Dinitrotoluene

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Reliability as cited in OECD SIDS Dinitrotoluene (isomers mixture) CAS No.: 25321-14-6, 2004. Comparable to guideline study with acceptable restrictions; only one test concentration shown, no E.coli or TA102 tested

Data source

Reference

Reference 1

Reference Type:

publication

Title:

The mutagenicity of dinitrotoluenes in Salmonella typhimurium.

Author:

Couch B.D. et al.

Year:

1981

Bibliographic source:

Mutat. Res. 90, 373-383

Reference 2

Reference Type:

publication

Title:

The mutagenicity of dinitrotoluenes in Salmonella Typhimurium.

Author:

Couch D.B. et al.

Year:

1978

Bibliographic source:

Environ. Mutagen. 1, 168

Reference 3

Reference Type:

secondary source

Title:

Dinitrotoluene (isomers mixture), CAS No.: 25321-14-6

Author:

OECD SIDS

Year:

2004

Bibliographic source:

OECD SIDS

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

1000 µg(plate was the maximum concentration; only three concentrations tested; no positive controls used

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Dinitrotoluene

EC Number:

246-836-1

EC Name:

Dinitrotoluene

Cas Number:

25321-14-6

Molecular formula:

C7H6N2O4

IUPAC Name:

1-methyl-2,4-dinitrobenzene; 2-methyl-1,3-dinitrobenzene

Details on test material:

- Name of test material (as cited in study report): technical grade 2,4-dinitrotoluene (2,4-DNT)
- Isomers composition: 76.5 % 2,4-DNT (purity > 99.89%), 18.8 % 2.6-DNT (purity > 99.89%), 2.4 % 3.4-DNT (purity > 99.89%), 1.5 % 2.3-DNT (purity not confirmed), 0.7 % 2,5-DNT (purity not confirmed), 0.04 % 3,5-DNT (purity > 99.89%)

Method

Target gene:

his-

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA 98, TA 100, TA 1535, TA 1537, TA 1538

Metabolic activation:

with and without

Metabolic activation system:

S9-mix from the livers of Aroclor-treated (500 mg/kg, i.p., 5 days prior to sacrifice) male F344 rats

Test concentrations with justification for top dose:

3 concentrations up to 1000 µg/mL

Vehicle / solvent:

DMSO was used as solvent.

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

1% DMSO

True negative controls:

no

Positive controls:

not specified

Details on test system and experimental conditions:

METHOD OF APPLICATION: preincubation


DURATION
- Preincubation period: 3 h with and without S9
- Expression time: 40 h incubation at 37 °C


NUMBER OF REPLICATIONS: no data for preincubation test; n=3 for suspension assay


DETERMINATION OF CYTOTOXICITY
- Method: colony forming

Evaluation criteria:

two-fold increase of number of mutants/plate over control

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium, other: TA 1535, TA 1537, TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

other: Treatments which caused greater than 90% cytotoxicity relative to solvent controls were not employed.

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

not examined

Test results 2

Species / strain:

S. typhimurium, other: TA 1538, TA 98

Metabolic activation:

with and without

Genotoxicity:

positive

Cytotoxicity / choice of top concentrations:

other: Treatments which caused greater than 90% cytotoxicity relative to solvent controls were not employed.

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

not examined

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Results were only shown for the highest concentration tested.

		Revertants/plate
		TA1535		TA1537		TA1538		TA98		TA100
		-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9
2,4-DNT technical	1000 µg/plate	8	13	7	8	98	43	34	56	2	11
DMSO	1%	13.3 ± 7.9	8.1 ± 3.9	4.9 ± 3.0	5.1 ± 3.3	9.0 ± 6.5	15.3 ± 8.8	15.5 ± 4.5	24.8 ± 8.8	74.2 ± 33.5	63.1 ± 17.9

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
positive