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激光诱导荧光毛细管电泳在分析dGMP - BPDE加合物中的应用。

Applications of capillary electrophoresis with laser-induced fluorescence for analysis of dGMP-BPDE adduct.

作者信息

Formenton-Catai Ana Paula, Carrilho Emanuel

机构信息

Instituto de Química de São Carlos, Universidade de São Paulo, Brazil.

出版信息

Anal Bioanal Chem. 2003 May;376(1):138-41. doi: 10.1007/s00216-003-1875-4. Epub 2003 Mar 29.

Abstract

DNA adducts are thought to be crucial to the initiation of mutational and carcinogenic processes. Polycyclic aromatic hydrocarbons (PAHs) have been identified as one major source of carcinogenic risk since they can bind to DNA thus forming an adduct. Quantification of this adduct is important because it may correlate to the risk for cancer development. In this study, the adduct formed between 2'-deoxyguanosine 5'-monophosphate and benzo[ a]pyrene-7,8-dihydrodiol-9,10-epoxide (BPDE) was analyzed by capillary electrophoresis. Both capillary zone electrophoresis (CZE) and micellar electrokinetic capillary chromatography (MECC) modes with laser-induced fluorescence detection were used for the separation and analysis of DNA adducts. The exploration of capillary electrophoresis in several modes provided different separation mechanisms in which the stereochemical forms of the adduct could be separated. The best result obtained was using a coated fused-silica capillary in Tris-TAPS buffer, which provided high sensitivity with a detection limit of 2.5x10(-9) mol L(-1). MECC separation of the BPDE adduct, although less sensitive, provided an efficient enantioselective separation option.

摘要

DNA加合物被认为对突变和致癌过程的起始至关重要。多环芳烃(PAHs)已被确定为致癌风险的一个主要来源,因为它们可以与DNA结合从而形成加合物。对这种加合物进行定量很重要,因为它可能与癌症发生风险相关。在本研究中,通过毛细管电泳分析了5'-单磷酸-2'-脱氧鸟苷与苯并[a]芘-7,8-二氢二醇-9,10-环氧化物(BPDE)之间形成的加合物。采用毛细管区带电泳(CZE)和胶束电动毛细管色谱(MECC)模式并结合激光诱导荧光检测对DNA加合物进行分离和分析。对几种模式下毛细管电泳的探索提供了不同的分离机制,借此可以分离加合物的立体化学形式。获得的最佳结果是在Tris-TAPS缓冲液中使用涂覆的熔融石英毛细管,其检测限为2.5×10⁻⁹ mol L⁻¹,具有高灵敏度。BPDE加合物的MECC分离虽然灵敏度较低,但提供了一种有效的对映体选择性分离方法。

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