Quantitative analysis of low levels of benzo(a)pyrene diol epoxide bound to DNA: acid-induced liberation of tetraols followed by HPLC and fluorometric detection
Under normal environmental conditions, outside the laboratory, neither the DNA of an organism nor the carcinogens to which it may be exposed are radioactively labeled; hence, the conventional array of radiometric methods for analyzing DNA damage are not applicable. Because of the intrinsic fluorescence associated with B and its metabolites. The use of fluorescence detection for quantitating the amount of BPDE bound to the DNA of exposed animals was explored. Such techniques may have application to the detection of other fluorescent carcinogens as well. Assuming that 100 ..mu..g of DNA can be isolated from the back of a single treated mouse, then a detection limit of 5 pg of tetraol represents a binding level of 5 molecules of BP per 10/sup 8/ bases. This level of sensitivity is about a factor of 10 less than that calculated by Baird for 100 ..mu..g of DNA employing a specific activity of 20 Ci per mmol tritiated BP. Additional improvements in the fluorescence detection system are planned, which could increase the sensitivity by another order of magnitude.
- Research Organization:
- Oak Ridge National Lab., TN (USA)
- DOE Contract Number:
- W-7405-ENG-26
- OSTI ID:
- 5293768
- Report Number(s):
- CONF-820439-4; ON: DE82017437
- Resource Relation:
- Conference: Symposium on genetics mechanisms of carcinogenesis, Gatlinburg, TN, USA, 11 Apr 1982
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
37 INORGANIC
ORGANIC
PHYSICAL AND ANALYTICAL CHEMISTRY
BENZOPYRENE
FLUORESCENCE SPECTROSCOPY
LIQUID COLUMN CHROMATOGRAPHY
SENSITIVITY
BINDING ENERGY
DNA
EPOXIDES
GLYCOLS
LABORATORY ANIMALS
METABOLITES
QUANTITATIVE CHEMICAL ANALYSIS
ALCOHOLS
AROMATICS
CHEMICAL ANALYSIS
CHROMATOGRAPHY
CONDENSED AROMATICS
EMISSION SPECTROSCOPY
ENERGY
HYDROCARBONS
HYDROXY COMPOUNDS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
ORGANIC OXYGEN COMPOUNDS
SEPARATION PROCESSES
SPECTROSCOPY
560305* - Chemicals Metabolism & Toxicology- Vertebrates- (-1987)
400104 - Spectral Procedures- (-1987)