2-tert-butylaminoethyl methacrylate

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in mammalian cells

Remarks:

Type of genotoxicity: gene mutation

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study.

Data source

Reference

Reference 1

Reference Type:

study report

Title:

Unnamed

Year:

1992

Report date:

1992

Reference 2

Reference Type:

secondary source

Title:

Atochem (1992), HPRT Gene Mutation Assay in CHO CeIIs, CIT 8515 MVA

Author:

Atochem

Year:

2003

Bibliographic source:

cited in: OECD SIDS, 2-Dimethylaminoethylmethacrylate, CAS No: 2867-47-2, 07/2003

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)

GLP compliance:

yes

Type of assay:

mammalian cell gene mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2-dimethylaminoethyl methacrylate

EC Number:

220-688-8

EC Name:

2-dimethylaminoethyl methacrylate

Cas Number:

2867-47-2

Molecular formula:

C8H15NO2

IUPAC Name:

2-(dimethylamino)ethyl methacrylate

Details on test material:

- Name of test material (as cited in study report): N,N-Dimethylaminoethyl methacrylate (MADAME)
- Supplier: Atochem
- Purity: 99.67%
- Physical state: colourless liquid
- Lot/batch No.: RN 132 - 13/11/91
- Expiration date of the lot/batch: December 1992
- Storage condition of test material: +4°C in the dark

Method

Target gene:

HGPRT

Species / strain

Species / strain / cell type:

Chinese hamster lung fibroblasts (V79)

Metabolic activation:

with and without

Metabolic activation system:

S9 mix: microsomal rat liver portion and cofactors

Test concentrations with justification for top dose:

With S9 mix: 62.5, 125, 250, 500, 1000, 1500, 2000 µg/mL
Without S9 mix: 31.25, 62.5, 125, 250, 500 µg/mL

Vehicle / solvent:

DMSO

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Remarks:

Without S-9: 2.5 µg/ml MNNG. With S-9: 4 µg/ml BaP.

Details on test system and experimental conditions:

Duplicate cultures were used for each experimental point. The cells were seeded at approximately 5 x 10E+5/25 cm² and placed in an incubator at 37°C. Tw enty-four hours later, they were exposed for 3 hours to the TS, either in a medium without fetal calf serum (assay without S9 mix) or in the metabolic activation system (assay with S9 mix). After treatment, the cultures were observed under a microscope for any morphological alterations, the medium was removed and the cells were rinsed with PBS. Then the cells were used for cytotoxicity (cloning efficiency) and mutagenicity tests.

Evaluation criteria:

A test substance is considered as non-mutagenic if it does not induce a mutation frequency that is at least 3-times higher than the mutation frequency of the negative and/or solvent controls.
A test substance is considered as mutagenic if it induces a 3-fold increase in the mutation frequency when compared to the mutation frequency of the negative and/or solvent controls. In this case, a dose relationship is investigated and considered as significant if p < 0.05.
The results are considered as ambigous if a large difference is obtained between the two tests.

Results and discussion

Test results

Species / strain:

Chinese hamster lung fibroblasts (V79)

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

≥ 1000 µg/mL, round and refringent cells from 200 µg/mL on.

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

Although round and refringent cells were observed at 250 µg/mL, the mutation frequency in the cells from duplicate cultures treated with the TS was considered as similar to that of the negative and solvent controls, with and without S9, i.e. no significant increase (3-fold increase over the controls) was observed. The TS did not show mutagenic activity in this HPRT gene mutation assay in V79 Chinese hamster cells.

ADDITIONAL INFORMATION ON CYTOTOXICITY:
By the preliminary cytotoxicity test, the cytotoxicity (decrease in the cloning efficiency and/or dead cells) was shown at the concentrations of equal or greater than 1000 µg/mL, both with or without S9 mix. At 250 µg/mL or higher, round and refringent cells were observed.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

Under the conditions of this study the TS did not show mutagenic activity in the HPRT gene mutation assay in V79 Chinese hamster cells.

Executive summary:

The study was performed according to OECD TG 476 in compliance with GLP.

The test was conducted at concentrations of 31.25 to 2000 µg/mL. With and without metabolic activation, the TS showed some cytotoxic effects at concentrations higher than 250 µg/mL, but no inerease in the mutation frequencies were observed at any concentrations tested.

Conclusion: Under the conditions of this study the TS did not show mutagenic activity in the HPRT gene mutation assay in V79 Chinese hamster cells.