4-aminoazobenzene-3,4'-disulphonic acid

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Available data suggests that the chemical 4-Aminoazobenzene-3,4'-disulphonic acid is negative for gene mutation in vitro and in vivo.

Link to relevant study records

Reference

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

(Q)SAR

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Justification for type of information:

QSAR prediction: migrated from IUCLID 5.6

Principles of method if other than guideline:

Prediction is done using QSAR Toolbox version 3.3

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Target gene:

No data

Species / strain / cell type:

S. typhimurium TA 100

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with

Metabolic activation system:

S9 activation system

Test concentrations with justification for top dose:

No data

Vehicle / solvent:

No data

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

Remarks:

No data

Details on test system and experimental conditions:

No data

Evaluation criteria:

No data

Statistics:

No data

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with

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:

No data

Remarks on result:

other: strain/cell type:

Remarks:

Migrated from field 'Test system'.

The prediction was based on dataset comprised from the following descriptors: "Gene mutation"
Estimation method: Takes highest mode value from the 5 nearest neighbours
Domain  logical expression:Result: In Domain

((((((((("a" or "b" or "c" or "d" or "e" )  and ("f" and ( not "g") )  )  and ("h" and ( not "i") )  )  and ("j" and ( not "k") )  )  and ("l" and ( not "m") )  )  and ("n" and ( not "o") )  )  and "p" )  and "q" )  and ("r" and "s" )  )

Domain logical expression index: "a"

Referential boundary: The target chemical should be classified as Anilines (Acute toxicity) by US-EPA New Chemical Categories

Domain logical expression index: "b"

Referential boundary: The target chemical should be classified as Aniline AND Aryl AND Azo AND Sulfonic acid by Organic Functional groups

Domain logical expression index: "c"

Referential boundary: The target chemical should be classified as Aniline AND Aryl AND Azo AND Overlapping groups AND Sulfonic acid by Organic Functional groups (nested)

Domain logical expression index: "d"

Referential boundary: The target chemical should be classified as Aliphatic Nitrogen, one aromatic attach [-N] AND Aromatic Carbon [C] AND Azo [-N=N-] AND Hydroxy, sulfur attach [-OH] AND Miscellaneous sulfide (=S) or oxide (=O) AND Olefinic carbon [=CH- or =C<] AND Suflur {v+4} or {v+6} AND Sulfinic acid [-S(=O)OH] AND Sulfonate, aromatic attach [-SO2-O] by Organic functional groups (US EPA)

Domain logical expression index: "e"

Referential boundary: The target chemical should be classified as Amine AND Aromatic compound AND Azo compound AND Primary amine AND Primary aromatic amine AND Sulfonic acid AND Sulfonic acid derivative by Organic functional groups, Norbert Haider (checkmol)

Domain logical expression index: "f"

Referential boundary: The target chemical should be classified as No alert found by DNA binding by OASIS v.1.3

Domain logical expression index: "g"

