Hydrogen [2,4-dihydro-4-[(2-hydroxy-5-nitrophenyl)azo]-5-methyl-2-phenyl-3H-pyrazol-3-onato(2-)][2-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]benzoato(2-)]chromate(1-)

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:

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

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

Test material information

Constituent 1

Reference substance name:

4,4'-[(3,3'-dichloro[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[2,4-dihydro-5-methyl-2-phenyl-3H-pyrazol-3-one]

EC Number:

222-530-3

EC Name:

4,4'-[(3,3'-dichloro[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[2,4-dihydro-5-methyl-2-phenyl-3H-pyrazol-3-one]

Cas Number:

3520-72-7

Molecular formula:

C32H24Cl2N8O2

IUPAC Name:

4,4'-(1E,1'E)-(3,3'-dichlorobiphenyl-4,4'-diyl)bis(diazene-2,1-diyl)bis(3-methyl-1-phenyl-1H-pyrazol-5(4H)-one)

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/10^8viable 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.