2,2-dimethylpropan-1-ol, tribromo derivative; 3-bromo-2,2-bis(bromomethyl)propan-1-ol

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP and appropriate guidelines

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.5395 (In Vivo Mammalian Cytogenetics Tests: Erythrocyte Micronucleus Assay)

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: Commission derective 2000/32/EC, Annex 4C

Principles of method if other than guideline:

not relevant

GLP compliance:

yes (incl. QA statement)

Type of assay:

micronucleus assay

Test material

Test material information

Constituent 1

Reference substance name:

2,2-dimethylpropan-1-ol, tribromo derivative

EC Number:

253-057-0

EC Name:

2,2-dimethylpropan-1-ol, tribromo derivative

Cas Number:

36483-57-5

Molecular formula:

C5H9Br3O

IUPAC Name:

3-bromo-2,2-bis(bromomethyl)propan-1-ol

Details on test material:

Identity: FR-513
chemical name: Tribromoneopentyl Alcohol
batch No.: 39084
Aggregate State at room temperature: Solid
Colour: White to off-white
Mol. weight: 324.84 g/mol
Purity: 98.1%
Solubility: 1.93 g/L at 20°C
Stability: stable under normal conditions
Storage: at room temperature, moisture protected.

Test animals

Species:

mouse

Strain:

NMRI

Sex:

male/female

Details on test animals or test system and environmental conditions:

Source: Harlan Winkelmann GmbH D-33178 Borchen
Number of animals: 81 (45 males/36 females)
Initial age at the start of acclimatisation: 8-9 weeks (males), 11-12 weeks (females)
Acclimatisation: minimum 5 days
initial body weight at start of treatment: Males mean value 32.9g (SD ± 1.9 g); Females mean value 31.3 g (SD ± 2.7 g)
Experiment was conducted under standard lab conditions:
housing: Single
Cage Type: Markon Type I, with wire mesh top
Bedding: granulated soft wood bedding
Feed: pelleted standard diet, ad libitum
Water: tap water, ad libitum
Environment: temperature 22 ± 3° C; Relative humidity 28-74%, artificial light 6 a.m-6 p.m

Administration / exposure

Route of administration:

oral: feed

Vehicle:

DMSO+corn oil (30%-70%)

Details on exposure:

On the day of the experiment, the test item was formulated in DMSO+corn oil (30%-70%). The vehicle was chosen to its relative non-toxicity for the animals. All animals received a single standard volume of 10 mL/kg body weight orally.

Duration of treatment / exposure:

Pre-experiment test: toxic symptoms examined at intervals of aroud 1hr, 2-4hr, 6 hr, 24hr, 30 hr and 48 hr after administration of the test item.
main test: The animals of all dose groups were examined for acute toxic symptoms at intervals of aroud 1 hr, 2-4 hr, 6 and 24 after administration of the test item.

Frequency of treatment:

single administration

Post exposure period:

main test: sampling of bone marrow was done 24 and 48 hr after treatment

Doses / concentrations

Remarks:

Doses / Concentrations:

Basis:
nominal in diet
preliminary test: 2000, 1500, 1000, 500,400, 300 (mg/kg b.w) main test: 300, 150, 75 (mg/kg b.w)

No. of animals per sex per dose:

Ten animals (5 males, 5 females) per dose

Control animals:

yes

Positive control(s):

CPA; Cyclophosphamide (>98%); Dosing: 40 mg/kg b.w ; volume administration: 10 mL/kg b.w

Examinations

Tissues and cell types examined:

Polychromatic erythrocytes (PCEs)

Details of tissue and slide preparation:

Animals were sacrificedusing CO2 following the bleeding. The femora were removed, the epiphyses were cut off and the marrow was flushed out with foetal calf serum, usinga syringe. The cell suspension wass centrifuged at 1500 rpm (390 xg) for 10 minutes and the supernatant was discarded. A small drop of the re-suspended cell pellet was spread on a slide. The smear was air-dried and then stained with May-Grunwald/Giemsa. Cover clips were mounted with EUKITT. At least one slide was made from each bone marrow sample.

Evaluation criteria:

A test item is classified as mutagenic if it induces either a dose related increase or a clear increase in the number of micronucleated polychromatic erythrocytes in a single dose group. Statistical methods (nonparametric Mann-whitney test will be used as an aid in evaluating the results. However, the primary point of consideration is the biological relevance of the results. A test item that fails to produce a biological relevant increase in the number of micronucleated polychromatic erythrocytes is considered non mutagenic in this system.

Statistics:

nonparametric Mann-whitney test will be used as an aid in evaluating the results

Results and discussion

Test results

Test results 1

Sex:

male/female

Genotoxicity:

negative

Remarks:

% micronuclei 0.125 (24 hr)

Toxicity:

yes

Remarks:

(300 mg/kg bw) (1, 2-4, 6, 24, 48 hr post treatment)

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Sex:

male/female

Genotoxicity:

negative

Remarks:

% micronuclei 0.110 (24 hr)

Toxicity:

yes

Remarks:

(150 mg/kg bw) (1, 2-4, 6, 24 hr post treatment)

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Test results 3

Sex:

male/female

Genotoxicity:

negative

Remarks:

% micronuclei 0.085 (24 hr)

Toxicity:

yes

Remarks:

(75 mg/kg bw) ( 2-4 hr post treatment)

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

see attached document on results which includes pre-experiment

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information): negative FR-513 did not induce micronuclei as determined in the test
FR-513 did not induce micronuclei as determined by the micronucleus test with bone marrow cells of the mouse. Therefore FR-513 can be considered to be non - mutagenic in this test.

Executive summary:

see attached document on summary