Disodium 3-hydroxy-4-[(4-methyl-2-sulphonatophenyl)azo]-2-naphthoate

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Gene mutation toxicity study was performed by F. RAFII et al. (Food and Chemical Toxicology, 1997) to determine the mutagenic nature of D&C Red 6 by bacterial reverse mutation assay. D&C Red No. 6 (5858-81-1) was assessed for its possible mutagenic potential. For this purpose AMES assay was performed on Salmonella typhimurium TA98 and TA100. The test material was exposed at the concentration of50-200 µg/plate in the presence and absence of S9. No mutagenic effects were observed. Therefore D&C Red No. 6 was considered to be non mutagenic in the Salmonella typhimurium TA98 and TA100 by AMES test. Hence the substance cannot be classified as genetox in vitro.

Link to relevant study records

Reference

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

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.

Qualifier:

according to guideline

Guideline:

other: As mention below

Principles of method if other than guideline:

To evaluate the mutagenic potential of D&C Red No. 6 in Salmonella typhimurium TA98 and TA100 by AMES test.

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Specific details on test material used for the study:

Name of the test chemical: disodium 4-[(E)-2-(4-methyl-2-sulfophenyl)diazen-1-yl]-3-oxidonaphthalene-2-carboxylateCommon Name used in study : D&C Red 6Molecular Formula: C18H12N2Na2O6SMolecular Weight: 430.3468 g/molInChI: 1S/C18H14N2O6S.2Na/c1-10-6-7-14(15(8-10)27(24,25)26)19-20-16-12-5-3-2-4-11(12)9-13(17(16)21)18(22)23;;/h2-9,21H,1H3,(H,22,23)(H,24,25,26);;/q;2*+1/p-2/b20-19+;;Substance Type: OrganicPhysical State: Solid

Target gene:

Histidine

Species / strain / cell type:

S. typhimurium, other: TA98 and TA100

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

not specified

Cytokinesis block (if used):

not specified

Metabolic activation:

with and without

Metabolic activation system:

The S-9 fraction (Organon Teknika, Durham, NC, USA) was from Sprague-Dawley rats induced with Aroclor-1254.

Test concentrations with justification for top dose:

0,50-200 µg/plate

Vehicle / solvent:

Vehicle- Vehicle(s)/solvent(s) used: Water

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

Water

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: DMSO, 1,6-dinitropyrene and benzo[a]pyrene

Details on test system and experimental conditions:

Details on test system and conditionsMETHOD OF APPLICATION: Plate incorporation method

Rationale for test conditions:

Not specified.

Evaluation criteria:

A positive result in the Ames test, which is defined as a reproducible, dose-related, at least twofold increase in the number of revertants over background was not observed with any of the azo dyes or their metabolites either before or after incubation with S-9.

Statistics:

Standard deviation was observed.

Key result

Species / strain:

S. typhimurium, other: TA98 and TA100.

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

Remarks on result:

other: No mutagenic effect were observed.

Table 1. Number of revertants of Salmonella typhimurium TAI00 with various azo dyes in the presence and absence of S-9 fraction

	Dye without S-9		Dye with S-9
	50 µg	200 µg	50 µg	200 µg
D&C Red No. 6	185 ± 15	111 ± 14	156 ± 8	127 ± 5

^a300 lag of dye was used.^bNot applicable.^C5 lag/plate of 1,6-dinitropyrene or benzo[a]pyrene was used.

Table 2. Number of revertants of Salmonella typhimurium TA98 with various azo dyes in the presence and absence of S-9 fraction

	Dye without S-9		Dye with S-9
	50 µg	200 µg	50 µg	200 µg
D&CRedNo. 6	33±4	34±8	27±11	17±4

^aNot applicable.^b5 lag/plate of 1,6-dinitropyrene or benzo[a]pyrene was used.

Conclusions:

D&C Red No. 6 (5858-81-1) was evaluated for its mutagenic potential in Salmonella typhimurium TA98 and TA100 by AMES test. The test result was considered to be negative in the presence and absence of metabolic activation.

