Stalikas C D, Konidari C N
Department of Chemistry, University of Ioannina, Ioannina, 45 110, Greece.
Anal Biochem. 2001 Mar 1;290(1):108-15. doi: 10.1006/abio.2000.4951.
A gas chromatographic method is described for the quantification of free and total malondialdehyde (MDA) in biological materials. The procedure involves derivatization of the analyte with 2,4,6-trichlorophenylhydrazine, extraction with n-hexane, and separation of the cyclic derivatization product on a OV-5 gas chromatographic column. Concentration of the derivatization reagent, pH, reaction time, and temperature were investigated to determine the optimal derivatization conditions. Under these conditions, the method allows for the selective detection of free and total MDA at femtomole levels in several biological materials without any interferences. The procedure yields relative standard deviation values for the intra- and interassays in the range 3.3 and 3.9%, respectively, for the electron-capture and mass-selective (SIM mode) detection systems. Recoveries of MDA from spiked matrices reached 96%. The present method offers the advantage of the alternative use of either electron-capture or mass-selective detection. Furthermore it avoids overestimation of MDA since it employs mild conditions for sample processing and there is no need for preventing protein separation for the assessment of free MDA.
本文描述了一种用于定量生物材料中游离和总丙二醛（MDA）的气相色谱方法。该程序包括用2,4,6 - 三氯苯肼对分析物进行衍生化、用正己烷萃取，以及在OV - 5气相色谱柱上分离环状衍生化产物。研究了衍生化试剂的浓度、pH值、反应时间和温度，以确定最佳衍生化条件。在这些条件下，该方法能够在几种生物材料中以飞摩尔水平选择性检测游离和总MDA，且无任何干扰。对于电子捕获和质量选择（SIM模式）检测系统，该程序的批内和批间相对标准偏差值分别在3.3%和3.9%范围内。加标基质中MDA的回收率达到96%。本方法具有可交替使用电子捕获或质量选择检测的优点。此外，由于它采用温和的样品处理条件，且无需为评估游离MDA而防止蛋白质分离，因此避免了对MDA的高估。
