Registration Dossier

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

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:
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:
Mutagenicity of 19 Major Graphic Arts And Printing Dyes
Author:
Paul Milvy, Kingsley Kay
Year:
1978
Bibliographic source:
Journal of Toxicology and Environmental Health, 4, 31, (1978)

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Principles of method if other than guideline:
Gene mutation study was conducted to evaluate the mutagenic nature of Diarylide orange
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material: Diarylide orange
- Other name: C.I. Pigment Orange 13
- Molecular formula: C32H24Cl2N8O2
- Molecular weight: 623.5016 g/mol
- Substance type: Organic
- Physical state: No data available
- Purity No data available
- Impurities (identity and concentrations): No data available
Specific details on test material used for the study:
- Name of test material: Diarylide orange
- Other name: C.I. Pigment Orange 13
- Molecular formula: C32H24Cl2N8O2
- Molecular weight: 623.5016 g/mol
- Substance type: Organic
- Physical state: No data available
- Purity No data available
- Impurities (identity and concentrations): No data available

Method

Target gene:
Histidine
Species / strain
Species / strain / cell type:
S. typhimurium, other: TA98, TA1538 and TA1535
Details on mammalian cell type (if applicable):
Not applicable
Additional strain / cell type characteristics:
other: Histidine auxotrophs
Cytokinesis block (if used):
No data
Metabolic activation:
with and without
Metabolic activation system:
Microsomal detoxifying enzymes using a 9000 Xg homogenate prepared from Aroclor 1254- induced male Swiss Webster mice
Test concentrations with justification for top dose:
0 or 10µg
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The test 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:
other: 2-aminofluorine, dimethylnitrosamine
Details on test system and experimental conditions:
METHOD OF APPLICATION: Preincubation

DURATION
- Preincubation period: 30 min
- Exposure duration: No data available
- Expression time (cells in growth medium): No data available
- Selection time (if incubation with a selection agent): No data available
- 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: No data available

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: No data available
Rationale for test conditions:
No data
Evaluation criteria:
An increase in the number of revertants was noted
Statistics:
No data

Results and discussion

Test results
Species / strain:
S. typhimurium, other: TA98, TA1538 and TA1535
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
Remarks on result:
other: No mutagenic potential

Any other information on results incl. tables

TABLE:Summary of Mutagenicity Test

Dye

Amount added

Condition

Revertants/108viable cells

TA1538

TA98

TA1535

Iron blue

0.01 mg dissolved in 0.01 ml DMSO

Complete Ames Assay

3.5 (2-8)b

-

0.8 (0-1.4)b

Iron blue

Same as above

No S9 homogenate

1.5±0.4

-

0.6 (0-1.6)b

Control

0.01 mL DMSO

Complete Ames Assay

3.9±2.2

1.1

0.4

Control

0.5 mg 2-aminofluorine

in 0.01 ml DMSO

Complete Ames Assay

1500 approx

-

-

Control

0.05 mg dimethylnitrosamine

In 0.01 ml DMSO

Complete Ames Assay

-

-

6000 approx

 

b: Mean (range)

Applicant's summary and conclusion

Conclusions:
Diarylide orange did not induce mutation in the Salmonella typhimurium TA98, TA1538 and TA1535 in the presence and absence of S9 metabolic activation system and hence is negative for gene mutation in vitro.
Executive summary:

Gene mutation study was conducted to evaluate the mutagenic nature of Diarylide orange. The study was performed using the preincubation protocol using Salmonella typhimurium TA98, TA1538 and TA1535 both in the presence and absence of S9 metabolic activation system.10 µg of the dye partially or completely dissolved in 0.01 ml of dimethyl sulfoxide (DMSO) was added to 0.9 ml of the reagents in the liquid phase and incubated 30 min at 37°C with shaking before plating 0.1 ml onto minimal plates. Diarylide orangedid not induce mutation in the Salmonella typhimurium TA98, TA1538 and TA1535 in the presence and absence of S9 metabolic activation system and hence is negative for gene mutation in vitro.