Executive summary:

D&C Red No. 6 (5858-81-1) was assessed for its possible mutagenic potential. For this purpose AMES assay was performed on Salmonella typhimurium TA98 and TA100. The test material was exposed at the concentration of50-200 µg/plate in the presence and absence of S9. No mutagenic effects were observed. Therefore D&C Red No. 6 was considered to be non mutagenic in the Salmonella typhimurium TA98 and TA100 by AMES test. Hence the substance cannot be classified as genetox in vitro.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Genotoxicity In-vitro

Various publications were reviewed to determine the mutagenic nature of disodium 4-[(E)-2-(4-methyl-2-sulfophenyl)diazen-1-yl]-3-oxidonaphthalene-2-carboxylate /(5858-81-1) Other name: D&C Red 6 and Lithol rubine B(R201). The studies are as mentioned below:

Gene mutation toxicity study was performed by F. RAFII et al. (Food and Chemical Toxicology, 1997) to determine the mutagenic nature of D&C Red 6 by bacterial reverse mutation assay. D&C Red No. 6 (5858-81-1) was assessed for its possible mutagenic potential. For this purpose AMES assay was performed on Salmonella typhimurium TA98 and TA100. The test material was exposed at the concentration of50-200 µg/plate in the presence and absence of S9. No mutagenic effects were observed. Therefore D&C Red No. 6 was considered to be non mutagenic in the Salmonella typhimurium TA98 and TA100 by AMES test. Hence the substance cannot be classified as genetox in vitro.

Supported by a experimental study which is performed by J.P. BROWN et al. (Mutation Research, 1979) to determine the mutagenic nature of D&C Red 6. D and C Red No. 6 was assessed for its possible mutagenic potential. For this purpose bacterial reverse mutation assay was performed on Salmonella typhimurium strains TA1535, TA100, TA1537, TA1538 and TA98. The test material was exposed at the concentration of 0, 50,100and 500µg/plate in the presence and absence of S9 mix. Number of HIS + Revertants/plate was observed for test substance and compared with positive and negative control. No mutagenic effect were observed in any of the strain ,both in the presence and absence of S9.Therefore D and C Red No. 6 was considered to be non mutagenic in Salmonella typhimurium strains TA1535, TA100, TA1537, TA1538 and TA98 by bacterial reverse mutation assay. Hence the substance cannot be classified as gene mutant in vitro.

Another supporting study conducted by JEANNE M. MUZZALLet al. (Mutation Research, 1979) to determine the mutagenic nature of D&C Red 6(5858-81-1).D and C Red No. 6 was assessed for its possible mutagenic potential. For this purpose bacterial reverse mutation assay was performed on Salmonella typhimurium strains TA1535, TA100, TA1537and TA98. The test material was exposed at the concentration of 0, 10-250 mg in the presence and absence of S9 mix. Number of HIS + Revertants/plate was observed for test substance and compared with negative control. No mutagenic effect were observed in any of the strain ,both in the presence and absence of S9.Therefore D and C Red No. 6 was considered to be non mutagenic in Salmonella typhimurium strains TA1535, TA100, TA1537 and TA98 by bacterial reverse mutation assay. Hence the substance cannot be classified as gene mutant in vitro.

 

It is further supported by experimental study conducted by MIYAGOSHI M et al. (EISEI KAGAKU (J HYG CHEM), 1983) to determine the mutagenic nature of Lithol rubine B(R201) CAS NO; 5858-81-1. Lithol rubine B(R201)f was assessed for its possible mutagenic potential. For this purpose bacterial reverse mutation assay was performed on Salmonella typhimurium strains TA 100 and TA98. The test material was exposed at the concentration of 0, 50,100, 500 and 1000µg/plate in the presence and absence of S9 mix. Number of HIS + Revertants/plate was observed for test substance and compared with positive and negative control. No mutagenic effect were observed in any of the strain ,both in the presence and absence of S9.Therefore Lithol rubine B(R201)was considered to be non mutagenic in Salmonella typhimurium strains TA 100and TA98 by bacterial reverse mutation assay. Hence the substance cannot be classified as gene mutant in vitro.

 

Based on the data available for the target chemical disodium 4-[(E)-2-(4-methyl-2-sulfophenyl)diazen-1-yl]-3-oxidonaphthalene-2-carboxylate /(5858-81-1) Other name: D&C Red 6 and Lithol rubine B(R201) does not induce gene mutation in vitro. Hence the test chemical is not likely to classify as a gene mutant in vitro.

Justification for classification or non-classification

Thus based on the above annotation and CLP criteria for the target chemical .Disodium 4-[(E)-2-(4-methyl-2-sulfophenyl)diazen-1-yl]-3-oxidonaphthalene-2-carboxylate /(5858-81-1) Other name: D&C Red 6 and Lithol rubine B(R201) does not induce gene mutation in vitro. Hence the test chemical is not likely to classify as a gene mutant in vitro.