TITLE

Styrene oxide DNA adducts: quantitative determination using31P monitoring

AUTHOR(S)
Edler, Michael; Jakubowski, Norbert; Linscheid, Michael
PUB. DATE
January 2005
SOURCE
Analytical & Bioanalytical Chemistry;Jan2005, Vol. 381 Issue 1, p205
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
ABSTRACT
The reaction of styrene oxide, a potential carcinogen in humans, with DNA constituents has been used to develop an improved method for quantification of DNA adducts. To enable monitoring of DNA adducts caused by xenobiotics at physiological relevant levels, a robust, reliable and powerful method based on monitoring of phosphorus in nucleotides is described. An efficient enzymatic digestion step and a sample-preconcentration procedure are essential, and enable separation of alkylated nucleotides from the large excess of native nucleotides. The adducts are detected by means of the phosphorus signal measured at massm/z=31 with an inductively-coupled-plasma mass spectrometer. Bis(4-nitrophenyl)phosphate (BNPP) serves as internal standard for quantification of the adducts. The absolute limit of detection, 45 fmol, corresponds to detection of three modified nucleotides among 107 native nucleotides (the calculation is based on use of 50 µg calf thymus DNA). An adduct formation ratio at the DNA of 3.6 adducts per 1000 nucleotides was measured, which is 75% lower than for reaction with monomeric 2'-deoxy-nucleotides. In addition, a substantial amount of phosphate adducts were detected, but in DNA the rate of phosphate formation was lower than with monomeric nucleotides. Most probably these adducts escaped unnoticed when31P-post-labelling was employed.
ACCESSION #
16004434

 

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