4-(phenylazo)benzene-1,3-diamine acetate

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Gene mutation in vitro:

Ames test:

Based on the prediction done using the OECD QSAR toolbox version 3.3 with log kow as the primary descriptor and considering the five closest read across substances, gene mutation was predicted for 4-(phenylazo)benzene-1,3-diamine acetate. The study assumed the use of Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and TA 102 with S9 metabolic activation system. 4-(phenylazo)benzene-1,3-diamine acetate was predicted to induce gene mutation in Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and TA 102 in the presence of S9 metabolic activation system and hence, according to the prediction made, it is likely to classify as a gene mutant in vitro.

 

Based on the predicted result it can be concluded that the substance is considered to be mutagenic as per the criteria mentioned in CLP regulation.

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)

Rationale for reliability incl. deficiencies:

results derived from a valid (Q)SAR model and falling into its applicability domain, with limited documentation / justification

Justification for type of information:

Data is from OECD QSAR Toolbox version 3.3 and the supporting QMRF report has been attached

Qualifier:

according to guideline

Guideline:

other: Refer below principle

Principles of method if other than guideline:

Prediction is done using OECD QSAR Toolbox version 3.3, 2018

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Specific details on test material used for the study:

- Name of test material (IUPAC name): 4-(phenylazo)benzene-1,3-diamine acetate
- IUPAC name: 4-(phenylazo)benzene-1,3-diamine acetate
- Molecular formula: C12H12N4xC2H4O2
- Molecular weight: 272.306 g/mol
- Smiles notation: c1(ccc(c(N)c1)\N=N\c1ccccc1)N.C(C)(=O)O
- InChl: 1S/C12H12N4.C2H4O2/c13-9-6-7-12(11(14)8-9)16-15-10-4-2-1-3-5-10;1-2(3)4/h1-8H,13-14H2;1H3,(H,3,4)/b16-15+;
- Substance type: Organic

Target gene:

Histidine

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

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

Metabolic activation system:

S9 metabolic 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

Details on test system and experimental conditions:

No data

Rationale for test conditions:

No data

Evaluation criteria:

Prediction is done considering a dose dependent increase in the number of revertants/plate

Statistics:

No data

Species / strain:

S. typhimurium, other: TA 1535, TA 1537, TA 98 and TA 100

Metabolic activation:

with

Genotoxicity:

positive

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:

mutagenic potential (based on QSAR/QSPR prediction)

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" )  and "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 ( not "q") )  )  and ("r" and ( not "s") )  )  and ("t" and ( not "u") )  )  and ("v" and ( not "w") )  )  and "x" )  and ("y" and ( not "z") )  )  and ("aa" and ( not "ab") )  )  and ("ac" and ( not "ad") )  )  and ("ae" and ( not "af") )  )  and ("ag" and ( not "ah") )  )  and ("ai" and ( not "aj") )  )  and ("ak" and ( not "al") )  )  and ("am" and ( not "an") )  )  and ("ao" and ( not "ap") )  )  and ("aq" and ( not "ar") )  )  and ("as" and ( not "at") )  )  and ("au" and ( not "av") )  )  and ("aw" and ( not "ax") )  )  and ("ay" and "az" )  )

Domain logical expression index: "a"

Referential boundary: The target chemical should be classified as Anilines (Acute toxicity) AND Not categorized by US-EPA New Chemical Categories

Domain logical expression index: "b"

Referential boundary: The target chemical should be classified as SN1 AND SN1 >> Nitrenium Ion formation AND SN1 >> Nitrenium Ion formation >> Aromatic azo AND SN1 >> Nitrenium Ion formation >> Primary aromatic amine by DNA binding by OECD

Domain logical expression index: "c"

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

Domain logical expression index: "d"

Referential boundary: The target chemical should be classified as Anilines (amino-meta) AND Anilines (Unhindered) by Aquatic toxicity classification by ECOSAR

