Ohisa Y, Kawamura T, Mizonobe K, Funato T, Sasaki T
Department of Clinical & Laboratory Medicine, Tohoku University School of Medicine, Sendai.
Rinsho Byori. 1995 Jun;43(6):630-6.
A chemiluminescence high performance liquid chromatography (HPLC) method for the simultaneous measurement of unsaturated fatty acid and lipid peroxide is newly described. This method was developed using reversed-phase HPLC which could separate unsaturated fatty acid and lipid peroxide clearly. Linear relationship was observed between these lipid peroxides and their peak areas on the chromatogram. The hydroperoxides were recently suggested to cause some diseases such as gastric lesions due to stress, but convenient method has not been established. The most popular method is the thiobarbituric acid (TBA) test, which has been extensively modified during the last several year. But the major disadvantage in this method is the interference of other thiobarbituric acid reactive substances (TBARS) and it is uncertain whether TBARS reflects the hydroperoxide concentration. This method is a highly sensitive fluorometric method for the quantitation of hydroperoxide at the nanomole level. Chemiluminescence was produced through luminol oxidation during the reaction of the hydroperoxide and cytochrome cheme. It is concluded that reversed phase HPLC can complement each other to serve as a direct and sensitive method for the determination of unsaturated fatty acid and lipid peroxide levels in vivo.
本文新描述了一种用于同时测定不饱和脂肪酸和脂质过氧化物的化学发光高效液相色谱（HPLC）方法。该方法采用反相HPLC开发，能够清晰分离不饱和脂肪酸和脂质过氧化物。在这些脂质过氧化物与其在色谱图上的峰面积之间观察到线性关系。最近有人提出氢过氧化物会引发一些疾病，如应激引起的胃部病变，但尚未建立便捷的检测方法。最常用的方法是硫代巴比妥酸（TBA）试验，在过去几年中该方法已被广泛改进。但该方法的主要缺点是其他硫代巴比妥酸反应性物质（TBARS）的干扰，并且不确定TBARS是否反映氢过氧化物浓度。此方法是一种用于在纳摩尔水平定量氢过氧化物的高灵敏度荧光法。在氢过氧化物与细胞色素c反应过程中，通过鲁米诺氧化产生化学发光。结论是反相HPLC可以相互补充，作为一种直接且灵敏的方法用于测定体内不饱和脂肪酸和脂质过氧化物水平。
