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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro cytogenicity / chromosome aberration study in mammalian cells
Remarks:
Type of genotoxicity: chromosome aberration
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study and GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
Qualifier:
according to
Guideline:
EU Method B.10 (Mutagenicity - In Vitro Mammalian Chromosome Aberration Test)
GLP compliance:
yes
Type of assay:
in vitro mammalian chromosome aberration test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
CAS: 7085-85-0
Molecula weight: 125.13
Description: clear colorless liquid
Batch: L2PF090240
Purity: 99.7%

Method

Target gene:
no data
Species / strain
Species / strain / cell type:
human lymphoblastoid cells (TK6)
Metabolic activation:
with and without
Metabolic activation system:
S9 mix
Test concentrations with justification for top dose:
320, 640 and 1280 µg/ml culture medium
Vehicle / solvent:
- Vehicle/solvent used: acetone
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: Mitomycin C without S9, Cyclophosphamide with S9
Details on test system and experimental conditions:
Test system:
Cultured peripheral human lymphocytes were used as test system. Blood was collected from healthy adult, non-smoking, male volunteers. The Average Generation time (AGT) of the cells and the age of the donor at the time the AGt was determined (December 2009) are presented below:
- First dose range finding study: age 27, AGT = 14.3 h
- Second range finding study: age 33, AGT = 14.4 h
- First cytogenetic assay: age 33, AGT = 14.4 h
- Second cytogenetic assay: age 39, AGT = 14.0 h

All incubations were carried out in a controlled envrionment in the dark, in which optimal conditions were a humid atmosphere of 80-100%, containing 5.0 +/- 0.5% CO2 in air, at a temperature of 37.0 +/- 1.0°C.

Results and discussion

Test results
Species / strain:
human lymphoblastoid cells (TK6)
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
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

No effects of ethyl-2-cyanoacrylate on the number of polyploid cells and cells with endoreduplicated chromosomes were observed both i n the absence and presence of S9 -mix. Therefore it can be concluded that ethyl-2-cyanoacrylate does not disturb mitotic processes and cell cycle progression and does not induce numerical chromosome aberrations under the experimental conditions described in this report.
Executive summary:

This study was performed to investigate the potential of ethyl-2-cyanoacrylate to induce chromosome aberrations in cultured peripheral human lymphocytes. The study were based on the most recent OECD and EC guidelines.

This report describes the effect of ethyl-2-cyanoacrylate on the number of chromosome aberrations in cultured peripheral human lymphocytes in the presence and absence of a metabolic activation system (phenobarbital and naphthoflavone induced rat liver S9 -mix). The possible clastogenicity of ethyl-2-cyanoacrylate was tested in two independent experiments.

A stock solution of 512 mg/ml and 611 mg/ml ethyl-2-cyanoacrylate was prepared in acetone for the use in the absence and presence of S9 -mix, respectively. Subsequent dilutions in culture medium resulted in the formation of precipiates.

In the first cytogentic assay, ethyl-2-cyanoacrylate was tested up to a loading rate of 1280 µg/ml (ca.0.01 M) for a 3 h exposure time with a 24 h fixation time in the absence and presence of 1.8% (v/v) S9 -fraction.

In the second cytogenetic assy, ethyl-2-cyanoacrylate was again tested up to a loading rate of 1280 µg/ml for a 24 h and 48 h continuous exposure timewith a 24 h and 48 fixation time in the absence of S9 mix. in the presence of S9 mix ethyl-2-cyanoacrylate was tested at the same concnetration range for a 3 h exposure time with a 48 h fixation time.

The number of cells with chromosome aberrations found in the negative, solvent and hydroquinone control cultures was within the laboratory historical control data range. Positive control chemicals, mitomycin C and cyclophosphamide, both produced a statistically significant increase in the incidence of cells with chromosome aberrations, indicating that the test conditions were adequate and that the metabolic activation system (S9 -mix) functioned properly.

ethyl-2-cyanoacrylate did not induce a statistically significant or biologically relevant increase in the number of cells with chromosome aberrations in the absence and presence of S9 -mix, in either of the two independently repeated experiments.

No effects of ethyl-2-cyanoacrylate on the number of polyploid cells and cells with endoreduplicated chromosomes were observed both i n the absence and presence of S9 -mix. Therefore it can be concluded that ethyl-2-cyanoacrylate does not disturb mitotic processes and cell cycle progression and does not induce numerical chromosome aberrations under the experimental conditions described in this report.

Finally, it is concluded that this test is valid and that ethyl-2-cyanoacrylate is not clastogenic in human lymphocytes under experimental conditions described in this report.