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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
May 24 - june 08, 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Performed to guidelines in 2012 with GLP.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report Date:
2012

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Qualifier:
according to
Guideline:
EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)
GLP compliance:
yes (incl. certificate)
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: liquid
Details on test material:
Name: Formaldehyde, reaction product with ethylenediamine
CAS No.: 84066-92-2
Batch No.: PK280-61
Purity: 100%
Storage Conditions: at room temperature

Method

Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Metabolic activation:
with and without
Metabolic activation system:
S9 Mix
Test concentrations with justification for top dose:
The test item was dissolved in A. dest. and diluted prior to treatment. The solvent was compatible with the survival of the bacteria and the S9 activity.
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
sodium azide
methylmethanesulfonate
other: 4-nitro-o-phenylene-diamine; 2-aminoanthracene
Details on test system and experimental conditions:
Five strains of S. typhimurium with the following characteristics were used:
TA 98:
his D 3052; rfa-; uvrB-; R-factor: frame shift mutations
TA 100:
his G 46; rfa-; uvrB-; R-factor: base-pair substitutions
TA 1535:
his G 46; rfa-; uvrB-: base-pair substitutions
TA 1537:
his C 3076; rfa-; uvrB-: frame shift mutations
TA 102:
his G 428 (pAQ1); rfa-; R-factor: base-pair substitutions

Results and discussion

Test results
Key result
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
ambiguous
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

In experiment I toxic effects of the test item were observed in tester strains TA 98, TA 100, TA1535 and TA 1537 at concentrations of 1000 μg/plate and higher (with and without metabolic activation). In tester strain TA 102 toxic effects of the test item were noted at concentrations of 1000 μg/plate and higher (without metabolic activation) and at concentrations of 2500 μg/plate and higher (with metabolic activation). In experiment II toxic effects of the test item were noted in tester strains TA 98 and TA 102 at a concentration of 1500 μg/plate (with and without metabolic activation). In tester strain TA 100 toxic effects of the test item were noted at concentrations of 1000 μg/plate and higher (with and without metabolic activation). In tester strain TA 1535 toxic effects of the test item were observed at concentrations of 750 μg/plate and higher (without metabolic activation) and at concentrations of 250 μg/plate and higher (with metabolic activation). In tester strain TA 1537 toxic effects of the test item were noted at concentrations of 750 μg/plate and higher (with and without metabolic activation).

No biologically relevant increases in revertant colony numbers were noted in tester strains TA 1535, TA 1537 and TA 102. Biologically relevant increases of revertant colony numbers was observed in tester strain TA 100 at a concentration of 316 μg/plate in experiment I (with metabolic activation) but this effect could not be reproduced in experiment II. Biologically relevant increases of revertant colony numbers was also observed in experiment II in tester strain TA 98 at a concentration of 500 μg/plate (with metabolic activation), though this effect was not observed in experiment I. The threshold value of 2.0 was exceeded and a maximum mutation factor of 2.4 was reached at a concentration of 316 μg/plate in experiment I. However, no dose-response relationship was observed for both tester strains. Thus, the observed effects are considered to be equivocal.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
ambiguous with and without metabolic activation

Formaldehyde, reaction product with ethylenediamine
caused gene mutations by frameshifts in the genome of the tester strain TA 98 and by
base pair changes in the genome of the tester strain TA 100, however, in both strains
only in one of two experiments.
A dose-related increase in the number of revertant colonies in tester strains TA 98 and
TA 100 was not observed.
Therefore, due to the observed borderline effects, Formaldehyde, reaction product with
ethylenediamine is considered to be equivocal in this bacterial reverse mutation assay.