Guetens Gunther, De Boeck Gert, Highley Martin, van Oosterom Allan T, de Bruijn Ernst A
Department of Chemistry, University of Antwerp, Belgium.
Crit Rev Clin Lab Sci. 2002 Sep;39(4-5):331-457. doi: 10.1080/10408360290795547.
All forms of aerobic life are subjected constantly to oxidant pressure from molecular oxygen and also reactive oxygen species (ROS), produced during the biochemical utilization of O2 and prooxidant stimulation of O2 metabolism. ROS are thought to influence the development of human cancer and more than 50 other human diseases. To prevent oxidative DNA damage (protection) or to reverse damage, thereby preventing mutagenesis and cancer (repair), the aerobic cell possesses antioxidant defense systems and DNA repair mechanisms. During the last 20 years, many analytical techniques have been developed to monitor oxidative DNA base damage. High-performance liquid chromatography-electrochemical detection and gas chromatography-mass spectrometry are the two pioneering contributions to the field. Currently, the arsenal of methods available include the promising high-performance liquid chromatography-tandem mass spectrometry technique, capillary electrophoresis, 32P-postlabeling, fluorescence postlabeling, 3H-postlabeling, antibody-base immunoassays, and assays involving the use of DNA repair glycosylases such as the comet assay, the alkaline elution assay, and the alkaline unwinding method. Recently, the use of liquid chromatography-mass spectrometry has been introduced for the measurement of a number of modified nucleosides in oxidatively damaged DNA. The bulk of available chromatographic methods aimed at measuring individual DNA base lesions require either chemical hydrolysis or enzymatic digestion of oxidized DNA, following extraction from cells or tissues. The effect of experimental conditions (DNA isolation, hydrolysis, and/or derivatization) on the levels of oxidatively modified bases in DNA is enormous and has been studied intensively in the last 10 years.
所有需氧生物都不断受到来自分子氧以及活性氧（ROS）的氧化压力，ROS是在O2的生化利用过程中产生的，并且会刺激O2代谢产生促氧化剂。ROS被认为会影响人类癌症以及其他50多种人类疾病的发展。为了防止氧化性DNA损伤（保护）或逆转损伤，从而防止诱变和癌症（修复），需氧细胞拥有抗氧化防御系统和DNA修复机制。在过去的20年里，已经开发出许多分析技术来监测氧化性DNA碱基损伤。高效液相色谱 - 电化学检测和气相色谱 - 质谱是该领域的两项开创性贡献。目前，可用的方法包括有前景的高效液相色谱 - 串联质谱技术、毛细管电泳、32P后标记、荧光后标记、3H后标记、抗体 - 碱基免疫测定以及涉及使用DNA修复糖基化酶的测定，如彗星试验、碱性洗脱试验和碱性解旋法。最近，液相色谱 - 质谱已被用于测量氧化损伤DNA中的多种修饰核苷。大多数旨在测量单个DNA碱基损伤的色谱方法需要在从细胞或组织中提取后对氧化DNA进行化学水解或酶消化。实验条件（DNA分离、水解和/或衍生化）对DNA中氧化修饰碱基水平的影响非常大，并且在过去10年中已进行了深入研究。
