Disodium [[4-[bis[4-[(sulphonatophenyl)amino]phenyl]methylene]cyclohexa-2,5-dien-1-ylidene]amino]benzenesulphonate

Endpoint summary

Description of key information

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, repeated dose oral toxicity was predicted for the test compound Disodium [[4-[bis[4-[(sulphonatophenyl) amino]phenyl] methylene]cyclohexa-2,5-dien-1-ylidene]amino]ben..(28983-56-4 ). The study assumed the use of male and female Wistar strain in Subchronic study. No significant alterations were noted at the dose level of 815.90mg/Kg bw/day. The predicted No Observed Adverse Effect Level (NOAEL) for compound Disodium [[4-[bis[4-[(sulphonatophenyl) amino]phenyl] methylene]cyclohexa-2,5-dien-1-ylidene]amino]ben..is considered to be 815.90mg/Kg bw/day. Based on this value it can be concluded that the substance is considered to not toxic as per the criteria mentioned in CLP regulation.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

repeated dose toxicity: oral, other

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:

Prediction is done using OECD QSAR Toolbox version 3.3 and the supporting QMRF report has been attached.

Qualifier:

according to guideline

Guideline:

other: As mention below

Principles of method if other than guideline:

Prediction is done using OECD QSAR Toolbox version 3.3, 2018.

GLP compliance:

not specified

Limit test:

no

Specific details on test material used for the study:

- Name of test material :Methyl Blue
- Molecular formula : C37H26N3Na2O9S3
- Molecular weight : 800.8182 g/mol
- Smiles notation : c1cc(ccc1C(=C2C=CC(=[NH+]c3ccc (cc3)S(=O) (=O)[O-])C=C2)c4ccc(cc4)Nc5ccc(cc5)S (=O)(=O)[O-])Nc6ccc(cc6)S(=O)(=O)O.[Na+].[Na+]
- InChl : 1S/C37H29N3O9S3.2Na/c41-50(42,43)34-19-13-31(14-20-34)38-28-7-1-25(2-8-28)37(26-3-9-29(10-4-26)39-32-15-21-35(22-16-32)51(44,45 46)27-5-11-30(12-6-27)40-33-17-23-36(24-18-33)52(47,48)49;;/h1-24,38-39H,(H,41,42,43)(H,44,45,46)(H,47,48,49);;/q;2*+1/p-1
- Substance type: Organic
- Physical state: Solid

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

Not specified.

Route of administration:

oral: gavage

Details on route of administration:

Not specified.

Vehicle:

not specified

Details on oral exposure:

Not specified.

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

Not specified.

Duration of treatment / exposure:

80 days

Frequency of treatment:

Not specified.

Remarks:

Not specified.

No. of animals per sex per dose:

Not specified.

Control animals:

not specified

Details on study design:

Not specified.

Positive control:

Not specified.

Observations and examinations performed and frequency:

Not specified.

Sacrifice and pathology:

Not specified.

Other examinations:

Not specified.

Statistics:

Not specified.

Clinical signs:

not specified

Mortality:

not specified

Body weight and weight changes:

not specified

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

not specified

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

not specified

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Dose descriptor:

NOAEL

Effect level:

815.9 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No significant effect were observed at this dose.

Critical effects observed:

not specified

Treatment related:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

The prediction was based on dataset comprised from the following descriptors: NOAEL
Estimation method: Takes average value from the 5 nearest neighbours
Domain  logical expression:Result: In Domain

(((((((((((("a" or "b" or "c" or "d" )  and ("e" and ( not "f") )  )  and ("g" and ( not "h") )  )  and "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 "u" )  and ("v" and "w" )  )

Domain logical expression index: "a"

Referential boundary: The target chemical should be classified as Alkene OR Ammonium salt OR Aromatic amine OR Aryl OR Sulfonic acid by Organic Functional groups ONLY

Domain logical expression index: "b"

Referential boundary: The target chemical should be classified as Alkene OR Ammonium salt OR Aromatic amine OR Aryl OR Overlapping groups OR Sulfonic acid by Organic Functional groups (nested) ONLY

Domain logical expression index: "c"

