Formaldehyde, reaction products with ethylenediamine

Genetic toxicity: in vitro

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 guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

84066-92-2

Cas Number:

84066-92-2

IUPAC Name:

84066-92-2

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.