Peres Patrícia S, Valerio Andressa, Martinez Glaucia R
Laboratório de Oxidações Biológicas, Departamento de Bioquímica e Biologia Molecular, Setor de Ciências Biológicas, Universidade Federal do Paraná (UFPR), Curitiba, Brazil.
Biotechniques. 2016 Jun 1;60(6):279-84. doi: 10.2144/000114424. eCollection 2016.
The free nucleoside 2'-deoxyguanosine (dGuo) is the most susceptible to oxidation by reactive oxygen species (ROS) compared to the other free nucleosides, and its oxidation product 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo) has been used as a biomarker to quantify oxidative stress damage. We investigated different reactions using Fe(2+) or Cu(2+) and H2O2 in order to identify the reaction with the best yield. HPLC coupled with a UV detector and micrOTOF mass spectrometry were used to detect and confirm the identity of 8-oxodGuo. The optimized reaction synthesized 8-oxodGuo with a yield of 72.0%, much higher than that previously described in the literature. Our improved method for 8-oxodGuo synthesis could be extremely useful for assays that require the synthesis of internal standards labeled with stable isotopes.
与其他游离核苷相比，游离核苷2'-脱氧鸟苷（dGuo）最易被活性氧（ROS）氧化，其氧化产物8-氧代-7,8-二氢-2'-脱氧鸟苷（8-oxodGuo）已被用作量化氧化应激损伤的生物标志物。我们研究了使用Fe(2+)或Cu(2+)与H2O2的不同反应，以确定产率最高的反应。采用高效液相色谱结合紫外检测器和微量飞行时间质谱来检测和确认8-oxodGuo的身份。优化后的反应合成8-oxodGuo的产率为72.0%，远高于文献中先前描述的产率。我们改进的8-oxodGuo合成方法对于需要合成稳定同位素标记内标的分析可能极其有用。
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