干旱区生态环境知识资源中心(Arid): 液相色谱-串联质谱法同时检测DNA中3-甲基腺嘌呤和3-乙基腺嘌呤

Arid

	液相色谱-串联质谱法同时检测DNA中3-甲基腺嘌呤和3-乙基腺嘌呤
其他题名	Simultaneous Determination of N3-methyladenine and N3-ethyladenine in DNA by Liquid Chromatography-Tandem Mass Spectrometry
	田永峰 1; 侯宏卫 1; 张小涛 1; 刘勇 2; 王安 2; 胡清源 1
来源期刊	分析化学
ISSN	0253-3820
出版年	2014
卷号	42 期号:8 页码:1200-1204
中文摘要	利用阳离子交换固相萃取柱(Waters Oasis MCX)富集净化 DNA样品,建立了液相色谱串联质谱(LC-MS/MS)同时检测DNA中3-甲基腺嘌呤(N3-MeA)和3-乙基腺嘌呤(N3-EtA)的方法。采用氘代-3-甲基腺嘌呤(d3-N3-MeA)和氘代-3-乙基腺嘌呤(d5-N3-EtA)为内标;进样量3muL,分析时间为13 min;亲水相互作用色谱柱(Waters XBridge HILIC)进行液相分离,流动相为10 mmol/L甲酸铵-乙腈溶液(5:95, V/V, pH=4.0),流速250muL/min;质谱条件:电喷雾离子源,多反应监测正离子扫描方式;电喷雾电压:5500 V,雾化气:369 Pa,气帘气:185 Pa,电离温度:400℃,驻留时间:40 ms。本方法对N3-MeA和N3-EtA的检出限分别为0.043和0.007mug/L,方法回收率为87.8%~103.0%。采用本方法检测了卷烟烟气粒相物暴露的DNA中N3-MeA和N3-EtA含量。结果表明,卷烟烟气粒相物暴露后的小牛胸腺DNA中3-甲基腺嘌呤和3-乙基腺嘌呤可被本方法定量检出。
英文摘要	A liquid chromatography-tandem mass spectrometry (LC-MSMS) method has been developed for the simultaneous determination of N3-methyladenine (N3-MeA) and N3-ethyladenine (N3-EtA) in calf thymus DNA. The DNA samples has been purified and enriched by cation exchange cartridge (Waters Oasis MCX). d3-N3-MeA and d5-N3-EtA were used as isotope internal standard. The DNA samples were injected with autosampler. The injected volume was 3 muL and analysis time was 13 min. The sample separation was carried out on hydrophilic interaction chromatograph (Waters XBridge HILIC) with 10 mmol/L ammonium formate-acetonitrile (5:92, V/V, pH=4. 0) as mobile phase. The flow rate was set at 250 muL/min. Mass spectrometry was performed by electrospray ionization (ESI) with multi-reactions monitoring (MRM). The optimized operation conditions of MS were as follows: nebulizer gas 369 Pa; curtain gas 185 Pa, turbo ionspray temperature 400 ℃, ionspray voltage 5500 V, dwell time 40 ms. The limits of detection were 0. 043 and 0. 007 mug/L for N3-MeA and N3-EtA, respectively. The recoveries were between 87. 8% and 103. 0%for N3-MeA and N3-EtA. This method was successfully applied to the determination of N3-MeA and N3-EtA in calf thymus DNA by cigarette smoke condensate (CSC) exposure. This method is appropriate for routine analysis and accurate quantification of N3-MeA and N3-EtA by CSC exposure.
中文关键词	3-甲基腺嘌呤 ; 3-乙基腺嘌呤 ; 液相色谱串联质谱
英文关键词	DNA N3-Methyladenine N3-Ethyladenine Liquid chromatography-tandem mass spectrometry Deoxyribonucleic acid
语种	中文
国家	中国
收录类别	CSCD
WOS类目	CHEMISTRY MULTIDISCIPLINARY
WOS研究方向	Chemistry
CSCD记录号	CSCD:5215384
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/231039
作者单位	1.国家烟草质量监督检验中心, 郑州, 河南 450001, 中国; 2.中国科学院安徽光学精密机械研究所, 合肥, 安徽 230001, 中国
推荐引用方式 GB/T 7714	田永峰,侯宏卫,张小涛,等. 液相色谱-串联质谱法同时检测DNA中3-甲基腺嘌呤和3-乙基腺嘌呤[J],2014,42(8):1200-1204.
APA	田永峰,侯宏卫,张小涛,刘勇,王安,&胡清源.(2014).液相色谱-串联质谱法同时检测DNA中3-甲基腺嘌呤和3-乙基腺嘌呤.分析化学,42(8),1200-1204.
MLA	田永峰,et al."液相色谱-串联质谱法同时检测DNA中3-甲基腺嘌呤和3-乙基腺嘌呤".分析化学 42.8(2014):1200-1204.