Protocol for the simultaneous quantification of oxidative purine lesions in DNA using LC-MS/MS analysis
Chryssostomos Chatgilialoglu1, Marios G Krokidis2, Michael A Terzidis3
1Institute for Organic Synthesis and Photoreactivity, National Research Council, 40129 Bologna, Italy; Center of Advanced Technologies, Adam Mickiewicz University, 61-712 Poznań, Poland.
None:
Most DNA damages induced through oxidative metabolism are single lesions which can accumulate in tissues. Here, we present a protocol for the simultaneous quantification of oxidative purine lesions (cPu and 8-oxo-Pu) in DNA. We describe steps for enzymatic digestion of DNA and sample pre-purification, followed by quantification through liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. We optimized this protocol in commercially available calf thymus DNA and used genomic and mitochondrial DNA extracted from cell cultures and animal and human tissues.
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