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A fluorometric-HPLC assay for quantitating the binding of benzo[a]pyrene metabolites to DNA

Authors
Journal
Biochemical and Biophysical Research Communications
0006-291X
Publisher
Elsevier
Publication Date
Volume
109
Issue
1
Identifiers
DOI: 10.1016/0006-291x(82)91594-7
Disciplines
  • Physics

Abstract

Abstract A nonradiometric method is presented for quantitating low levels of benzo[a]pyrene (BP) derivatives that are covalently bound to the DNA of BP-treated mice. This method consists of hydrolyzing the DNA with acid to liberate the BP-adducts in the form of the isomeric tetrols of BP. These tetrols have fluorescence quantum yields of ∼0.7 in deoxygenated solution at 298 K. Hence they are easily quantitated, following HPLC separation, by means of fluorescence detection. The sensitivity of the method is such that one bound BP residue per 10 7 bases can be detected in 100 μg of DNA.

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