Referential boundary: The target chemical should be classified as Aliphatic Nitrogen, two aromatic attach [-N-] OR Aromatic Carbon [C] OR Miscellaneous sulfide (=S) or oxide (=O) OR Nitrogen, hydrogen attach {v+5} OR Nitrogen, two or tree olefinic attach [>N-] OR Olefinic carbon [=CH- or =C<] OR Ortho-substitutes on N=C<, aromatic OR Suflur {v+4} or {v+6} OR Sulfonate, aromatic attach [-SO2-O] by Organic functional groups (US EPA) ONLY

Domain logical expression index: "d"

Referential boundary: The target chemical should be classified as Amine OR Anion OR Aromatic compound OR Cation OR Secondary amine OR Secondary aromatic amine OR Sulfonic acid derivative by Organic functional groups, Norbert Haider (checkmol) ONLY

Domain logical expression index: "e"

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

Domain logical expression index: "f"

Referential boundary: The target chemical should be classified as Michael addition OR Michael addition >> P450 Mediated Activation to Quinones and Quinone-type Chemicals OR Michael addition >> P450 Mediated Activation to Quinones and Quinone-type Chemicals >> Arenes OR Michael addition >> P450 Mediated Activation to Quinones and Quinone-type Chemicals >> Polycyclic (PAHs) and heterocyclic (HACs) aromatic hydrocarbons-Michael addition OR Michael addition >> Polarised Alkenes-Michael addition OR Michael addition >> Polarised Alkenes-Michael addition >> Alpha, beta- unsaturated ketones OR SN1 OR SN1 >> Carbenium Ion Formation OR SN1 >> Carbenium Ion Formation >> Polycyclic (PAHs) and heterocyclic (HACs) aromatic hydrocarbons-SN1 OR SN1 >> Nitrenium Ion formation OR SN1 >> Nitrenium Ion formation >> Tertiary aromatic amine by DNA binding by OECD

Domain logical expression index: "g"

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

Domain logical expression index: "h"

Referential boundary: The target chemical should be classified as AN2 OR AN2 >> Nucleophilic addition to alpha, beta-unsaturated carbonyl compounds OR AN2 >> Nucleophilic addition to alpha, beta-unsaturated carbonyl compounds >> alpha, beta-Unsaturated Aldehydes OR AN2 >> Schiff base formation OR AN2 >> Schiff base formation >> alpha, beta-Unsaturated Aldehydes 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 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 by DNA binding by OASIS v.1.3

Domain logical expression index: "i"

Referential boundary: The target chemical should be classified as Class 5 (Not possible to classify according to these rules) by Acute aquatic toxicity classification by Verhaar (Modified) ONLY

Domain logical expression index: "j"

Referential boundary: The target chemical should be classified as Inclusion rules not met by Skin irritation/corrosion Inclusion rules by BfR

Domain logical expression index: "k"

Referential boundary: The target chemical should be classified as Quaternary organic ammonium compounds by Skin irritation/corrosion Inclusion rules by BfR

Domain logical expression index: "l"

Referential boundary: The target chemical should be classified as (!Undefined)Group All Lipid Solubility < 0.01 g/kg AND (!Undefined)Group CNS Surface Tension > 62 mN/m AND Group All log Kow < -3.1 AND Group All Melting Point > 200 C AND Group CNS log Kow < 0.5 AND Group CNS log Kow < -2 AND Group CNS Melting Point > 120 C AND Group CNS Melting Point > 50 C AND Group CNS Molecular Weight > 620 g/mol by Skin irritation/corrosion Exclusion rules by BfR

Domain logical expression index: "m"

Referential boundary: The target chemical should be classified as Group All log Kow > 9 by Skin irritation/corrosion Exclusion rules by BfR

Domain logical expression index: "n"

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

Domain logical expression index: "o"

Referential boundary: The target chemical should be classified as Halogens by Groups of elements

Domain logical expression index: "p"

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

Domain logical expression index: "q"

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

Domain logical expression index: "r"

Referential boundary: The target chemical should be classified as Not categorized by Repeated dose (HESS)

Domain logical expression index: "s"

Referential boundary: The target chemical should be classified as Aliphatic nitriles (Hepatotoxicity) Rank B OR Benzene/ Naphthalene sulfonic acids (Less susceptible) Rank C OR Tamoxifen (Hepatotoxicity) Alert by Repeated dose (HESS)

