Are HPLC and the comet assay equally good at detecting 8-oxoGua?
Figure 1 illustrates two possible explanations for the discrepant background determinations that have been reported for comet assay and HPLC methods. A difference in dose–response slopes between HPLC and the comet assay would suggest differing abilities to detect 8-oxoGua. On the other hand, similar slopes would indicate that oxidation during sample preparation is the main cause of these differences. For the dose–response curve obtained when cellular DNA is damaged with Ro 19-8022 and light, the comet assay gives a slope apparently slightly greater than that for HPLC analysis, but they are not significantly different (P > 0.5). FPG is not specific to 8-oxoGua, but detects also ring-opened purines (fapy bases) (11). If there are significant numbers of fapy derivatives in the background damage, or in the damage induced by Ro 19-8022 plus light, FPG-sensitive sites as measured with the comet assay would be an over-estimate of 8-oxoGua content. Pflaum et al. (7) found that, in phage DNA tr
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