烟草科技2017，Vol.50Issue(5)：53-59,7.DOI:10.16135/j.issn1002-0861.2017.0017
UPLC-HRMS法定量分析小鼠组织中的6-氧-甲基鸟嘌呤
Quantitative analysis of 6-O-methylguanine in mouse tissues by UPLC-HRMS
摘要
Abstract
In order to evaluate the carcinogenicity of 4-(N-methyl-N-nitrosamino)-1-(3-pyridyl)-1-butanone (NNK), an ultra-high performance liquid chromatography-high resolution mass spectrometry (UPLC-HRMS) method was established to accurately determine the levels of 6-O-methylguanine(6-O-MeG)in mouse liver and lung tissues. The contents of 6-O-MeG in the liver and lung of normal saline-treated group, pyrazole-treated group and 5-methoxypsoralen (5-MOP)-treated group were measured separately after NNK administration. Tissue samples were extracted by DNA extraction kits, then the extraction was acidified at 80 ℃, followed by adding[CD3]6-O-MeG as the internal standard. After going through HLB solid phase column and redissolution by acetonitrile, the extraction was analyzed by UPLC-HRMS. The results showed that: 1) 6-O-MeG displayed good linearity in the range of 0.05-10.00 ng/mL with R2 of 0.9998, the limit of detection(LOD)of 0.011 ng/mL and the recoveries from 92.1% to 102.2% , the RSDs of intra- and inter-day were 1.10%-4.34% and 1.89%-6.58%, respectively. 2) The concentration of 6-O-MeG in the liver and lung of mice treated with pyrazole was 2.09 and 1.87 times as high as those treated with saline four hours after NNK administration;the 5-MOP-treated group was 0.32 and 0.67 times as high as the control group. The results indicated that cytochrome P4502A5 (CYP2A5) enzyme played an important role in the activation of NNK metabolism in mice. This method features a simple sample preparation, good selectivity, high sensitivity, and it is suitable for determining 6-O-MeG in mouse tissue.关键词
NNK/6-氧-甲基鸟嘌呤(6-O-MeG)/吡唑/5-甲氧基补骨脂素(5-MOP)/C57BL/6小鼠/超高效液相色谱-高分辨质谱(UPLC-HRMS)Key words
NNK/6-O-methylguanine (6-O-MeG)/Pyrazole/5-Methoxypsoralen(5-MOP)/C57BL/6 mouse/Ultra-high performance liquid chromatography-high resolution mass spectrometry (UPLC-HRMS)分类
轻工纺织引用本文复制引用
李琛,王荣浩,毛健,霍现宽,王丁众,张建勋..UPLC-HRMS法定量分析小鼠组织中的6-氧-甲基鸟嘌呤[J].烟草科技,2017,50(5):53-59,7.基金项目
国家自然科学基金面上项目"改性介孔材料固相萃取/UPLC-MSn方法研究烟草特有亚硝胺在动物体内代谢"(21175153) （21175153）
国家自然科学基金青年基金项目"烟碱在大鼠脑内代谢的活体动态分析及其神经递质化学效应的实时评价"(21307163). （21307163）
