2,2',6,6'-Tetrabromo-4,4'-isopropylidenediphenol, oligomeric reaction products with 1-chloro-2,3-epoxypropane and 2,4,6-tribromophenol

Genetic toxicity: in vitro

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 guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.10 (Mutagenicity - In Vitro Mammalian Chromosome Aberration Test)

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.5375 - In vitro Mammalian Chromosome Aberration Test

GLP compliance:

yes (incl. QA statement)

Type of assay:

in vitro mammalian chromosome aberration test

Test material

Test material information

Constituent 1

Reference substance name:

2,2',6,6'-Tetrabromo-4,4'-isopropylidenediphenol, oligomeric reaction products with 1-chloro-2,3-epoxypropane and 2,4,6-tribromophenol

EC Number:

500-399-6

EC Name:

2,2',6,6'-Tetrabromo-4,4'-isopropylidenediphenol, oligomeric reaction products with 1-chloro-2,3-epoxypropane and 2,4,6-tribromophenol

Cas Number:

158725-44-1

IUPAC Name:

NA

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.