Shigenaga M K, Park J W, Cundy K C, Gimeno C J, Ames B N
Methods Enzymol. 1990;186:521-30. doi: 10.1016/0076-6879(90)86146-m.
HPLC with electrochemical detection is a highly sensitive and selective method for detecting the oxidatively modified DNA residue oh8dG. By this method, the detection of oh8dG from DNA and urine offers a powerful approach for assessing in vivo oxidative damage. Application of this technique to the detection of oh8dG from DNA permits the quantitation of the steady-state levels of this oxidatively modified deoxynucleoside and overcomes the detection problems associated with the extremely low levels present in DNA. In addition, the selectivity gained by this detection method eliminates the problem of separating the signal for oh8dG from normal deoxynucleosides. The quantitation of oh8dG in urine complements the measurement of oh8dG in DNA by estimating cumulative oxidative DNA damage in the body. In addition, the urinary assay provides a noninvasive means of measuring this type of damage in laboratory animals and human populations. Thus, an individual animal or human subject may be monitored over time, possibly under various prooxidant conditions, using oh8dG as a sensitive marker for oxidative DNA damage. This analytical approach may allow one to estimate the exposure of an individual to prooxidant conditions associated with lifestyle, genetic predisposition, degenerative diseases, and environmental toxins.
高效液相色谱-电化学检测法是一种检测氧化修饰的DNA残基oh8dG的高灵敏度和高选择性方法。通过这种方法，从DNA和尿液中检测oh8dG为评估体内氧化损伤提供了一种有力的途径。将该技术应用于从DNA中检测oh8dG，可以对这种氧化修饰的脱氧核苷的稳态水平进行定量，并克服了与DNA中极低水平相关的检测问题。此外，这种检测方法所获得的选择性消除了将oh8dG信号与正常脱氧核苷信号分离的问题。尿液中oh8dG的定量通过估计体内累积的氧化DNA损伤，补充了DNA中oh8dG的测量。此外，尿液检测为在实验动物和人群中测量这类损伤提供了一种非侵入性手段。因此，使用oh8dG作为氧化DNA损伤的敏感标志物，可以对单个动物或人类受试者进行长期监测，可能是在各种促氧化剂条件下。这种分析方法可能使人们能够估计个体暴露于与生活方式、遗传易感性、退行性疾病和环境毒素相关的促氧化剂条件的情况。
