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damage to DNA is considered to be important in mutagenesis, carcinogenesis and the ageing process. 1,2 Of about 20 major oxidative DNA adducts that have been characterized, 3 8-hydroxy-2′-deoxyguanosine (8OHdG) has received considerable attention due to its demonstrated mutagenic potential, 1,4,5 which has been shown to cause the GC→TA transversion. 6,7 It is worth mentioning that many GC→TA transversions have been detected in p 53 genes (an important carcinogenic gene) isolated from human lung and liver cancers. 8 This result suggests that 8OHdG may be a good marker of carcinogenesis. Since oxidative damage to DNA is a continuous process, intracellular repair mechanisms have evolved to avoid the rapid and lethal accumulation of oxidative DNA damages. In the case of 8OHdG, the experimental evidence indicates that mammalian cells possess nonspecific (excinucleases) and specific (glycosylases) DNA repair enzymes that can excise this modification, which is subsequently excreted in urine as nucleoside or free base without any further metabolism. 9–11 The determination of urinary 8OHdG has thus been proposed as a noninvasive assay of in vivo oxidative DNA lesions. Loft et al. have reported that smokers excrete 50% more urinary 8OHdG than nonsmokers. This indicates a 50% increased rate of oxidative DNA damage from smoking. 12 The urinary 8OHdG levels in patients with gynecologic cancer 13 and non-insulin-dependent diabetes mellitus 14 are also significantly higher than that in control subjects. At present, several methodologies for measuring 8OHdG have been developed, including high-performance liquid chromatography (HPLC) with electrochemical (EC) detection and gas chromatography/mass spectrometry (GC/MS). Although HPLC-EC is a highly sensitive method for the analysis of 8OHdG, the applicability of this method is not as general as that expected from a suitable and sensitive GC/MS method. To the best of our knowledge, there have been a few reports concerning the analysis of urinary 8OHdG by GC/MS. 15,16 However, for a method before GC analysis of the products of oxidative DNA damage can be accomplished, these compounds must first be converted to volatile derivatives with suitable chromatographic properties. Among various derivatives of nucleoside, the trimethylsilyl (TMS) derivative is the most suitable to work for the derivatization of nucleosides when compared to other derivatives. Although there are a few reports of studied concerning the best reaction condition for the derivatization of normal nucleosides with TMS reagents, 17,18 these investigations were carried out at the milligram level. Here, we report on an optimization of the derivatization procedure …

Method for the Analysis of 8-Hydroxy-2′-deoxyguanosine in Urine by Gas Chromatography