High-performance liquid chromatography-tandem mass spectrometry measurement of radiation-induced base damage to isolated and cellular DNA
High-performance liquid chromatography-tandem mass spectrometry measurement of radiation-induced base damage to isolated and cellular DNA
复制标题
DOI：
10.1021/tx000085h
复制
发表时间：
2000-10-01
影响因子：
4.1
通讯作者：
Cadet, J
中科院分区：
文献类型：
--
作者：
Frelon, S;Douki, T;Cadet, J
建立了多反应监测模式下高效液相色谱（HPLC）分离与串联质谱（MS/MS）检测相结合的方法，用于评估辐射诱导的DNA碱基降解产物。这种灵敏且特异的检测方法旨在评估分离 DNA 和细胞 DNA 中的六种氧化 2'-脱氧核糖核苷和两种修饰的嘌呤碱基。为此，制备了稳定同位素标记的内标并用于同位素稀释质谱测量。后一种方法通过与其他两种测定方法的比较得到验证，包括与电化学检测相关的高效液相色谱法和与质谱联用的气相色谱法。使用特异性和灵敏的 HPLC-MS/MS 方法，5,6-二羟基-5,6-二氢胸苷、5-羟基-2'-脱氧尿苷、5-(羟甲基)-2'-脱氧尿苷、5-甲酰基-2'-脱氧尿苷、8-氧代-7,8-二氢-2'-脱氧腺苷、在暴露于 γ 辐射后，对分离的 DNA 和细胞 DNA 中的 8-氧代-7,8-二氢-2'-脱氧鸟苷、4,6-二氨基-5-甲酰胺嘧啶和 2,6-二氨基-4-羟基-5-甲酰胺嘧啶进行定量。
A method involving high-performance liquid chromatography (HPLC) separation associated with tandem mass spectrometry (MS/MS) detection in the multiple-reaction monitoring mode was set up for the assessment of radiation-induced degradation products of DNA bases. This sensitive and specific assay is aimed at assessing six oxidized 2'-deoxyribonucleosides and two modified purine bases within both isolated and cellular DNA. For this purpose, stable isotopically labeled internal standards were prepared and used for isotope dilution mass spectrometry measurements. The latter method was validated through a comparison with two other assays, including HPLC associated with electrochemical detection and gas chromatography coupled to mass spectrometry. Using the specific and sensitive HPLC-MS/MS approach, 5,6-dihydroxy-5,6-dihydrothymidine, 5-hydroxy-2'-deoxyuridine, 5-(hydroxymethyl)-2'-deoxyuridine, 5-formyl-2'-deoxyuridine, 8-oxo-7,8-dihydro-2'-deoxyadenosine, 8-oxo-7,8-dihydro-2'-deoxyguanosine, 4,6-diamino-5-formamidopyrimidine, and 2,6-diamino-4-hydroxy-5-formamidopyrimidine were quantified within both isolated and cellular DNA upon exposure to gamma -radiation.