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Toxicological information

Genetic toxicity: in vitro

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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

Referenceopen allclose all

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 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

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
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.