Quantitative measurement of radiation-induced base products in DNA using gas chromatography-mass spectrometry

Technical Report · Sun Jan 01 04:00:00 EST 1989

OSTI ID:7018563

Fuciarelli, A F; Wegher, B J; Gajewski, E; Dizdaroglu, M; Blakely, W F

Gas chromatography-mass spectrometry with selected-ion monitoring was used to study radiation-induced damage to DNA. Quantitative analysis of modified purine and pyrimidine bases resulting from exposure to ionizing radiation using this technique is dependent upon the selection of appropriate internal standards and calibration of the mass spectrometer for its response to known quantities of the internal standards and the products of interest. The compounds 6-azathymine and 8-azaadenine were found to be suitable internal standards for quantitative measurements of base damage in DNA. For the purpose of calibration of the mass spectrometer, relative molar response factors for intense characteristics ions were determined for the trimethylsilyl derivatives of 5-hydroxyuracil, thymine glycol, and 5,6 dihydrothymine using 6-azathymine, and for the trimethylsilyl derivatives of 4,6-diamino-5-formamidopyrimidine, 8-hydroxyguanine using 8-azaadenine. Accurate measurements of the yield of radiation-induced modifications to the DNA bases is also dependent upon two chemical steps in which the purines and pyrimidines are released from the sugar-phosphate backbone and the derivatized to make them volatile for gas chromatography. The completeness of these reactions, in addition to assessing the stability of the modified DNA bases in acid and their trimethylsilylated derivatives over the time necessary to complete the experimental analysis, was also examined.

• Research Organization: Armed Forces Radiobiology Research Inst., Bethesda, MD (USA)
• OSTI ID: 7018563
• Report Number(s): AD-A-214233/9/XAB; AFRRI-SR--89-30
• Country of Publication: United States
• Language: English

Related Subjects

560120* -- Radiation Effects on Biochemicals
Cells
& Tissue Culture
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ACCURACY
ALKYL RADICALS
AMINES
AROMATICS
AZAARENES
AZINES
BIOLOGICAL EFFECTS
BIOLOGICAL RADIATION EFFECTS
CALIBRATION
CHARGED PARTICLES
CHEMICAL ANALYSIS
CHEMICAL BONDS
CHROMATOGRAPHY
DNA
ELEMENTS
GUANINE
HETEROCYCLIC COMPOUNDS
HYDROXY COMPOUNDS
IONIZING RADIATIONS
IONS
MASS SPECTROSCOPY
METHYL RADICALS
MODIFICATIONS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PURINES
PYRIMIDINES
QUANTITATIVE CHEMICAL ANALYSIS
RADIATION EFFECTS
RADIATIONS
RADICALS
SEMIMETALS
SEPARATION PROCESSES
SILICON
SPECTROSCOPY
STANDARDS
URACILS