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Specific investigations: other studies

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Endpoint:
specific investigations: other studies
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Acceptable publication which meets basic scientific principles

Data source

Referenceopen allclose all

Reference Type:
publication
Title:
32P-Postlabelling of DNA adducts formed by allyl glycidyl ether in vitro and in vivo.
Author:
Plna K and Segerbaeck D
Year:
1997
Bibliographic source:
Carcinogenesis 18(8): 1457-1462.
Reference Type:
publication
Title:
Mini-review specific DNA adducts induced by some mono-substitued epoxides in vito and in vivo.
Author:
Koskinen M and Plna K
Year:
2000
Bibliographic source:
Chemico-biological interactions 129: 209-229.

Materials and methods

Principles of method if other than guideline:
Salmon testes DNA were treated with 500 mM allyl glycidyl ether (AGE) for 24 h at 37°C and analysed for DNA adducts.
Type of method:
in vitro
Endpoint addressed:
genetic toxicity

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Racemic allyl glycidyl ether
- Analytical purity: 99%

Administration / exposure

Duration of treatment / exposure:
24 h
Doses / concentrations
Remarks:
Doses / Concentrations:
500 mM
Basis:

Results and discussion

Details on results:
32P-Postlabelling analysis of AGE-treated DNA after adducts 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.

Any other information on results incl. tables

- AGE-treated DNA was enzymatically digested to deoxyribo- nucleotide-39-monophosphates, enriched by ion-exchange chromatography and 32P-labelled. Two major and one minor adduct spots, absent in untreated DNA, were observed on the autoradiograms.

- HPLC separations of the reaction mixtures revealed that two major products were formed with each nucleotide. Products within each pair were assumed to be diastereomeric, as they were formed in identical yields, had identical UV spectra and merged to one HPLC peak after depurination/depyrimidination.

- The major product with dGMP was that at N-7, with dAMP at N-1 and with dCMP at N-3. The products at N6-adenine and N-3-uracil were prepared by rearrangements from the N-1 adduct of dAMP and the N-3 adduct of dCMP, respectively.

- 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 AGE-modified DNA was 9–12%. The labeling efficiency of the 1-adenine adduct standard was 78–82%.

- Total recovery of this adduct from DNA was ~20% when using anion-exchange chromatography for adduct enrichment and 30–34% when using nuclease P1.

Applicant's summary and conclusion