1,4-divinylbenzene

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

Data is from peer reviewed publication

Data source

Reference

Reference Type:

publication

Title:

Gene mutation toxicity study of the the test chemical

Author:

McMohan et al

Year:

1979

Bibliographic source:

Cancer Research

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

Modified method

Principles of method if other than guideline:

Gene mutation toxicity study was performed to determine the mutagenic nature of the test chemical

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

1-bromo-4-methoxybenzene

Cas Number:

104-92-7

Molecular formula:

C7H7BrO

IUPAC Name:

1-bromo-4-methoxybenzene

Details on test material:

- Name of test material: p-bromoanisole- Molecular formula: C7H7BrO- Molecular weight: 187.035 g/mol- Substance type: Inorganic- Physical state: No data- Purity: No data- Impurities (identity and concentrations): No data

Method

Target gene:

Histidine for Salmonella typhimurium strains and tryptophan for E. coli strains

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium, other: G46, TA1535, TA100, C3076, TA1537, D3052, TA1538, TA98

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

other:

Species / strain 2

Species / strain / cell type:

E. coli, other: WP2 and WP2 uvrA-

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

other: WP2 (E. coli tryptophan) and WP2 uvrA (E. coli tryptophan with uvrA deletion).

Cytokinesis block (if used):

No data

Metabolic activation:

with and without

Metabolic activation system:

Liver enzymes were prepared from adult male Fischer rats predosed with Aroclor 1254

Test concentrations with justification for top dose:

0.1 – 1000 µg/mL

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO, water or dimethoxyethane- Justification for choice of solvent/vehicle: Solubility

Controls

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

Details not specified

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

2-acetylaminofluorene

other: Streptozotocin (without microsomal activation)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (gradient plate method)DURATION- Preincubation period: No data- Exposure duration: 48 hrs- Expression time (cells in growth medium): 48 hrs- Selection time (if incubation with a selection agent): No data- Fixation time (start of exposure up to fixation or harvest of cells): No dataSELECTION AGENT (mutation assays): No dataSPINDLE INHIBITOR (cytogenetic assays): No dataSTAIN (for cytogenetic assays): No dataNUMBER OF REPLICATIONS: Duplicate sets of plates were prepared: one set in which the test compound was in the bottom wedge and had to diffuse upward and one in which it was in the upper wedge and had to diffuse downward.NUMBER OF CELLS EVALUATED: No dataDETERMINATION OF CYTOTOXICITY- Method: mitotic index; cloning efficiency; relative total growth; other: No dataOTHER EXAMINATIONS:- Determination of polyploidy: No data- Determination of endoreplication: No data- Other: No dataOTHER: No data

Rationale for test conditions:

No data

Evaluation criteria:

Bacterial growth was observed along the streak line and the concentration range over which chemically induced mutant colonies are present is recorded.

Statistics:

No data

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium, other: G46, TA1535, TA100, C3076, TA1537, D3052, TA1538, TA98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Test results 2

Species / strain:

E. coli, other: WP2 and WP2 uvrA-

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

No data

Remarks on result:

other: No mutagenic potential

Applicant's summary and conclusion

Conclusions:

The test chemical did not induce gene mutation in Salmonella typhimurium G46, TA1535, TA100, C3076, TA1537, D3052, TA1538, TA98 and E. coli WP2 and WP2 uvrA- in the presence and absence of Liver enzymes activation system and hence the chemical is not likely to classify as a gene mutant in vitro.

Executive summary:

Gene mutation toxicity study was performed to determine the mutagenic nature of the test chemical. The study was performed using Salmonella typhimurium G46, TA1535, TA100, C3076, TA1537, D3052, TA1538, TA98 and E. coli WP2 and WP2 uvrA- with and without Liver enzymes activation system.

 

Ten ml of minimal agar medium (not containing test compound) was poured into a square Petri dish (9 x 9 cm) which is tilted at a slight angle. The agar was then allowed to solidify into a wedge-shaped layer by standing at room temperature. Meanwhile, a 1000-µg/ml mixture of test compound in agar was prepared by adding 10 ml of minimal agar to 0.1 ml of a 100-mg/mI solution of test compound in dimethyl sulfoxide. When appropriate, water or dimethoxyethane was used instead of dimethyl sulfoxide. The cooled agar plates were then placed on a level surface, and an overlay of the 10 ml of agar containing the test compound was poured onto the plate to form a reversed wedge of agar on top of the first wedge. A concentration gradient of compound was produced by allowing the compound in the upper wedge to diffuse into the lower layer for 2 hr at room temperature. The concentration range in this plate is approximately 100 to 1000µg/ml. Three additional plates with concentration ranges of 10 to 100µg/ml, 1 to 10µg/ml, and 0.1 to 1µg/ml were prepared.

 

A streaking device consisting of 10 sterile 50-µL pipets was dipped into suspensions of the 10 test strains and allowed to fill by capillary action. The pipets were then touched to the upper edge of the gradient and drawn across the plate. The study was performed in the presence and absence of liver enzyle activating system and the plates were incubated for 48 hrs at 37°C.

 

The test chemical did not induce gene mutation in Salmonella typhimurium G46, TA1535, TA100, C3076, TA1537, D3052, TA1538, TA98 and E. coli WP2 and WP2 uvrA- in the presence and absence of Liver enzymes activation system and hence the chemical is not likely to classify as a gene mutant in vitro.