Referential boundary: The target chemical should be classified as AN2 OR AN2 >>  Michael-type addition, quinoid structures OR AN2 >>  Michael-type addition, quinoid structures >> Flavonoids OR AN2 >>  Michael-type addition, quinoid structures >> Quinoneimines OR AN2 >>  Michael-type addition, quinoid structures >> Quinones OR Non-covalent interaction OR Non-covalent interaction >> DNA intercalation OR Non-covalent interaction >> DNA intercalation >> Acridone, Thioxanthone, Xanthone and Phenazine Derivatives OR Non-covalent interaction >> DNA intercalation >> Amino Anthraquinones OR Non-covalent interaction >> DNA intercalation >> Aminoacridine DNA Intercalators OR Non-covalent interaction >> DNA intercalation >> DNA Intercalators with Carboxamide Side Chain OR Non-covalent interaction >> DNA intercalation >> Fused-Ring Nitroaromatics OR Non-covalent interaction >> DNA intercalation >> Fused-Ring Primary Aromatic Amines OR Non-covalent interaction >> DNA intercalation >> Quinones OR Non-specific OR Non-specific >> Incorporation into DNA/RNA, due to structural analogy with  nucleoside bases    OR Non-specific >> Incorporation into DNA/RNA, due to structural analogy with  nucleoside bases    >> Specific Imine and Thione Derivatives OR Radical OR Radical >> Generation of reactive oxygen species OR Radical >> Generation of reactive oxygen species >> Thiols OR Radical >> Radical mechanism by ROS formation OR Radical >> Radical mechanism by ROS formation >> Acridone, Thioxanthone, Xanthone and Phenazine Derivatives OR Radical >> Radical mechanism by ROS formation >> Polynitroarenes OR Radical >> Radical mechanism via ROS formation (indirect) OR Radical >> Radical mechanism via ROS formation (indirect) >> Amino Anthraquinones OR Radical >> Radical mechanism via ROS formation (indirect) >> Flavonoids OR Radical >> Radical mechanism via ROS formation (indirect) >> Fused-Ring Nitroaromatics OR Radical >> Radical mechanism via ROS formation (indirect) >> Fused-Ring Primary Aromatic Amines OR Radical >> Radical mechanism via ROS formation (indirect) >> Hydrazine Derivatives OR Radical >> Radical mechanism via ROS formation (indirect) >> Nitro Azoarenes OR Radical >> Radical mechanism via ROS formation (indirect) >> Nitroaniline Derivatives OR Radical >> Radical mechanism via ROS formation (indirect) >> Nitroarenes with Other Active Groups OR Radical >> Radical mechanism via ROS formation (indirect) >> Nitrophenols, Nitrophenyl Ethers and Nitrobenzoic Acids OR Radical >> Radical mechanism via ROS formation (indirect) >> p-Aminobiphenyl Analogs OR Radical >> Radical mechanism via ROS formation (indirect) >> Quinones OR Radical >> Radical mechanism via ROS formation (indirect) >> Single-Ring Substituted Primary Aromatic Amines OR Radical >> Radical mechanism via ROS formation (indirect) >> Specific Imine and Thione Derivatives OR Radical >> ROS formation after GSH depletion (indirect) OR Radical >> ROS formation after GSH depletion (indirect) >> Quinoneimines OR SN1 OR SN1 >> Alkylation after metabolically formed carbenium ion species OR SN1 >> Alkylation after metabolically formed carbenium ion species >> Polycyclic Aromatic Hydrocarbon Derivatives OR SN1 >> Nucleophilic attack after carbenium ion formation OR SN1 >> Nucleophilic attack after carbenium ion formation >> Acyclic Triazenes OR SN1 >> Nucleophilic attack after diazonium or carbenium ion formation OR SN1 >> Nucleophilic attack after diazonium or carbenium ion formation >> Nitroarenes with Other Active Groups OR SN1 >> Nucleophilic attack after metabolic nitrenium ion formation OR SN1 >> Nucleophilic attack after metabolic nitrenium ion formation >> Amino Anthraquinones OR SN1 >> Nucleophilic attack after metabolic nitrenium ion formation >> Fused-Ring Primary Aromatic Amines OR SN1 >> Nucleophilic attack after metabolic nitrenium ion formation >> p-Aminobiphenyl Analogs OR SN1 >> Nucleophilic attack after metabolic nitrenium ion formation >> Single-Ring Substituted Primary Aromatic Amines OR SN1 >> Nucleophilic attack after reduction and nitrenium ion formation OR SN1 >> Nucleophilic attack after reduction and nitrenium ion formation >> Fused-Ring Nitroaromatics OR SN1 >> Nucleophilic attack after reduction and nitrenium ion formation >> Nitro Azoarenes OR SN1 >> Nucleophilic attack after reduction and nitrenium ion formation >> Nitroaniline Derivatives OR SN1 >> Nucleophilic attack after reduction and nitrenium ion formation >> Nitroarenes with Other Active Groups OR SN1 >> Nucleophilic attack after reduction and nitrenium ion formation >> Nitrobiphenyls and Bridged Nitrobiphenyls OR SN1 >> Nucleophilic attack after reduction and nitrenium ion formation >> Nitrophenols, Nitrophenyl Ethers and Nitrobenzoic Acids OR SN1 >> Nucleophilic attack after reduction and nitrenium ion formation >> Polynitroarenes OR SN1 >> Nucleophilic substitution on diazonium ions OR SN1 >> Nucleophilic substitution on diazonium ions >> Specific Imine and Thione Derivatives OR SN2 OR SN2 >> Alkylation, direct acting epoxides and related after P450-mediated metabolic activation OR SN2 >> Alkylation, direct acting epoxides and related after P450-mediated metabolic activation >> Polycyclic Aromatic Hydrocarbon Derivatives OR SN2 >> Direct acting epoxides formed after metabolic activation OR SN2 >> Direct acting epoxides formed after metabolic activation >> Quinoline Derivatives OR SN2 >> DNA alkylation OR SN2 >> DNA alkylation >> Alkylphosphates, Alkylthiophosphates and Alkylphosphonates OR SN2 >> SN2 at an activated carbon atom OR SN2 >> SN2 at an activated carbon atom >> Quinoline Derivatives OR SN2 >> SN2 attack on activated carbon Csp3 or Csp2 OR SN2 >> SN2 attack on activated carbon Csp3 or Csp2 >> Nitroarenes with Other Active Groups by DNA binding by OASIS v.1.3

