Bogdanov M B, Beal M F, McCabe D R, Griffin R M, Matson W R
Neurochemistry Laboratory, Neurology Service, Massachusetts General Hospital and Harvard Medical School, Boston 02114, USA.
Free Radic Biol Med. 1999 Sep;27(5-6):647-66. doi: 10.1016/s0891-5849(99)00113-6.
8-Hydroxy-2'-deoxyguanosine (8OH2'dG) is a principal stable marker of hydroxyl radical damage to DNA. It has been related to a wide variety of disorders and environmental insults, and has been proposed as a useful systematic marker of oxidative stress. Analytic procedures for 8OH2'dG in DNA digests are well established; however, routine measurement of free 8OH2'dG in other body fluids such as urine or plasma has been problematic. This has hindered its evaluation as a general clinical, therapeutic monitoring, or environmental assessment tool. Therefore, we developed a liquid chromatography electrochemical column-switching system based on the use of the unique purine selectivity of porous carbon columns that allows routine accurate measurement of 8OH2'dG in a variety of biologic matrices. This paper describes the rationale of the system design and the protocols developed for 8OH2'dG in urine, plasma, cerebrospinal fluid, tissue, DNA, saliva, sweat, kidney dialysis fluid, foods, feces, culture matrix, and microdialysates. Concentrations in both human and animal body fluids and tissues are reported. The system performance is discussed in the context of a 1-year evaluation of the methods applied to approximately 3600 samples, using internal quality control and external blind testing to determine long-term accuracy. The methods are reliable and accurate, and therefore should prove useful in assessing the role and utility of oxidative DNA damage in aging and human illness.
8-羟基-2'-脱氧鸟苷（8OH2'dG）是DNA受到羟基自由基损伤的主要稳定标志物。它与多种疾病和环境损伤有关，并被提议作为氧化应激的一种有用的系统性标志物。DNA消化物中8OH2'dG的分析方法已经很成熟；然而，在尿液或血浆等其他体液中常规测量游离8OH2'dG一直存在问题。这阻碍了它作为一种通用的临床、治疗监测或环境评估工具的评估。因此，我们基于多孔碳柱独特的嘌呤选择性开发了一种液相色谱电化学柱切换系统，该系统能够对多种生物基质中的8OH2'dG进行常规准确测量。本文描述了该系统设计的原理以及针对尿液、血浆、脑脊液、组织、DNA、唾液、汗液、肾透析液、食物、粪便、培养基质和微透析液中8OH2'dG所制定的方案。报告了人体和动物体液及组织中的浓度。在对大约3600个样本应用这些方法进行为期1年的评估过程中，通过内部质量控制和外部盲法检测来确定长期准确性，进而对系统性能进行了讨论。这些方法可靠且准确，因此在评估氧化DNA损伤在衰老和人类疾病中的作用及效用方面应会被证明是有用的。
