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Improved preparation and identification of aristolochic acid-DNA adducts by solid-phase extraction with liquid chromatography-tandem mass spectrometry

查看全文 作　　者：GAO [1,2]Haiyan;GUO [2]Fangqiu;FENG [1]Feng;YIN [1]Junfa;SONG [1]Maoyong;WANG [1]Hailin 高影响力作者 机构地区：[1]State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China;[2]Research Center of Modernization of Chinese Medicines, Central South University, Changsha 410083, China高影响力机构 出　　处：《Journal of Environmental Sciences》索引2009年第21卷第12期,共8页高影响力期刊 基　　金：supported by the National Basic Research Program (973) of China (No. 2007CB407305,2008CB417201);the National High Technology Research and Development Program (863) of China (No.2007AA06A407);the National Natural Science Foundation of China (No. 20737003, 20621703, 20805057) 摘　　要：Aristolochic acid (AA) is a known nephrotoxin and potential carcinogen, which can form covalent DNA adducts after metabolic activation in vivo and in vitro. A simple method for preparation and characterization of aristolochic acid-DNA adducts was developed. Four AA-adducts were synthesized by a direct reaction of AAI/AAII with 2'-deoxynucleosides. The reaction mixture was first cleaned-up and pre-concentrated using solid phase extraction (SPE), and further purified by a reversed-phase high performance liquid chromatography (HPLC). By the application of developed SPE procedure, matrices and byproducts in reaction mixture could be greatly reduced and adducts of high purity (more than 94% as indicated by HPLC) were obtained. The purified AA-DNA adducts were identified and characterized with liquid-electrospray ionization-quadrupole-time of flight-mass spectrometry (LC-ESI-Q-TOF-MS/MS) and LC-Diode array detector-fluorescence (LC-DAD-FL) analysis. This work provides a robust tool for possible large-scale preparation of AA-DNA adduct standards, which can promote the further studies on carcinogenic and mutagenic mechanism of aristolochic acids. 关 键 词：反相高效液相色谱 DNA加合物 马兜铃酸 固相萃取 制备工艺 串联质谱 二极管阵列检测器 反应混合物
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