Domain logical expression index: "h"

Referential boundary: The target chemical should be classified as Not possible to classify according to these rules by DPRA Cysteine peptide depletion

Domain logical expression index: "i"

Referential boundary: The target chemical should be classified as Low reactive OR Low reactive >> N-substituted aromatic amides OR Low reactive >> Sulfanilic acid derivatives by DPRA Cysteine peptide depletion

Domain logical expression index: "j"

Referential boundary: The target chemical should be classified as Strong binder, NH2 group by Estrogen Receptor Binding

Domain logical expression index: "k"

Referential boundary: The target chemical should be classified as Moderate binder, NH2 group OR Moderate binder, OH grooup OR Non binder, impaired OH or NH2 group OR Non binder, MW>500 OR Non binder, non cyclic structure OR Non binder, without OH or NH2 group OR Strong binder, OH group OR Weak binder, NH2 group OR Weak binder, OH group by Estrogen Receptor Binding

Domain logical expression index: "l"

Referential boundary: The target chemical should be classified as No alert found by Protein binding by OECD

Domain logical expression index: "m"

Referential boundary: The target chemical should be classified as Acylation OR Acylation >> Direct Acylation Involving a Leaving group OR Acylation >> Direct Acylation Involving a Leaving group >> Acetates OR SN2 OR SN2 >> SN2 reaction at sp3 carbon atom OR SN2 >> SN2 reaction at sp3 carbon atom >> Allyl acetates and related chemicals by Protein binding by OECD

Domain logical expression index: "n"

Referential boundary: The target chemical should be classified as Not possible to classify according to these rules (GSH) by Protein binding potency

Domain logical expression index: "o"

Referential boundary: The target chemical should be classified as Highly reactive (GSH) OR Highly reactive (GSH) >> Furamates (MA) by Protein binding potency

Domain logical expression index: "p"

Referential boundary: The target chemical should be classified as No superfragment by Superfragments ONLY

Domain logical expression index: "q"

Similarity boundary:Target: Nc1ccc(N=Nc2ccc(S(O)(=O)=O)cc2)cc1S(O)(=O)=O
Threshold=10%,
Dice(Atom centered fragments)
Atom type; Count H attached; Hybridization

Domain logical expression index: "r"

Parametric boundary:The target chemical should have a value of log Kow which is >= -2.18

Domain logical expression index: "s"

Parametric boundary:The target chemical should have a value of log Kow which is <= -0.598

Conclusions:

Interpretation of results (migrated information):negative with metabolic activationThe test material 4-Aminoazobenzene-3,4'-disulphonic acid is not mutagenic in vitro in Salmonella typhimurium strain TA100 with S9 metabolic activation system.

