Carcinogenesis, Vol 18, 1457-1462, Copyright © 1997 by Oxford University Press
K Plna and D Segerback
32P-Postlabelling analysis of allyl glycidyl ether-treated DNA after adduct
enrichment on anion-exchange cartridges revealed two major and one minor
DNA adducts. The major adducts were shown to originate from alkylation at
N-7-guanine and N-1-adenine, respectively, while the minor adduct was at
N-3-cytosine. In addition, rearrangement products of the 1-adenine and
3-cytosine adducts to N6-adenine and 3-uracil were indicated. The relative
amounts of adenine, cytosine and uracil products appeared to be dependent
upon conditions (in particular pH) during sample processing and analysis.
When nuclease P1 was used for adduct enrichment the adenine, cytosine and
uracil adducts, but not the 7-guanine adduct, were detected. The labelling
efficiency of the 7- guanine adduct standard was 40-45%. Total recovery of
this adduct from allyl glycidyl ether-modified DNA was 9-12%. The labelling
efficiency of the 1-adenine adduct standard was 78-82%. Total recovery of
this adduct from DNA was approximately 20% when using anion-exchange
chromatography for adduct enrichment and 30-34% when using nuclease P1.
Preliminary analysis of DNA from mice treated with allyl glycidyl ether
indicated 57 times higher level of the 7-guanine adduct, per unit dose, in
skin DNA (120 per 10(8) normal nucleotides) after topical application when
compared to liver DNA after i.p. administration. The 1- adenine adduct
could not be quantified in liver DNA (due to an interfering background
product present in untreated animals) and the level of the 3-cytosine
adduct was below the detection limit of the method. After topical
application the level of the 1 adenine adduct in skin DNA was approximately
30 per 10(8), using either column or nuclease P1 enrichment. The 3-cytosine
adduct was detected in skin, but was not quantified.
ARTICLES
32P-Postlabelling of DNA adducts formed by allyl glycidyl ether in vitro and in vivo
Center for Nutrition and Toxicology, Department of Biosciences, Karolinska Institute, Novum, Huddinge, Sweden.
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