Ultrasensitive Simultaneous Quantification of 1,N2-Etheno-2′-deoxyguanosine and 1,N2-Propano-2′-deoxyguanosine in DNA by an Online Liquid Chromatography-Electrospray Tandem Mass Spectrometry Assay
Ultrasensitive Simultaneous Quantification of 1,N2-Etheno-2′-deoxyguanosine and 1,N2-Propano-2′-deoxyguanosine in DNA by an Online Liquid Chromatography-Electrospray Tandem Mass Spectrometry Assay
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DOI：
10.1021/tx1001018
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发表时间：
2010-07-01
影响因子：
4.1
通讯作者：
Medeiros, Marisa H. G.
中科院分区：
文献类型：
--
作者：
Garcia, Camila C. M.;Freitas, Florencio P.;Medeiros, Marisa H. G.
外源性和内源性化合物产生的外环DNA加合物正在成为研究各种人类疾病和空气污染暴露的潜在工具。建立了一种在线反相高效液相色谱-电喷雾串联质谱检测、多反应监测模式和稳定同位素标记内标同时定量DNA中1，n -2-乙烯-2′-脱氧鸟苷（1,n -2-epsilon dGuo）和1，n -2-丙烷-2′-脱氧鸟苷（1,n -2- propanodguo）的高灵敏度方法。该方法允许在100 μ g水解DNA中直接在线定量2'-脱氧鸟苷和约500 amol的加合物。利用新技术，准确测定了人培养细胞(4.01 +/- 0.32 1,N-2-epsilon dGuo/10（8) dGuo和3.43 +/- 0.33 1,N-2-propanodGuo/10(8) dGuo）和大鼠组织（肝脏，2.47 +/- 0.61 1,N-2-epsilon dGuo/10(8) dGuo和4.61 +/- 0.69 1,N-2-propanodGuo/108 dGuo） DNA提取物中1，n -2-乙烯-2'-脱氧鸟苷和1，n2 - propanod2 '-脱氧鸟苷的含量；脑，2.96 +/- 1.43,N-2-epsilon dGuo/10(8) dGuo和5.66 +/- 3.70 1,N-2-propanoclGuo/10(8) dGuo；肺分别为0.87 +/- 0.34 1,N-2-edGuo/ 10(8) dGuo和2.25 +/- 1.72 1,N-2-propanodGuo/10(8) dGuo。本文所述的方法可以通过对人体和病理条件下及空气污染暴露后不同加合物的定量研究外环DNA加合物的生物学意义。
Exocyclic DNA adducts produced by exogenous and endogenous compounds are emerging as potential tools to study a variety of human diseases and air pollution exposure. A highly sensitive method involving online reverse-phase high performance liquid chromatography with electrospray tandem mass spectrometry detection in the multiple reaction monitoring mode and employing stable isotope-labeled internal standards was developed for the simultaneous quantification of 1,N-2-etheno-2'-deoxyguanosine (1,N-2-epsilon dGuo) and 1,N-2-propano-2'-deoxyguanosine (1,N-2-propanodGuo) in DNA. This methodology permits direct online quantification of 2'-deoxyguanosine and ca. 500 amol of adducts in 100 mu g of hydrolyzed DNA M the same analysis. Using the newly developed technique, accurate determinations of 1,N-2-etheno-2'-deoxyguanosine and 1,N2-propano-2'-deoxyguanosine levels in DNA extracts of human cultured cells (4.01 +/- 0.32 1,N-2-epsilon dGuo/10(8) dGuo and 3.43 +/- 0.33 1,N-2-propanodGuo/10(8) dGuo) and rat tissue (liver, 2.47 +/- 0.61 1,N-2-epsilon dGuo/10(8) dGuo and 4.61 +/- 0.69 1,N-2-propanodGuo/108 dGuo; brain, 2.96 +/- 1.43,N-2-epsilon dGuo/10(8) dGuo and 5.66 +/- 3.70 1,N-2-propanoclGuo/10(8) dGuo; and lung, 0,87 +/- 0.34 1,N-2-edGuo/ 10(8) dGuo and 2.25 +/- 1.72 1,N-2-propanodGuo/10(8) dGuo) were performed. The method described herein can be used to study the biological significance of exocyclic DNA adducts through the quantification of different adducts in humans and experimental an with pathological conditions and after air pollution exposure.