Executive summary:

Gene mutation was predicted using SSS QSAR prediction model, 2016. The study used Salmonella typhimurium TA100 strain with S9 activation system. The test material 4-Aminoazobenzene-3,4'-disulphonic acid is not mutagenic in vitro in Salmonella typhimurium strain TA100 with S9 metabolic activation system.

Hence the test material 4-Aminoazobenzene-3,4'-disulphonic acid is not mutagenic in vitro.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

Additional information from genetic toxicity in vitro:

Gene mutation in vitro:

Prediction model based estimation for the target chemical and data from read across have been summarized as below to determine the mutagenic nature of the test compound 4-Amino-3,4'-disulfoazobenzene.

Gene mutation was predicted using SSS QSAR prediction model, 2016. The study used Salmonella typhimurium TA100 strain with S9 activation system. The test material 4-Aminoazobenzene-3,4'-disulphonic acid is not mutagenic in vitro in Salmonella typhimurium strain TA100 with S9 metabolic activation system.

Gene mutation was predicted using SSS QSAR prediction model, 2016. The study used Salmonella typhimurium strain TA102 without S9 metabolic activation system. The test material 4-Aminoazobenzene-3,4'-disulphonic acid is not mutagenic in vitro in Salmonella typhimurium strain TA102 without S9 activation system.

Based on the QSAR prediction done using the Danish (Q)SAR Database (2016), the genetic toxicity was estimated to be negative during Ames test in S. typhimurium TA 1535, TA 1537, TA 98 and TA 100. Thus it can be concluded that the substance 4-aminoazobenzene-3, 4’-disulphonic acid is non mutagenic and also based on the CLP criteria for classification it can be not classified for genetic toxicity.

Based on the QSAR prediction done using the Danish (Q)SAR Database (2016), the genetic toxicity was estimated to be negative during Chromosome Aberrations in Chinese Hamster Ovary (CHO) Cells. Thus it can be concluded that the substance 4-aminoazobenzene-3, 4’-disulphonic acid is non mutagenic and also based on the CLP criteria for classification it can be not classified for genetic toxicity.

In a in vitro gene mutation test (Mansour, 2009), Salmonella typhimurium TA102 and TA104 was tested for Revertant bacterial colonies by using Acid orange 52 (AO52) with and without S9 activation in the concentration of 250, 50 and 10 g/plate. Revertant bacterial colonies were not observed in without S9 activation and with S9 metabolic activation revertant bacterial colonies were observed. Therefore, Acid orange 52 (AO52) was considered to be negative without S9 and positive gene toxic with S9 metabolic activation when Salmonella typhimurium TA102 and TA104 tested.

Ames mutagenicity assay was performed by Das et al (2004) to evaluate the mutagenic nature of the test compound tartrazine (RA CAS no 1934 -21 -0) in plate incorporation assay. The test material was tested at a concentration of 10, 100, 250, 500 and 1000 μg /plate. The plates were inverted within an hour and placed in a dark vented incubator at 37⁰C for 48 hours. Positive controls (for TA97a and TA98, 20 μg/plate nitro phenylene diamine and for TA100, 1.5 μg/plate sodium azide) and negative controls were maintained concurrently for all the experiments. Three plates were used for each set. After 48 hours of incubation, the revertant colonies were counted. The test compound tartrazine is not mutagenic in the study conducted using Salmonella typhimurium TA97a, TA98 and TA100 without metabolic activation system.  

