Divinylbenzene

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Data is from J-check

Data source

Reference

Reference Type:

review article or handbook

Title:

In Vitro Chromosomal Aberration Test of Divinylbenzene on Cultured Chinese Hamster C ells

Author:

J-check

Year:

2003

Bibliographic source:

Ministry of Health and Welfare, Japan, 2003

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

Guidelines for Screening Toxicity Testings of Chemicals (Japan) and OECD Guidelines No. 471 and 472

Principles of method if other than guideline:

Gene mutation test was conducted on Salmonella typhimurium TA 100, TA 1535, TA 98, TA 15371 and Escherichia coli WP 2 uvr A 2 by using test chemical

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Divinylbenzene

EC Number:

215-325-5

EC Name:

Divinylbenzene

Cas Number:

1321-74-0

Molecular formula:

C10H10

IUPAC Name:

1,2-di(ethenyl)benzene

Details on test material:

- Name of test material: Divinylbenzene- IUPAC name: 1,2-di(ethenyl)benzene - Molecular formula: C10H10- Molecular weight: 131.1969 g/mol- Substance type: Organic- Physical state: No data - Purity: No data- Impurities (identity and concentrations): No data

Method

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Additional strain / cell type characteristics:

not specified

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Details on mammalian cell type (if applicable):

not specified

Additional strain / cell type characteristics:

not specified

Cytokinesis block (if used):

not specified

Metabolic activation:

with and without

Metabolic activation system:

Rat liver, induced with phenobarbital and 5,6-benzoflavone

Test concentrations with justification for top dose:

-S9 mix; 0, 0.781, 1.56, 3.13, 6.25, 12.5, 25.0, 50.0 μg/plate (TA1535(Test 1)); 0, 1.56 - 50.0 μg/plate(TA1535(Test 2)); 0, 3.13 - 100 μg/plate(TA100, TA98,TA1537); 0, 6.25 - 200 μg/plate(WP2 uvrA)+S9 mix; 0, 6.25 - 200 μg/plate(TA100, TA1 535, TA98, TA1537); 0, 15.6 - 500 μg/plate(WP2 uvrA)

Vehicle / solvent:

DMSO

Controls

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

9-aminoacridine

sodium azide

other: 2- (2-furyl) -3- (5-nitro-2-furyl) acrylamide and 2-aminoanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium; preincubationDURATION- Preincubation period: 20 minutes- Exposure duration: 48 hoursNUMBER OF REPLICATIONS: 2 METHODS OF SLIDE PREPARATION AND STAINING TECHNIQUE USED: 0.1 mL of the test substance preparation solution, 0.5 mL of the phosphate buffer solution (0.5 mL of the S 9 mix in the S9 mix addition test) and 0.1 mL of the test bacterium solution were mixed in a small test tube and preincubated at 37 ° C. for 20 minutes , Add 2 mL of top agar and mix and sonicate it on a synthetic medium flat plate.

Evaluation criteria:

Among the five test bacteria used, in the S9 mix-free test or S9 mix addition test of one or more test bacteria, the average value of the number of mutant colonies on the flat plate containing the test substance was 2 Fold or more, and when reproducibility and dose dependency were observed in the increase, the test substance was judged to be mutagenic (positive) in this test system.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium, other: TA100, TA1535, TA98, TA1537

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 WP2 uvr A

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

Remarks on result:

other: Non mutagenic

Applicant's summary and conclusion

Conclusions:

The test chemical did not induce gene mutation in Salmonella typhimurium TA100, TA1535, TA98, TA1537, Escherichia coli WP2 uvrA - in the presence and absence of rat liver 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 TA100, TA1535, TA98, TA1537, Escherichia coli WP2 uvrA with and without Rat liver, induced with phenobarbital and 5,6-benzoflavone activation system. 0.1 mL of the test substance preparation solution, 0.5 mL of the phosphate buffer solution (0.5 mL of the S 9 mix in the S9 mix addition test) and 0.1 mL of the test bacterium solution were mixed in a small test tube and preincubated at 37 ° C. for 20 minutes, Add 2 mL of top agar and mix and sonicate it on a synthetic medium flat plate. As a control group, a solvent to be used or several kinds of positive control substance solutions was used in place of the test substance preparation solution. The names and dosages of positive control substances used for each test bacterium are shown in each Table. Solvents and positive control groups were shared with other tests performed simultaneously. Cultivation was carried out at 37 ° C. for 48 hours. Toxicity was observed at 50.0 μg/plate (TA100, TA1535, TA1537) and 100 μg/plate(TA98, WP2 uvrA) without an S9 mix, and at 100 μg/plate(TA100, TA1535, TA98, TA1537) and 250 μg/plate(WP2 uvrA) with an S9 mix. The test chemical did not induce gene mutation in Salmonella typhimurium TA100, TA1535, TA98, TA1537, Escherichia coli WP2 uvrA - in the presence and absence of rat liver activation system and hence the chemical is not likely to classify as a gene mutant in vitro.