Domain logical expression index: "e"

Referential boundary: The target chemical should be classified as Bioavailable by Lipinski Rule Oasis ONLY

Domain logical expression index: "f"

Referential boundary: The target chemical should be classified as Non-Metals by Groups of elements

Domain logical expression index: "g"

Referential boundary: The target chemical should be classified as Alkali Earth OR Halogens OR Metalloids OR Transition Metals OR Unknown chemical element by Groups of elements

Domain logical expression index: "h"

Referential boundary: The target chemical should be classified as Group 14 - Carbon C AND Group 15 - Nitrogen N AND Group 16 - Oxygen O by Chemical elements

Domain logical expression index: "i"

Referential boundary: The target chemical should be classified as Group 15 - Phosphorus P OR Group 16 - Sulfur S by Chemical elements

Domain logical expression index: "j"

Referential boundary: The target chemical should be classified as Aliphatic acids and (Met)acrylic acids AND Aromatic amines by Skin irritation/corrosion Inclusion rules by BfR

Domain logical expression index: "k"

Referential boundary: The target chemical should be classified as Phenols by Skin irritation/corrosion Inclusion rules by BfR

Domain logical expression index: "l"

Referential boundary: The target chemical should be classified as Aliphatic acids and (Met)acrylic acids AND Aromatic amines by Skin irritation/corrosion Inclusion rules by BfR

Domain logical expression index: "m"

Referential boundary: The target chemical should be classified as Ketones by Skin irritation/corrosion Inclusion rules by BfR

Domain logical expression index: "n"

Referential boundary: The target chemical should be classified as Aliphatic acids and (Met)acrylic acids AND Aromatic amines by Skin irritation/corrosion Inclusion rules by BfR

Domain logical expression index: "o"

Referential boundary: The target chemical should be classified as Ethylenglycolethers by Skin irritation/corrosion Inclusion rules by BfR

Domain logical expression index: "p"

Referential boundary: The target chemical should be classified as Aromatic diazo AND H-acceptor-path3-H-acceptor AND Primary aromatic amine, hydroxyl amine and its derived esters by in vivo mutagenicity (Micronucleus) alerts by ISS

Domain logical expression index: "q"

Referential boundary: The target chemical should be classified as 1,3-dialkoxy-benzene OR 1-phenoxy-benzene by in vivo mutagenicity (Micronucleus) alerts by ISS

Domain logical expression index: "r"

Referential boundary: The target chemical should be classified as Aromatic diazo AND H-acceptor-path3-H-acceptor AND Primary aromatic amine, hydroxyl amine and its derived esters by in vivo mutagenicity (Micronucleus) alerts by ISS

Domain logical expression index: "s"

Referential boundary: The target chemical should be classified as Aromatic N-acyl amine OR Azide and triazene groups by in vivo mutagenicity (Micronucleus) alerts by ISS

Domain logical expression index: "t"

Referential boundary: The target chemical should be classified as Aromatic diazo AND H-acceptor-path3-H-acceptor AND Primary aromatic amine, hydroxyl amine and its derived esters by in vivo mutagenicity (Micronucleus) alerts by ISS

Domain logical expression index: "u"

Referential boundary: The target chemical should be classified as Heterocyclic Polycyclic Aromatic Hydrocarbons by in vivo mutagenicity (Micronucleus) alerts by ISS

Domain logical expression index: "v"

Referential boundary: The target chemical should be classified as Aromatic diazo AND No alert found AND Primary aromatic amine,hydroxyl amine and its derived esters by in vitro mutagenicity (Ames test) alerts by ISS

Domain logical expression index: "w"

Referential boundary: The target chemical should be classified as Polycyclic Aromatic Hydrocarbons by in vitro mutagenicity (Ames test) alerts by ISS

Domain logical expression index: "x"