Chromosomal aberration test was performed by Ishidate et al (1984) for the test chemical tartrazine (RA CAS no 1934 -21 -0) using Chinese hamster fibroblast cell line CHL. The cells were exposed to the test material at three different doses for 24 and 48 hr. In the present studies, no metabolic activation systems were applied. The incidence of polyploid cells as well as of cells with structural chromosomal aberrations such as chromatid or chromosome gaps, breaks, exchanges, ring formations, fragmentations and others, was recorded on each culture plate. Untreated cells and solvent-treated cells served as negative controls, in which the incidence of aberrations was usually less than 3.0%. The results were considered to be negative if the incidence was less than 4.9%, equivocal if it was between 5.0 and 9.9%, and positive if it was more than 10.0%. The incidence of chromosome aberration in Chinese hamster fibroblast cell line for the test chemical tartrazine is more than 10%. Hence tartrazine is mutagenic in vitro.

Salmonella typhimurium strains TA92, TA1535, TA100, TA1537, TA94 and TA98 were subjected to six concentrations of tartrazine (RA CAS no 1934 -21 -0) with the maximum concentration being 5000 µg/plate in the presence of S9 metabolic activation system (Ishidate et al, 1984). Cells cultured overnight were pre-incubated with both the test sample and the S-9 mix for 20 min at 37°C before plating. Duplicate plates were used for each of six different concentrations of the sample. The number of revertant (his +) colonies was scored after incubation at 37°C for 2 days. Tartrazine did not induce reversion of mutation when applied to Salmonella typhimurium strains in plate incorporation method. Gene toxicity in vitro is negative for tartrazine.

Gene mutation in vivo:

Gene mutation in vivo was predicted using SSS QSAR prediction model, 2016. Chromosome abberration study was performed for the test chemical 4-Aminoazobenzene-3,4'-disulphonic acid (CAS no 101-50-8) in the in vivo micronucleus assay using mice. The test chemical failed to induce mutation and hence is likely to be not classified for gene mutation in vivo.

 

Chromosome aberration assay was performed by Das et al (2004) to evaluate the mutagenic nature of the test compound tartrazine (RA CAS no 1934-21-0). The study was conducted on male Swiss albino mice, 8-10 weeks old and weighing 20-25 g. Four animals per dose were administered intraperitoneally with the different doses of the tartrazine (50,100 and 200 mg/kg body weight). The animals were killed after 18 hr. For bone marrow chromosome analysis, animals were injected with 0.1 ml colchicine solution (4mg/10ml distilled water /10g body weight, 90 minutes before they were killed. Bone marrow cells were routinely processed by the standard procedure and slides were coded and stained in diluted Giemsa. For chromosomal aberration analysis, four animals were used per point. Hundred well spread metaphase plates were scored per animal (400 metaphase plates per treatment set) at random. The types of aberrations were scored. The percentages of damaged cells (% DC) and chromosomal aberrations per cell (CA/cell) values were calculated excluding gaps. The test compound tartrazine is not mutagenic in the chromosomal aberration study conducted using Swiss albino mouse isolated bone marrow cells.

The food dye tartrazine (RA CAS 1934-21-0 ) was administered twice, at 24 h intervals, by oral gavage to mice and assessed in the in vivo gut micronucleus test for genotoxic effects (frequency of micronucleated cells) and toxicity (apoptotic and mitotic cells) (Poul et al, 2009). Acute oral exposure to food dye additives tartrazine at a concentration of 0, 20, 200 or 1000 mg/Kg bw did not induce genotoxic effect in the micronucleus gut assay in mice. Tartrazine however increased the mitotic cells at all dose levels when compared to controls. The test compound tartrazine is not genotoxic in vivo as per the results obtained from gut micronucleus assay using Swice albino mice.

 

Based on the predicted data for the target cas no and data from read across, the test chemical is likely to be not classified for gene mutation in vitro and in vivo.

Justification for selection of genetic toxicity endpoint
Gene mutation was predicted using SSS QSAR prediction model, 2016.

Justification for classification or non-classification

Weight of evidence data suggests that the chemical 4-Aminoazobenzene-3,4'-disulphonic acid is not likely to classify for gene mutation in vitro and in vivo.

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