Determination of pyrimidine dimers in DNA by high-performance liquid chromatography/gas chromatography and electron capture detection
- Oak Ridge National Lab., TN (USA)
Exposure of DNA to uv radiation results in the formation of a number of photoproducts including the cyclobutyl pyrimidine dimers. At low uv fluences the concentrations of these dimeric compounds are only a small fraction of the corresponding DNA pyrimidine concentration (e.g., as low as 0.02% or less of the total thymine content). Sensitive methods of analysis are therefore required for accurate determinations. Analytical methodology based upon HPLC fractionation and electrophore labeling followed by GC/electron capture detection (ECD) has been developed to quantitate these species. Separation of thymine-thymine, thymine-uracil, and uracil-uracil from the monomeric bases and from other constituents present in acid-hydrolyzed DNA is achieved by reversed-phase HPLC. Isolation of the dimeric fractions is followed by off-line derivatization to form pentafluorobenzyl products for analysis by GC/ECD. All active hydrogens are alkylated, yielding products with high response factors and detection limits in the low femtomole range. The overall analytical scheme for the determination of pyrimidine dimers in DNA is presented.
- OSTI ID:
- 7244391
- Journal Information:
- Analytical Biochemistry; (USA), Journal Name: Analytical Biochemistry; (USA) Vol. 182:2; ISSN ANBCA; ISSN 0003-2697
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
Cells
& Tissue Culture
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
CAPTURE
CHEMICAL RADIATION EFFECTS
CHEMISTRY
CHROMATOGRAPHY
DNA
ELECTROMAGNETIC RADIATION
ELECTRON CAPTURE
GAS CHROMATOGRAPHY
LIQUID COLUMN CHROMATOGRAPHY
MEASURING METHODS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PYRIMIDINE DIMERS
RADIATION CHEMISTRY
RADIATION EFFECTS
RADIATIONS
SEPARATION PROCESSES
ULTRAVIOLET RADIATION