Measurement of oxidized bases in DNA. Comparison between HPLC-EC and GC-MS assays

被引：13

作者：

Ravanat, JL ^{[1
]}

Douki, T ^{[1
]}

Turesky, R ^{[1
]}

Cadet, J ^{[1
]}

机构：

[1] NESTLE RES CTR,CH-1000 LAUSANNE 26,SWITZERLAND

来源：

JOURNAL DE CHIMIE PHYSIQUE ET DE PHYSICO-CHIMIE BIOLOGIQUE | 1997年 / 94卷 / 02期

关键词：

DNA damage; oxidized DNA bases; GC-MS; HPLC-EC;

D O I：

10.1051/jcp/1997940306

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The detection of oxidized bases in cellular DNA is a challenging problem due to the sensitivity required that should be high enough in order to detect one modification per 10(6) DNA bases. In this respect, gas chromatography mass spectrometry (GC-MS) has been extensively used during the last ten years. However, comparison of the results obtained by this method and those inferred from high performance liquid chromatography analyses coupled with an electrochemical detection (HPLC-EC) indicated an overestimation in the level of the oxidized DNA bases in the GC-MS measurement. A comparative evaluation of the two analytical methods allows us to demonstrate that this overestimation is due to an artefactual oxidation of normal DNA bases during the derivatization reaction that is required prior to GC-MS analysis. Purification of the modified DNA bases prior to the derivatization reaction prevents the artefact to occur. Under these conditions, the results obtained using the GC-MS method and the HPLC-EC assay are similar.

引用

页码：306 / 312

页数：7

共 5 条

[1] Observation and prevention of an artefactual formation of oxidized DNA bases and nucleosides in the GC-EIMS method [J].