1-chloro-2,5-dimethoxy-4-nitrobenzene

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in mammalian cells

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1988-12-20 to 1989-01-26

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Well documented GLP and guideline study according to prior OECD TG.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1989

Report date:

1989

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)

Deviations:

no

GLP compliance:

yes

Remarks:

GLP compliance statement attached to full study report.

Type of assay:

mammalian cell gene mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

1-chloro-2,5-dimethoxy-4-nitrobenzene

EC Number:

230-086-7

EC Name:

1-chloro-2,5-dimethoxy-4-nitrobenzene

Cas Number:

6940-53-0

Molecular formula:

C8H8ClNO4

IUPAC Name:

1-chloro-2,5-dimethoxy-4-nitrobenzene

Method

Target gene:

Eucaryotic cells: Mutants at the HGPRT (hypoxanthine-guanine phosphoribosyl transferase) locus.

Species / strain

Species / strain / cell type:

Chinese hamster lung fibroblasts (V79)

Metabolic activation:

with and without

Metabolic activation system:

S9-Mix

Test concentrations with justification for top dose:

80, 300, 600, 800 µg/ml

Vehicle / solvent:

DMSO

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

ethylmethanesulphonate

Remarks:

without S9-mix

Migrated to IUCLID6: concentration: 1mg/ml

Controls 2

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9,10-dimethylbenzanthracene

Remarks:

with S9-Mix

Migrated to IUCLID6: 7.7 µg/ml dissolved in DMSO, final concentration: 0.5%

Details on test system and experimental conditions:

- Cell culture medium: MEM (Minimal essential medium) with Hanks salts and 25 mM Hepes-buffer
- Two days old exponentially growing cultures more than 50% confluent were trypsinated and a single cell suspension was prepared
- Subsequently, the cells were replated for mutagenicity testing and for determination of plating efficiency
- Treatment of the cells with the test substance in the presence or the absence of metabolic activation for 4 hrs

Evaluation criteria:

- The test substance is classified as mutagenic if it induces with one of the test substance concentrations reproducibly a mutation frequency that is three times higher than the spontaneous mutant frequency in this experiment.
- The test substance is classified as mutagenic if there is a reproducible concentration-related increase in the mutation frequency. Such evaluations may be considered independently from the enhancement factor for induced mutants.
However, in a case by case evaluation both decisions depend on the level of the corresponding negative control data.

Results and discussion

Test results

Species / strain:

Chinese hamster lung fibroblasts (V79)

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Remarks:

Tested up to the limit of solubility of 800 µg/ml.

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Additional information on results:

- Highest concentration w/o precipitation was found to be 800µg/mL
- The test item was proven not to be cytotoxic to the cells up to the limit of solubility
- 800 µg/mL was the highest dose tested in the experiments on mutagenicity

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

The results permit to draw the conclusion that the test compound is not mutagenic in the HGPRT-test with cells of the V79 Chinese hamster cell line.