4-nitro-m-phenylenediamine

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:

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:

Phenazine derivatives as the mutagenic reaction product from o- or m-phenylenediamine derivatives with hydrogen peroxide

Author:

Tetsushi Watanabe, Teruhisa Hirayama and Shozo Fukui

Year:

1989

Bibliographic source:

Mutation Research, 227 (1989) 135-145

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Principles of method if other than guideline:

Gene mutation toxicity study was performed to determine the mutagenic nature of p-nitro-m-phenylenediamine (p-NO2-m-PD)

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

4-nitro-m-phenylenediamine

EC Number:

225-876-3

EC Name:

4-nitro-m-phenylenediamine

Cas Number:

5131-58-8

Molecular formula:

C6H7N3O2

IUPAC Name:

4-nitro-m-phenylenediamine

Test material form:

solid

Details on test material:

- Name of test material : 4-Nitro-m-phenylenediamine
- Molecular formula : C6H7N3O2
- Molecular weight : 153.14 g/mol
- Smiles notation : c1(c(cc(N)cc1)N)[N+](=O)[O-]
- InChl (if other than submission substance): 1S/C6H7N3O2/c7-4-1-2-6(9(10)11)5(8)3-4/h1-3H,7-8H2
- Substance type: Organic
- Physical state: Solid

Specific details on test material used for the study:

- Name of test material : 4-nitro-m-phenylenediamine
- IUPAC name: 4-nitrobenzene-1,3-diamine
- Molecular formula : C6H7N3O2
- Molecular weight : 153.14 g/mol
- Substance type: organic
- Physical state: No data
- Purity: No data available
- Impurities (identity and concentrations): No data available

Method

Target gene:

Histidine

Species / strain

Species / strain / cell type:

S. typhimurium TA 98

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

not specified

Cytokinesis block (if used):

No data

Metabolic activation:

with and without

Metabolic activation system:

The postmitochondrial fraction (S9) was prepared from the liver of male Sprague-Dawley rats induced with PCB.

Test concentrations with justification for top dose:

0, 10 or 30 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The chemical was soluble in 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:

2-acetylaminofluorene

4-nitroquinoline-N-oxide

Details on test system and experimental conditions:

METHOD OF APPLICATION: in suspension

DURATION
- Preincubation period: No data
- Exposure duration: No data
- Expression time (cells in growth medium): No data
- Selection time (if incubation with a selection agent): No data
- Fixation time (start of exposure up to fixation or harvest of cells): No data

SELECTION AGENT (mutation assays): No data
SPINDLE INHIBITOR (cytogenetic assays): No data
STAIN (for cytogenetic assays): No data

NUMBER OF REPLICATIONS: Triplicate

NUMBER OF CELLS EVALUATED: No data

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data

OTHER EXAMINATIONS:
- Determination of polyploidy: No data
- Determination of endoreplication: No data
- Other: No data

OTHER: No data

Rationale for test conditions:

No data

Evaluation criteria:

The plates were observed for a dose dependent increase in the number of revertants/plate

Statistics:

No data

Results and discussion

Test results

Species / strain:

S. typhimurium TA 98

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

Any other information on results incl. tables

Table: Mutagenicity for the test compoundp-nitro-m-phenylenediamine (p-NO2-m-PD)

Sample	Histidine revertants/dose
	-S9	+S9
p-NO2-m-PD	17/10	25/10
	28/30	32/30
DMSO	18	21
4NQO	411	-
AAF	-	470

Applicant's summary and conclusion

Conclusions:

p-nitro-m-phenylenediamine (p-NO2-m-PD) did not induce a dose dependent increase in the number of revertnats/plate as compared to controls using Salmonella typhimurium strain TA98 in the presence and absence of S9 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 p-nitro-m-phenylenediamine (p-NO2-m-PD). The study was performed by the Ames method with the suspension assay using Salmonella typhimurium strain TA98 with and without of S9 activation system. The test chemical dissolved in DMSO was used at dose levels of 0, 10 or 30 µg/plate in triplicate. Concurrent solvent and positive controls were included in the study.

 

p-nitro-m-phenylenediamine (p-NO2-m-PD) did not induce a dose dependent increase in the number of revertnats/plate as compared to controls using Salmonella typhimurium strain TA98 in the presence and absence of S9 activation system and hence the chemical is not likely to classify as a gene mutant in vitro.