 (GSH) 与氧化型谷胱甘肽 (GSSG) 比率的变化可用于指示体内氧化应激。开发了一种使用库仑电化学检测 (LCEC) 的快速、高灵敏度等度反相高效液相色谱方法，可同时检测马生物体液中的 GSH 和 GSSG。使用高氯酸从马血浆和溶血液中提取 GSH 和 GSSG，使用甲醇对支气管肺泡灌洗液样品进行脱蛋白。将提取物注射到 Hypersil ODS HPLC 柱上，产生与 GSH 和 GSSG 相对应的高分辨率峰。在马溶血物中发现的 GSH 和 GSSG 浓度与之前在人类和实验动物中发现的浓度相似，尽管据作者所知，之前使用 LCEC 测量支气管肺泡灌洗液中 GSH 和 GSSG 的尝试并未成功。该方法可用于测量在运动和疾病等可能诱发氧化应激的生理条件下生物体液中的 GSH 氧化还原比。
A change in the ratio of reduced glutathione (GSH) to oxidized glutathione (GSSG) can be used to indicate oxidative stress in vivo. A rapid and highly sensitive isocratic reversed-phase high-performance liquid chromatographic method using coulometric electrochemical detection (LCEC) has been developed to simultaneously detect GSH and GSSG in equine biological fluids. Perchloric acid was used to extract GSH and GSSG from equine plasma and haemolysates, and methanol was used to deproteinise bronchoalveolar lavage fluid samples. Injection of extracts onto a Hypersil ODS HPLC column produced well resolved peaks corresponding to GSH and GSSG. The concentrations of GSH and GSSG found in equine haemolysates were similar to those previously found in humans and laboratory animals, although, to the authors' knowledge, previous attempts to measure GSH and GSSG in bronchoalveolar lavage fluid using LCEC have been unsuccessful. This method can be used to measure the GSH redox ratio in biological fluids during physiological conditions that may induce oxidative stress, such as exercise and disease.