Back to Search
Start Over
Detection of the acrolein-derived cyclic DNA adduct by a quantitative 32P-postlabeling/solid-phase extraction/HPLC method: Blocking its artifact formation with glutathione
- Source :
-
Analytical Biochemistry . Mar2008, Vol. 374 Issue 1, p163-172. 10p. - Publication Year :
- 2008
-
Abstract
- Abstract: Acrolein (Acr), a hazardous air pollutant, reacts readily with deoxyguanosine (dG) in DNA to produce cyclic 1, N 2-propanodeoxyguanosine adducts (Acr-dG). Studies demonstrate that these adducts are detected in vivo and may play a role in mutagenesis and carcinogenesis. In the study described here, a quantitative 32P-postlabeling/solid-phase extraction/HPLC method was developed by optimizing the solid-phase extraction and the 32P-postlabeling conditions for analysis of Acr-dG in DNA samples with a detection limit of 0.1fmol. It was found that Acr-dG can form as an artifact during the assay. Evidence obtained from mass spectrometry indicates that the Acr in water used in the assay is a likely source of artifact formation of Acr-dG. The formation of Acr-dG as an artifact can be effectively blocked by adding glutathione (GSH) to the DNA sample to be analyzed. In addition, Acr-dG was detected as a contaminant in the commercial dG and dT 3′-monophosphate samples. Finally, this method was used to detect Acr-dG in calf thymus and human colon HT29 cell DNA with an excellent linear quantitative relationship. [Copyright &y& Elsevier]
- Subjects :
- *ACROLEIN
*ALDEHYDES
*HERBICIDES
*DNA
Subjects
Details
- Language :
- English
- ISSN :
- 00032697
- Volume :
- 374
- Issue :
- 1
- Database :
- Academic Search Index
- Journal :
- Analytical Biochemistry
- Publication Type :
- Academic Journal
- Accession number :
- 29370430
- Full Text :
- https://doi.org/10.1016/j.ab.2007.10.029