Similarity boundary:Target: CC(O)=O_Nc1ccc(N=Nc2ccccc2)c(N)c1
Threshold=100%,
Dice(Atom centered fragments)
Atom type; Count H attached; Hybridization

Domain logical expression index: "y"

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

Domain logical expression index: "z"

Referential boundary: The target chemical should be classified as Alkylarylether by Organic functional groups, Norbert Haider (checkmol)

Domain logical expression index: "aa"

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

Domain logical expression index: "ab"

Referential boundary: The target chemical should be classified as Carboxylic acid amide by Organic functional groups, Norbert Haider (checkmol)

Domain logical expression index: "ac"

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

Domain logical expression index: "ad"

Referential boundary: The target chemical should be classified as Carboxylic acid ester by Organic functional groups, Norbert Haider (checkmol)

Domain logical expression index: "ae"

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

Domain logical expression index: "af"

Referential boundary: The target chemical should be classified as Carboxylic acid sec. amide by Organic functional groups, Norbert Haider (checkmol)

Domain logical expression index: "ag"

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

Domain logical expression index: "ah"

Referential boundary: The target chemical should be classified as Primary aliphatic amine by Organic functional groups, Norbert Haider (checkmol)

Domain logical expression index: "ai"

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

Domain logical expression index: "aj"

Referential boundary: The target chemical should be classified as Primary alcohol by Organic functional groups, Norbert Haider (checkmol)

Domain logical expression index: "ak"

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

Domain logical expression index: "al"

Referential boundary: The target chemical should be classified as Hydroxy compound OR Nitrile by Organic functional groups, Norbert Haider (checkmol)

Domain logical expression index: "am"

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

Domain logical expression index: "an"

Referential boundary: The target chemical should be classified as Tertiary aliphatic amine by Organic functional groups, Norbert Haider (checkmol)

Domain logical expression index: "ao"

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

Domain logical expression index: "ap"

Referential boundary: The target chemical should be classified as Secondary aromatic amine by Organic functional groups, Norbert Haider (checkmol)

Domain logical expression index: "aq"

Referential boundary: The target chemical should be classified as Acetoxy AND Aminoaniline, meta AND Aniline AND Aryl AND Azo AND Carboxylic acid by Organic Functional groups

Domain logical expression index: "ar"

Referential boundary: The target chemical should be classified as Alkene by Organic Functional groups

Domain logical expression index: "as"

Referential boundary: The target chemical should be classified as Acetoxy AND Aminoaniline, meta AND Aniline AND Aryl AND Azo AND Carboxylic acid by Organic Functional groups

Domain logical expression index: "at"

Referential boundary: The target chemical should be classified as Fused heterocyclic aromatic by Organic Functional groups

Domain logical expression index: "au"

Referential boundary: The target chemical should be classified as Acetoxy AND Aminoaniline, meta AND Aniline AND Aryl AND Azo AND Carboxylic acid by Organic Functional groups

Domain logical expression index: "av"

Referential boundary: The target chemical should be classified as Triazine by Organic Functional groups

Domain logical expression index: "aw"

Referential boundary: The target chemical should be classified as Acetoxy AND Aminoaniline, meta AND Aniline AND Aryl AND Azo AND Carboxylic acid by Organic Functional groups

Domain logical expression index: "ax"

Referential boundary: The target chemical should be classified as Precursors quinoid compounds by Organic Functional groups

Domain logical expression index: "ay"

Parametric boundary:The target chemical should have a value of log Kow which is >= 0.248

Domain logical expression index: "az"

Parametric boundary:The target chemical should have a value of log Kow which is <= 1.77

Conclusions:

4-(phenylazo)benzene-1,3-diamine acetate was predicted to induce gene mutation in Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and TA 102 in the presence of S9 metabolic activation system and hence, according to the prediction made, it is likely to classify as a gene mutant in vitro.

