Fluorescence HPLC methods for detecting benzo[a]pyrene-7,8-dihydrodiol 9,10-oxide-deoxyadenosine adducts in enzyme-digests of modified DNA

Improved sensitivity

Junxin Chen, Michael C. MacLeod, Ruchen Zhao, Nicholas Geacintov

Research output: Contribution to journalArticle

Abstract

The fluorescence of mononucleoside adducts derived from the binding of anti-7β,8α-dihydroxy-9α,10α-epoxy-7,8,9,10- tetrahydrobenzo[a]pyrene (BPDE I) to N6-deoxyadenosine (BPDE - dA adducts) is 10 -100 times stronger (depending on the methanol/water solvent composition) than the fluorescence of adducts derived from the binding of this diol epoxide derivative to N2-deoxyguanosine. It is shown here that these fluorescence characteristics can be used to quantitate the relatively low yields of BPDE - dA adducts by fluorescence detection when BPDE-modified DNA is subjected to enzymatic degradation to the mononucleoside levels, followed by HPLC analysis of the digests.

Original languageEnglish (US)
Pages (from-to)1049-1051
Number of pages3
JournalCarcinogenesis
Volume14
Issue number5
StatePublished - May 1993

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High-performance Liquid Chromatography
7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide
Pyrene
Fluorescence
Oxides
Enzymes
DNA
High Pressure Liquid Chromatography
Deoxyguanosine
Epoxy
Epoxy Compounds
Methanol
Degradation
Derivatives
Water
Derivative
2'-deoxyadenosine
benzo(a)pyrene 7,8-dihydrodiol
High-performance liquid chromatography
Chemical analysis

ASJC Scopus subject areas

  • Cancer Research
  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Physiology (medical)
  • Physiology
  • Behavioral Neuroscience

Cite this

Fluorescence HPLC methods for detecting benzo[a]pyrene-7,8-dihydrodiol 9,10-oxide-deoxyadenosine adducts in enzyme-digests of modified DNA : Improved sensitivity. / Chen, Junxin; MacLeod, Michael C.; Zhao, Ruchen; Geacintov, Nicholas.

In: Carcinogenesis, Vol. 14, No. 5, 05.1993, p. 1049-1051.

Research output: Contribution to journalArticle

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abstract = "The fluorescence of mononucleoside adducts derived from the binding of anti-7β,8α-dihydroxy-9α,10α-epoxy-7,8,9,10- tetrahydrobenzo[a]pyrene (BPDE I) to N6-deoxyadenosine (BPDE - dA adducts) is 10 -100 times stronger (depending on the methanol/water solvent composition) than the fluorescence of adducts derived from the binding of this diol epoxide derivative to N2-deoxyguanosine. It is shown here that these fluorescence characteristics can be used to quantitate the relatively low yields of BPDE - dA adducts by fluorescence detection when BPDE-modified DNA is subjected to enzymatic degradation to the mononucleoside levels, followed by HPLC analysis of the digests.",
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AB - The fluorescence of mononucleoside adducts derived from the binding of anti-7β,8α-dihydroxy-9α,10α-epoxy-7,8,9,10- tetrahydrobenzo[a]pyrene (BPDE I) to N6-deoxyadenosine (BPDE - dA adducts) is 10 -100 times stronger (depending on the methanol/water solvent composition) than the fluorescence of adducts derived from the binding of this diol epoxide derivative to N2-deoxyguanosine. It is shown here that these fluorescence characteristics can be used to quantitate the relatively low yields of BPDE - dA adducts by fluorescence detection when BPDE-modified DNA is subjected to enzymatic degradation to the mononucleoside levels, followed by HPLC analysis of the digests.

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