Chemical modification of DNA: quantitation of O6-methyldeoxyguanosine by tandem mass spectrometry.
Chemical modification of DNA: quantitation of O6-methyldeoxyguanosine by tandem mass spectrometry.
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DNA 的化学修饰：通过串联质谱法定量 O6-甲基脱氧鸟苷。
DOI：
10.1002/bms.1200200812
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发表时间：
1991
期刊：
影响因子：
--
通讯作者：
Cooks,RG
中科院分区：
文献类型：
--
作者：
Chae,WG;Chang,CJ;Wood,JM;Cooks,RG
本文描述了一种定量的方法，该方法是由甲基甲烷磺酸盐（MeMS）体外甲基化小牛胸腺DNA产生的o6 -甲基脱氧鸟苷。当DNA用弱致癌物MeMS处理时，以0.02%的产率（表示为o6 -甲基脱氧鸟苷与脱氧鸟苷）生成10 (-11)-10 (-12)mol的样品，定量精度达到10%。这些结果显示了该方法在分析与许多烷基化剂诱导生物效应相关的低水平DNA化学修饰方面的潜在应用。该方法利用酶降解，反相色谱法，最后通过解吸化学电离串联质谱法进行分析。多重反应监测用于提高灵敏度，cd3标记核苷作为内标进行定量。
Methodology is described which allows quantitation of O6-methyldeoxyguanosine generated as a product of in vitro methylation of calf thymus DNA by methyl methanesulfonate (MeMS). Quantitative precision of 10% is achieved on samples of 10 (-11)-10 (-12) mol generated in 0.02% yield (expressed as O6-methyldeoxyguanosine versus deoxyguanosine) when DNA is treated with the weak carcinogen MeMS. These results show the potential application of this method to the analysis of DNA chemical modifications at the low levels that are relevant to the induction of biological effects of many alkylating agents. The methodology utilizes enzymatic degradation, reverse-phase chromatography and finally analysis by tandem mass spectrometry using desorption chemical ionization. Multiple reaction monitoring was used to increase sensitivity and the CD3-labeled nucleoside was used as an internal standard for quantification.