Müller M, Belas F J, Blair I A, Guengerich F P
Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, Tennessee 37232, USA.
Chem Res Toxicol. 1997 Feb;10(2):242-7. doi: 10.1021/tx960124i.
High performance liquid chromatography (HPLC)/electrospray mass spectrometry methods were developed for the analysis of 1,N2-etheno(epsilon)guanine (Gua) and 5,6,7,9-tetrahydro-7-hydroxy-9-oxoidmidazo[1,2-a]purine (HO-ethanoGua) [the cyclized form of N2-(2-oxoethyl)Gua] and its deoxyribose derivative in DNA. Evidence was provided for the formation of the latter adduct in DNA treated with 2-chlorooxirane, the reactive product formed from the carcinogen vinyl chloride. Measured levels of HO-ethanoGua and HO-ethanodeoxyguanosine were similar, although the assay for the deoxyribosyl derivative has some technical advantages. 3,N4-epsilon-Deoxycytidine was also estimated in 2-chlorooxirane-treated DNA using HPLC with fluorescence detection. Levels of all known adducts formed from vinyl chloride have now been estimated in DNA treated with 2-chlorooxirane and vary in the order N7-(2-oxoethyl)Gua >> 1,N6-epsilon-adenine > HO-ethanoGua > N2,3-epsilon-Gua > 3,N4-epsilon-cytosine > 1,N2-epsilon-Gua. Although in vivo adduct levels may not parallel these due to differential stability and rates of repair, analyses of the adducts in DNA treated with 2-chlorooxirane provide a basis for consideration of the biological effects of these adducts.
已开发出高效液相色谱（HPLC）/电喷雾质谱法，用于分析DNA中的1,N2-乙烯基鸟嘌呤（Gua）、5,6,7,9-四氢-7-羟基-9-氧代咪唑并[1,2-a]嘌呤（HO-乙烯基鸟嘌呤）[N2-(2-氧代乙基)鸟嘌呤的环化形式]及其脱氧核糖衍生物。在用2-氯环氧乙烷（由致癌物氯乙烯形成的反应产物）处理的DNA中，为后一种加合物的形成提供了证据。尽管对脱氧核糖基衍生物的检测具有一些技术优势，但测得的HO-乙烯基鸟嘌呤和HO-乙烯基脱氧鸟苷水平相似。还使用带荧光检测的HPLC对2-氯环氧乙烷处理的DNA中的3,N4-ε-脱氧胞苷进行了估算。现在已对用2-氯环氧乙烷处理的DNA中由氯乙烯形成的所有已知加合物的水平进行了估算，其顺序为N7-(2-氧代乙基)鸟嘌呤>>1,N6-ε-腺嘌呤>HO-乙烯基鸟嘌呤>N2,3-ε-鸟嘌呤>3,N4-ε-胞嘧啶>1,N2-ε-鸟嘌呤。尽管由于稳定性和修复速率不同，体内加合物水平可能与此不平行，但对用2-氯环氧乙烷处理的DNA中的加合物进行分析为考虑这些加合物的生物学效应提供了依据。
