1,4-bis[(4-butylphenyl)amino]-5,8-dihydroxyanthraquinone

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 publication

Data source

Reference

Reference Type:

publication

Title:

Mutagenicity Testing of Certified Food Colors and Related Azo, Xanthene And Triphenylmethane Dyes with the Salmonella/Microsome System

Author:

Joseph P. Brown, Gerald W. Roehm and Ronald J. Brown

Year:

1978

Bibliographic source:

Mutation Research, 56 (1978) 249-271

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 by the spot assay was performed for Sudan IV to evalute its mitagenic potential.

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

1-(2-methyl-4-(2-methylphenylazo)phenylazo)-2-naphthol

EC Number:

201-635-8

EC Name:

1-(2-methyl-4-(2-methylphenylazo)phenylazo)-2-naphthol

Cas Number:

85-83-6

Molecular formula:

C24H20N4O

IUPAC Name:

1-(2-methyl-4-(2-methylphenylazo)phenylazo)-2-naphthol / 85-83-6 / 201-635-8

Details on test material:

- Name of test material:C.I. Solvent Red 24
- Molecular formula :C24H20N40
- Molecular weight :380.48 g/mole
- Smiles notation: c12c(\N=N\c3c(cc(\N=N\c4c(cccc4)C)cc3)C)c(ccc1cccc2)O
- InChl: 1S/C24H20N4O/c1-16-7-3-6-10-21(16)26-25-19-12-13-22(17(2)15-19)27-28-24-20-9-5-4-8-18(20)11-14-23(24)29/h3-15,29H,1-2H3/b26-25+,28-27+
- Substance type:Organic

Specific details on test material used for the study:

- Name of test material: Sudan IV
- Molecular formula: C24H20N4O
- Molecular weight: 380.449 g/mol
- Substance type: Organic
- Physical state: No data
- Purity: No data
- Impurities (identity and concentrations): No data
- Smiles: c12c(\N=N\c3c(cc(\N=N\c4c(cccc4)C)cc3)C)c(ccc1cccc2)O

Method

Target gene:

Histidine

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA100, TA98, TA1535, TA1537 and TA1538

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:

S9 metabolic activation system isolated female Sprague Dawley rats given ip injection of Aroclor 1254 dissolved in corn oil

Test concentrations with justification for top dose:

200 µg/Plate

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:

not specified

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

9-aminoacridine

ethylmethanesulphonate

methylmethanesulfonate

other: Anthragallol, 2-anthramine

Details on test system and experimental conditions:

METHOD OF APPLICATION: Spot test

DURATION
- Preincubation period: No data
- Exposure duration: 72 hrs
- Expression time (cells in growth medium): 72 hrs
- 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: No data

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 criteria adopted for scoring a mutagenic response in routine plate tests was that the observed number of revertants should exceed twice the background value for that given assay and exceed the 99.9% confidence limit based on our historical controls.

Statistics:

No data

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA100, TA98, TA1535, TA1537 and TA1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Remarks on result:

other: No mutagenic effect were observed.

Any other information on results incl. tables

Table: Screening of Sudan IV

Test concentration	Chemical reduction (dithionite)	Microsome activation	Number of His^+revertants/plate
			TA1535	TA100	TA1537	TA1538	TA98
200µg/plate	-	-	-	-	-	-	-
		+	-	-	-	-	-

Applicant's summary and conclusion

Conclusions:

Sudan IV failed to induce mutation in Salmonella typhimurium strain TA100, TA98, TA1535, TA1537 and TA1538 with and without S9 metabolic activation system in the spot test performed and hence is not likely to classify for gene mutation in vitro.

Executive summary:

Gene mutation by the spot assay was performed for Sudan IV. Sudan IV was used at dose level of 200 µg/plate both with and without S9 metabolic activation system. The plates were incubated for 3 days at 37°C for the growth of colonies to occur. The plates were observed for an increase in the number of His+ revertants. The criteria adopted for scoring a mutagenic response in routine plate tests was that the observed number of revertants should exceed twice the background value for that given assay and exceed the 99.9% confidence limit based on our historical controls. Concurrent positive control was also included in the study.Sudan IV failed to induce mutation in Salmonellatyphimurium strain TA100, TA98, TA1535, TA1537 and TA1538 with and without S9 metabolic activation system in the spot test performed and hence is not likely to classify for gene mutation in vitro.