Deforce D L, Ryniers F P, van den Eeckhout E G, Lemière F, Esmans E L
Laboratory for Pharmaceutical Biotechnology, University of Ghent, Belgium.
Anal Chem. 1996 Oct 15;68(20):3575-84. doi: 10.1021/ac9600013.
The in vitro adduct formation with phenyl glycidyl ethers (PGEs) was studied on 2'-deoxynucleotides and DNA. The modified DNA was hydrolyzed enzymatically, and the mixtures consisting of unmodified 2'-deoxynucleotide adducts were analyzed by capillary zone electrophoresis (CZE), CZE-electrospray mass spectrometry (CZE/ES-MS) and CZE-electrospray tandem mass spectrometry (CZE/ES-MS/MS) using sample stacking. For the CZE analyses, a homemade system was developed in order to enhance the reproducibility of the retention times. This modification enabled the total comparison of the electropherograms obtained for the analysis of 2'deoxynucleotides mixtures with the electropherograms obtained for the DNA hydrolysates both treated with PGEs. The assignment of adducted and nonadducted 2'-deoxynucleotide peaks was unambiguous. Analysis of the CZE/ES-MS data gave the necessary structural information and revealed the presence of mono- and dialkylated 2'-deoxynucleotides. Interpretation of the CZE/ES-MS/MS data of the monoalkylated products allowed differentiation between purine or pyrimidine alkylation and alkylation of the 5-phosphate moiety. Recording of full-scan mass spectra during CZE/ES-MS/MS analysis of 2'-deoxynucleotide reaction mixtures and DNA hydrolysates was possible, using the described CZE sample stacking technique.
研究了苯基缩水甘油醚（PGEs）与2'-脱氧核苷酸和DNA的体外加合物形成情况。对修饰后的DNA进行酶水解，然后使用样品堆积技术，通过毛细管区带电泳（CZE）、CZE-电喷雾质谱（CZE/ES-MS）和CZE-电喷雾串联质谱（CZE/ES-MS/MS）分析由未修饰的2'-脱氧核苷酸加合物组成的混合物。为了提高保留时间的重现性，开发了一种自制系统用于CZE分析。这种改进使得能够对用PGEs处理的2'-脱氧核苷酸混合物分析所得的电泳图与DNA水解产物分析所得的电泳图进行全面比较。加合和未加合的2'-脱氧核苷酸峰的归属明确。对CZE/ES-MS数据的分析提供了必要的结构信息，并揭示了单烷基化和二烷基化2'-脱氧核苷酸的存在。对单烷基化产物的CZE/ES-MS/MS数据的解释使得能够区分嘌呤或嘧啶烷基化与5-磷酸部分的烷基化。使用所述的CZE样品堆积技术，可以在对2'-脱氧核苷酸反应混合物和DNA水解产物进行CZE/ES-MS/MS分析期间记录全扫描质谱。
