Sanny C G, Rymas K
Oklahoma College of Osteopathic Medicine and Surgery, Tulsa 74107.
Anal Biochem. 1988 Jul;172(1):51-5. doi: 10.1016/0003-2697(88)90410-1.
Methods for separation of isoenzymes using high-performance (pressure) liquid chromatography have been adapted for analysis of aldehyde dehydrogenase (ALDH) (aldehyde:NAD oxidoreductase; EC 1.2.1.3) isoenzymes in canine liver. Two major peaks (peaks 1 and 3) and one minor peak (peak 2) of ALDH activity can be detected using ion-exchange chromatography and a continuous-flow analyzer. The elution profiles of ALDH activity in liver homogenates correspond with profiles obtained from isolated ALDH isoenzymes. Varying the composition of the ALDH assay reagents results in changes in the elution profiles consistent with the kinetic properties of the individual isoenzymes. Changes in elution profiles could be detected in liver samples following either in vitro or in vivo treatment with disulfiram (Antabuse). HPLC analysis may be a useful method to identify drug and genetic effects on ALDH isoenzymes and to investigate the mechanisms of altered aldehyde oxidation in animals.
利用高效（压力）液相色谱法分离同工酶的方法已被应用于犬肝脏中醛脱氢酶（ALDH，醛：NAD氧化还原酶；EC 1.2.1.3）同工酶的分析。使用离子交换色谱法和连续流动分析仪可检测到ALDH活性的两个主要峰（峰1和峰3）以及一个次要峰（峰2）。肝脏匀浆中ALDH活性的洗脱图谱与从分离的ALDH同工酶获得的图谱一致。改变ALDH测定试剂的组成会导致洗脱图谱发生变化，这与各个同工酶的动力学特性一致。在用双硫仑（戒酒硫）进行体外或体内处理后的肝脏样品中可检测到洗脱图谱的变化。高效液相色谱分析可能是一种有用的方法，可用于识别药物和基因对ALDH同工酶的影响，并研究动物体内醛氧化改变的机制。
