N-(1,1,3,3-tetramethylbutyl)acrylamide

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:

1998-03-18 to 1998-04-13

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study in accordance with OECD TG 409

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1998

Report date:

1998

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Principles of method if other than guideline:

The experimental materials, methods and procedures were based on those described by Ames et al. (1975) and Green and Muriel (1976)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

N-(1,1,3,3-tetramethylbutyl)acrylamide

EC Number:

224-169-7

EC Name:

N-(1,1,3,3-tetramethylbutyl)acrylamide

Cas Number:

4223-03-4

Molecular formula:

C11H21NO

IUPAC Name:

N-(1,1,3,3-tetramethylbutyl)acrylamide

Details on test material:

- Name of test material (as cited in study report): N-tert-Octylacrylamide (937-64)
- Physical state: off-white, fluffy crystalline solid
- Analytical purity: not reported
- Impurities (identity and concentrations): not reported
- Purity test date: not reported
- Lot/batch No.: not reported
- Expiration date of the lot/batch: not reported
- Date received: 13 March 1998

Method

Target gene:

Histidine locus in Salmonella typhimurium and tryptophan locus in Escherichia coli

Species / strain

Species / strain / cell type:

other: S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2 uvrA

Additional strain / cell type characteristics:

other: S. typhimurium strains containalso the rfa wall mutation and a deletion of the uvrB gene; strains TA 89 and TA100 also contain the R-factor plasmid, pKM101; E. coli strain contains a uvrA DNA repair deficiency

Metabolic activation:

with and without

Metabolic activation system:

S9 mix

Test concentrations with justification for top dose:

33.3, 100, 333, 1000, 3330 and 5000 µg per plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: not justified, but vehicle is commonly used in Ames tests

Controls

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

other: With S9 mix: benzyo(a)pyrene (TA98), 2-aminoanthracene (TA100, TA1535, TA1537, WP2uvrA); without S9 mix: 2-nitrofluorene (TA98), sodium azide (TA100, TA1535), ICR-191 (TA1537), 4-nitroquinoline-N-oxide (WP2uvrA)

Remarks:

Other: sterility controls on test article and S9 mix

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 52 ± 4 hours

NUMBER OF REPLICATIONS: 3

DETERMINATION OF CYTOTOXICITY
- Method: Observation of a reduction of revertants per plate and/or a thinning or disappearance of the bacterial background lawn

Evaluation criteria:

Tester strains TA98, TA100, WP2uvrA: Test substance has to produce at least a 2-fold increase in the mean number of revertants per plate of at least one of the these tester strains over the mean number of revertants per plate of the vehicle control. In addition, a dose-response relationship to increasing concentrations of the test substance has to be observed.

Tester strains TA1535, TA1537: Test substance has to produce at least a 3-fold incease in the mean number of revertants per plate of at least one of these tester strains over the mean number of revertants pr plate of the vehicle control. In addition, a dose-response relationship to increasing concentrations of the test substance has to be observed.

Results and discussion

Test results

Species / strain:

other: S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2 uvrA

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 5000 µg per plate in TA100; no cytotoxicity in WP2uvrA

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

All criteria for valid study were fulfilled.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

The results of the Bacterial Reverse Mutation Assay indicate that, under the conditions of this study, N-tert-octylacrylamide did not cause a positive response in the presence and absence of Aroclor-induced rat liver S9 mix.

Executive summary:

The in-vitro potential of N-tert-Octylacrylamide for mutagenic activity in the Salmonella - Escherichia coli/Mammalian-Microsome Reverse Mutation Assay with a Confirmatory Assay was studied under GLP according to OECD TG 471. This assay evaluated the test article and/or its metabolites for their ability to induce reverse mutations at the histidine locus in the genome of specific S. typhimurium tester strains and at the tryptophan locus in an Escherichia coli tester strain both in the presence and absence of an exogenous metabolic activation system of mammalian microsomal enzymes derived from Aroclor-induced rat liver (S9 mix). The doses tested in the mutagenicity assay were selected based on the results of a dose rangefinding study using tester strains TA100 and WP2uvrA and ten doses of test article ranging from 5,000 to 6.67 µg per plate, one plate per dose, both in the presence and absence of S9 mix.

The tester strains used in the mutagenicity assay were Salmonella typhimurium tester strains TA98, TA100, TA 1535, TA1537, and Escherichia coli tester strain WP2uvrA. The assay was conducted with six doses of test article in both the presence and absence of S9 mix along with concurrent vehicle and positive controls using three plates per dose. The doses tested were 5000, 3330, 1000, 333, 100 and 33.3 µg per plate in both the presence and absence of S9 mix. The results of the initial mutagenicity assay were confirmed in an independent experiment. The results of the Salmonella - Escherichia coli/Mammalian-Microsome Reverse Mutation Assay with a Confirmatory Assay indicate that under the conditions of this study, N-tert-Octylacrylamide, did not cause a positive increase in the number of revertants per plate of any of the tester strains either in the presence or absence of microsomal enzymes prepared from Aroclor-induced rat liver (S9).