2-chloro-p-phenylenediamine

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:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Justification for type of information:

Data is from peer-reviewed journal.

Data source

Reference

Reference 1

Reference Type:

publication

Title:

Development of mutagenicity tests using Escherichia coli K-12 as indicator organism

Author:

Georges Mohn, Jürgen Ellenberger, Douglas McGregor

Year:

1974

Bibliographic source:

Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis, Volume 25, Issue 2, November 1974, Pages 187–196

Reference 2

Reference Type:

secondary source

Title:

OPINION ON 2-Chloro-p-phenylenediamine COLIPA n° A8

Author:

European Cosmetics Toiletry and Perfumery Association (COLIPA)

Year:

2013

Bibliographic source:

Scientific Committee on Consumer Safety (SCCS), SCCS/1510/13, 19 September 2013

Materials and methods

Test guideline

Qualifier:

no guideline available

Principles of method if other than guideline:

In –vitro forward and reverse bacterial gene mutation assay for 2-chloro-p-phenylenediamine was studied in E. coli.

GLP compliance:

not specified

Type of assay:

bacterial gene mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2-chloro-p-phenylenediamine

EC Number:

210-441-2

EC Name:

2-chloro-p-phenylenediamine

Cas Number:

615-66-7

Molecular formula:

C6H7ClN2

IUPAC Name:

2-chlorobenzene-1,4-diamine

Constituent 2

Reference substance name:

1,4 –diamino-2-chlorbenzol

IUPAC Name:

1,4 –diamino-2-chlorbenzol

Details on test material:

- Name of test material (as cited in study report): 1,4 –diamino-2-chlorbenzol
- Molecular formula (if other than submission substance): C6-H7-Cl-N2
- Molecular weight (if other than submission substance): 142.588 g/mol
- Substance type: Organic
- Physical state: Solid
- Purity: 99%
- Impurities (identity and concentrations): 1 %

Specific details on test material used for the study:

- Name of test material (as cited in study report): 1,4 –diamino-2-chlorbenzol
- Molecular formula: C6-H7-Cl-N2
- Molecular weight: 142.588 g/mol
- Substance type: Organic
- Physical state: Solid
- Purity: 99%
- Impurities (identity and concentrations): 1 %

Method

Target gene:

343/113

Species / strain

Species / strain / cell type:

E. coli, other: E. coli 343/113

Details on mammalian cell type (if applicable):

No data available

Additional strain / cell type characteristics:

not specified

Metabolic activation:

not specified

Test concentrations with justification for top dose:

1, 10 and 100 μg/ml

Vehicle / solvent:

Vehicle
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: No data available

Controls

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

not specified

True negative controls:

not specified

Positive controls:

not specified

Positive control substance:

not specified

Details on test system and experimental conditions:

Details on test system and conditions
METHOD OF APPLICATION: in agar (plate incorporation);

DURATION
- Preincubation period: No data available
- Exposure duration: 2 hours
- Expression time (cells in growth medium): 20 hours for standard medium, of 40 hours for gal+ mutants, and of 72 hours arg+ , nad+ and MTR mutants.
- Selection time (if incubation with a selection agent): 3 hours
- Fixation time (start of exposure up to fixation or harvest of cells): No data available

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

NUMBER OF REPLICATIONS: Duplicate cultures per “mutant” type in one experiment

NUMBER OF CELLS EVALUATED: No data available

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

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

OTHER: Both forward and reverse mutations in different genes: reverse mutations of arg- and nad- to prototrophy, forward mutations of 5-methyl-DL-tryptohan sensitivity (MTS) to MT resistance and forward (and reverse) mutations from Rs 18 to gal+.

Evaluation criteria:

A biologically relevant increase in the number of revertants

Statistics:

No data available

Results and discussion

Test results

Species / strain:

E. coli, other: E. coli 343/113

Metabolic activation:

not specified

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: No data available
- Effects of osmolality: No data available
- Evaporation from medium: No data available
- Water solubility: No data available
- Precipitation: No data available
- Other confounding effects: No data available

RANGE-FINDING/SCREENING STUDIES: Test concentrations were based on the basis of survival in a toxicity test with 3 h incubation with 1, 4–diamino-2-chlorbenzol in the dark.

COMPARISON WITH HISTORICAL CONTROL DATA: No data available

ADDITIONAL INFORMATION ON CYTOTOXICITY: No data available

Remarks on result:

other: no mutagenic potential

Applicant's summary and conclusion

Conclusions:

The substance 2-chloro-p-phenylenediamine (1,4–diamino-2-chlorbenzol) is non mutagenic to the bacterial tester strain E. coli 343/113.

Executive summary:

In a in –vitro forward and reverse bacterial gene mutation assay, E. coli343/113 was treated with 2-chloro-p-phenylenediamine (1,4–diamino-2-chlorbenzol) in concentration of 1, 10 and 100 μg/ml. No effect were observed on survival of E. coli343/113 during 2 hours incubation and an expression period of 20 hours for standard medium, of 40 hours for gal⁺mutants, and of 72 hours arg⁺, nad⁺and MTR mutants for forward mutations of 5-methyl-DL-tryptohan sensitivity (MTS) to MT resistance and forward (and reverse) mutations from Rs₁₈to gal⁺.

 

The substance 2-chloro-p-phenylenediamine (1,4–diamino-2-chlorbenzol) is non mutagenic to the bacterial tester strain E. coli 343/113.

 

According to the CLP classification, the test material does not classify as a gene mutant in vitro.