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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:
May-July 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: A GLP study conducted under OECD guidelines

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 473 (In Vitro Mammalian Chromosome Aberration Test)
Qualifier:
according to
Guideline:
EU Method B.10 (Mutagenicity - In Vitro Mammalian Chromosome Aberration Test)
Qualifier:
according to
Guideline:
EPA OPPTS 870.5375 - In vitro Mammalian Chromosome Aberration Test
GLP compliance:
yes (incl. certificate)
Type of assay:
in vitro mammalian chromosome aberration test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
Sponsors identification: F-3014
Description: Off white powder
Purity: 100%
Batch number: 290110574
Date received: 10 November 2-11
Storage condition: Room temperature in the dark

Method

Species / strain
Species / strain / cell type:
lymphocytes: human
Details on mammalian cell type (if applicable):
For each experiment, sufficient whole blood was drawn from the peripheral circulation of a volunteer who had been previuosly screened for suitabilityThe volunteer had not been exposed to high levels of radiation or hazardous chemicals and had not knowingly recently suffered from a viral infection. The cell-cycle time for the lymphocytes from the donors used in this study was determined using BrdU (bromodeoxyuridine) incorporation to assess the number of first, second and third division metaphase cells and so circulate the average generation time (AGT). The avarage AGT fot regular donors used in this laboratory has been determined to be approx. 16 hr under typical experimental exposure conditions.



Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
S9 Mix
Test concentrations with justification for top dose:
Preliminary test: dose range used 19.53 to 5000 µg/ml
Experiment 1 (without S-9 mix): 39.06, 78.13, 156.25, 312.5, 625 and 1250 µg/ml. 4 hr exposure to the test item w/o S-9 mix followed by 20 hr culture in treatment free media prior to cell harvest.
Experiment 1 (with S-9 mix): 39.06, 78.13, 156.25, 312.5, 625 and 1250 µg/ml 4 hr exposure to the test item with S-9 mix followed by 20 hr culture in treatment free media prior to cell harvest.
Experiment 2 (without S-9 mix): 39.06, 78.13, 156.25, 312.5, 625 and 1250 µg/ml 24 hr continuous exposure to the test item w/o S9 mix prior to cell harvest.
experiment 2 (with S-9 mix): 39.06, 78.13, 156.25, 312.5, 625 and 1250 µg/ml 4 hr exposure to the test item with S9 mix (2% final conc) followed by 20 hr culture in treatment free media prior to cell harvest.
Vehicle / solvent:
Vehicle(s)/solvent(s) used: DMSO
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
Positive controls:
yes
Remarks:
with and without S9 mix
Positive control substance:
cyclophosphamide
mitomycin C
Details on test system and experimental conditions:
see attached file (test system and conditions)
Statistics:
The frequency of cells with aberrations excluding gaps and the frequency of polyploid cells were compared, were necessary, with the concurrent vehicle control value using Fisher's Exact test.

Results and discussion

Test results
Key result
Species / strain:
lymphocytes: human
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
other: moderate
Vehicle controls validity:
valid
Positive controls validity:
valid
Additional information on results:
All vehicle control groups had frequencies of cells with aberrations within the range expected for normal human lympocytes.
All the positive control items induced statistically significant increases in the frequency of cells with aberrations indicating that the sensitivity of the aassay and the efficacy of the S9 mix were validated.
The test item was moderately non toxic and did not induce any statistically significant increases in the frequency of cells with aberrations, in either of two separate experiments, using a dose range that included a dose level included at lease one precipitating dose level
Remarks on result:
other: strain/cell type: lymphocytes
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

see attached file on results

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative non clastogenic

The test item was moderately non toxic and did not induce any statistically significant increases in the frequency of cells with aberrations, in either of two separate experiments, using a dose range that included a dose level included at lease one precipitating dose level. Therefore the test item F-3014 was considered to be non-clastogenic to human lymphocytes in vitro.
Executive summary:

In vitro study which describes the detection of structural chromosomal aberrations in cultured mammalian cells using OECD 473 and method B10 of the commission regulation.

Duplicate cultures of human lymphocytes, treated with the test item, were evaluated for chromosome aberrations at four dose levels, together with vehicle and positive controls. four treatment conditions were used for the study. With and with out S-9 Mix (2%) (4 hr presence, 20 hr expression period), 4 hr with S-9 mix (1%), 20 hr expression period and exposure time for 24 hr without S-9 mix.

All vehicle control groups had frequencies of cells with aberrations within the range expected for normal human lympocytes.

All the positive control items induced statistically significant increases in the frequency of cells with aberrations indicating that the sensitivity of the aassay and the efficacy of the S9 mix were validated.

The test item was moderately non toxic and did not induce any statistically significant increases in the frequency of cells with aberrations, in either of two separate experiments, using a dose range that included a dose level included at lease one precipitating dose level.

The test item F-3014 was considered to be non-clastogenic to human lymphocytes in vitro.