Domain logical expression index: "t"

Referential boundary: The target chemical should be classified as Alkene AND Ammonium salt AND Aromatic amine AND Aryl AND Sulfonic acid by Organic Functional groups ONLY

Domain logical expression index: "u"

Referential boundary: The target chemical should be classified as Alkene AND Ammonium salt AND Aromatic amine AND Aryl AND Sulfonic acid by Organic Functional groups ONLY

Domain logical expression index: "v"

Parametric boundary:The target chemical should have a value of log Kow which is >= -4.46

Domain logical expression index: "w"

Parametric boundary:The target chemical should have a value of log Kow which is <= -3.4

Conclusions:

The predicted No Observed Adverse Effect Level (NOAEL) for Disodium [[4-[bis[4-[(sulphonatophenyl)amino]phenyl]methylene]cyclohexa-2,5-dien-1-ylidene]amino]ben..(28983-56-4 )is considered to be 815.90mg/Kg bw/day.

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, repeated dose oral toxicity was predicted for the test compound Disodium [[4-[bis[4-[(sulphonatophenyl) amino]phenyl] methylene]cyclohexa-2,5-dien-1-ylidene]amino]ben..(28983-56-4 ). The study assumed the use of male and female Wistar strain in Subchronic study. No significant alterations were noted at the dose level of 815.90mg/Kg bw/day. The predicted No Observed Adverse Effect Level (NOAEL) for compound Disodium [[4-[bis[4-[(sulphonatophenyl) amino]phenyl] methylene]cyclohexa-2,5-dien-1-ylidene]amino]ben..is considered to be 815.90mg/Kg bw/day. Based on this value it can be concluded that the substance is considered to not toxic as per the criteria mentioned in CLP regulation.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

815.9 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

K2 data from prediction OECD QSAR toolbox 3.3

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Repeated dose toxicity: via oral route;

Various experimental studies were reviewed to determine the toxic nature of target substance Methyl Blue; IUPAC;Disodium [[4-[bis[4-[(sulphonatophenyl) amino]phenyl] methylene]cyclohexa-2,5-dien-1-ylidene]amino]ben..(28983-56-4 ) upon repeated exposure by oral route. The studies are as mentioned 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, repeated dose oral toxicity was predicted for the test compound Disodium [[4-[bis[4-[(sulphonatophenyl) amino]phenyl] methylene]cyclohexa-2,5-dien-1-ylidene]amino]ben..(28983-56-4 ). The study assumed the use of male and female Wistar strain in Subchronic study. No significant alterations were noted at the dose level of 815.90mg/Kg bw/day. The predicted No Observed Adverse Effect Level (NOAEL) for compound Disodium [[4-[bis[4-[(sulphonatophenyl) amino]phenyl] methylene]cyclohexa-2,5-dien-1-ylidene]amino]ben..is considered to be 815.90mg/Kg bw/day. Based on this value it can be concluded that the substance is considered to not toxic as per the criteria mentioned in CLP regulation.