Executive summary:

Based on the prediction done using the OECD QSAR toolbox version 3.3 with log kow as the primary descriptor and considering the five closest read across substances, gene mutation was predicted for 4-(phenylazo)benzene-1,3-diamine acetate. The study assumed the use of Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and TA 102 with S9 metabolic activation system. 4-(phenylazo)benzene-1,3-diamine acetate was predicted to induce gene mutation in Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and TA 102 in the presence of S9 metabolic activation system and hence, according to the prediction made, it is likely to classify as a gene mutant in vitro.

 

Based on the predicted result it can be concluded that the substance is considered to be mutagenic as per the criteria mentioned in CLP regulation.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (positive)

Genetic toxicity in vivo

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Gene mutation in vitro:

Prediction model based estimation for the target chemical and data from read across chemicals have been reviewed to determine the mutagenic nature of 4-(phenylazo)benzene-1,3-diamine acetate (CAS no 79234 -33 -6). The studies are as mentioined below:

Based on the prediction done using the OECD QSAR toolbox version 3.3 with log kow as the primary descriptor and considering the five closest read across substances, gene mutation was predicted for 4-(phenylazo)benzene-1,3-diamine acetate (CAS no 79234 -33 -6). The study assumed the use of Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and TA 102 with and without S9 metabolic activation system. 4-(phenylazo)benzene-1,3-diamine acetate was predicted to induce gene mutation in Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and TA 102 in the presence and absence of S9 metabolic activation system and hence, according to the prediction made, it is likely to classify as a gene mutant in vitro.

The above mentioned predicted data is further supported by the data from read across chemicals as mentioned below:

In a study by Zeiger et al (Environmental Mutagenesis, 1987), Salmonella/microsome test in the absence of exogenous metabolic activation and in the presence of liver S-9 from Aroclor-induced male Sprague-Dawley rats and Syrian hamsters was performed to evaluate the mutagenic nature of 80 -90% struturally and functionally similar read across chemical Basic orange 2 (RA CAS no 532 -82 -1) using S. typhimurium tester strains TA1535, TA97, TA98 and TA100. The study was performed as per the preincubation assay and the preincubation time was 20 mins and the plates were incubated for 48 hrs. The test compound was dissolved in DMSO and was used at a dosage level of 0, 1.0, 3.0, 10, 16, 33, 100, 166, 333 or 1000 µg/plate in the preincubation assay of 48 hrs. Concurrent solvent and positive control chemicals were included in the study. Basic orange 2 induced a reproducible, dose-related increase in his⁺revertants over the corresponding solventin the S. typhimurium tester strains TA100, TA1537, and TA98 in the presence of S9 metabolic activation system. No mutagenic potential was observed for the tester strain TA1537 in the presence and absence of S9 metabolic activation system. Based on these observations, Basic orange 2 is likely to be mutagenic.

In another study by Sandhu and Chipman (Toxicology letters, 1991), Salmonella/S9 mutagenicity assay was performed to evaluate the mutagenic nature of another 80 -90% structurally and functionally similar read across chemical chrysoidine R (RA CAS no 495 -54 -5) with or without the addition of β-glucuronidase (βG). Chrysoidine R was dissolved in dimethyl sulphoxide (DMSO) and added (20 µL) at a concentration of 5-80 µg/plate. The test compound was found to increase the number of revertant colonies per plate and hence is considered to be mutagenic in vitro.

 

Based on the data available for the target chemical and read across, it can be concluded that the substance 4-(phenylazo)benzene-1,3-diamine acetate (CAS no 79234 -33 -6) exhibits gene mutation toxicity in vitro and hence it is likely to be classifed as mutagenic as per the criteria mentioned in CLP regulation.

Justification for classification or non-classification

Based on the data available for the target chemical and read across, it can be concluded that the substance 4-(phenylazo)benzene-1,3-diamine acetate (CAS no 79234 -33 -6) exhibits gene mutation toxicity in vitro and hence it is likely to be classifed as mutagenic as per the criteria mentioned in CLP regulation.