Another  Sub-chronic toxicity study for Read across was performed by S. A. Clode et al.( Fd Chem. Toxic., 1987) to determine the oral toxic nature of Amaranth; IUPAC trisodium 3-hydroxy-4-[(4-sulfonato-1-naphthyl)diazenyl]naphthalene-2,7-disulfonate (915-67-3). The read across share high similarity in structure and log kow .Therefore, it is acceptable to derive information on mutation from the analogue substance. In acombined repeated dose & carcinogenicity,Wistar male and female rats were treated withAmaranth dye in the concentration of 47, 242 and 1260 mg/kg bw/day for male and 49, 246 and 1260 mg/kg bw/day for female orally in fed.No significant effect on survival of treated rats were observed as compared to control.Significant decrease in body weight in male rats from week 4 to 73 and in female rat from 51 and 73 and increase in food consumption was observed in 1260 mg/kg bw/day treated rats as compared to control. Slight effect on body weight and food consumption was due to a reduction in the absorption or utilization of the nutrient. Average compound intake of male rats were 47, 242 and 1260 mg/kg bw/day and for female rats 49, 246 and 1260 mg/kg bw/day.Increase in water consumption was observed in 1260 mg/kg bw/day treated male rats as compared to control. Increased water loss was observed due to production of softer, moister faeces and a compensatory increase in water intake. Similarly, decreased in packed cell volume was observed at month 6 and 12 in male and at month 18 in female rats and slightly increased haemoglobin concentrations in female rats at termination of study and decrease in glutamic-oxalacetic transaminase activities in male and female rats but not significant in female rats were observed at 1260 mg/kg bw/day. Observed effect were not dose related or consistent between the sexes. Increase level of proteinin urine at 12 months in female was observed at 1260 mg/kg bw/day andSemi-quantitative analysis of urine for bilirubin and ketones was hindered at months 18 and 24 due to the contamination of the urine with amaranth which interfered with the colour reaction. Statistically significant increase in absolute and relative full and empty caecum weight were observed in male and female at 1260mg/kg bw/day andincrease in absolute and relative full caecum weight in male rats at 242mg/kg bw/day were observedas compared to control. The observed effect was not statistically significant as compared to control. In addition, Non-neoplastic transitional-cell hyperplasia of bladder, inflammatory cell infiltrate of the seminal vesicles and testicular interstitial-cell hyperplasia were observed in 47 mg/kg bw/day male and 1260 mg/kg bw/day treated male and female rats. Statistically significant increase in renal calcification and renal pelvic epithelial hyperplasia, lung oedema and haemorrhage, lymph-node haemorrhage and degenerative changes in the brain and nerve, degenerative and inflammatory changes in heart, inflammatory changes in thymus, aortic calcification and atrial thrombi were observed in 1260 mg/kg bw/day treated female rats. Statistically significant increase in neoplastic uterine polyps and vaginal fibromas were observed in 1260 mg/kg bw/day treated female rats. The observed effects were typical of this strain and occur in ageing rats. No effect on number of litters, number of pups per litter at day 18 and pup weight at day 18 were observed in treated rats as compared to control. Therefore, NOAEL was considered to be 1260 mg/kg body weight /day when Wistar male and female rats were treated with Amaranth dye orally in fed for 2 years.

Repeated inhalation study:

According to Annex IX of the REACH regulation, testing by the inhalation route is appropriate only if exposure of humans via inhalation is likely. Taking into account the low vapour pressure of the substance Methyl Blue; IUPAC;Disodium [[4-[bis[4-[(sulphonatophenyl) amino]phenyl] methylene]cyclohexa-2,5-dien-1-ylidene]amino]ben..(28983-56-4 ) ,which is reported as 7.8526E-42 Pa at 25 C. Also considering the particle size distribution of the substance the majority of the particles was found to be in the size of 150 micron to 25 micron which is much larger size range compared to the inhalable particulate matter .Thus, exposure to inhalable dust, mist and vapour of the chemical Methyl Blue; IUPAC;Disodium [[4-[bis[4-[(sulphonatophenyl) amino]phenyl] methylene]cyclohexa-2,5-dien-1-ylidene]amino]ben..(28983-56-4 )  is highly unlikely. Therefore this study is considered for waiver.

 

Repeated dermal study;

The acute toxicity value for Methyl Blue; IUPAC;Disodium [[4-[bis[4-[(sulphonatophenyl) amino]phenyl] methylene]cyclohexa-2,5-dien-1-ylidene]amino]ben..(28983-56-4 ) (as provided in section 7.2.3) is 7167mg/kg body weight. Also, given the use of the chemical; repeated exposure by the dermal route is unlikely since the use of gloves is common practice in industries. Thus, it is expected that Methyl Blue; IUPAC;Disodium [[4-[bis[4-[(sulphonatophenyl) amino]phenyl] methylene]cyclohexa-2,5-dien-1-ylidene]amino]ben.shall not exhibit 28 day repeated dose toxicity by the dermal route. In addition, there is no data available that suggests that Methyl Blue; IUPAC;Disodium [[4-[bis[4-[(sulphonatophenyl) amino]phenyl] methylene]cyclohexa-2,5-dien-1-ylidene]amino]ben.. shall exhibit repeated dose toxicity by the dermal route. Hence this end point was considered for waiver.

 

Based on the data available for the target chemical and its prediction, Methyl Blue; IUPAC;Disodium [[4-[bis[4-[(sulphonatophenyl) amino]phenyl] methylene]cyclohexa-2,5-dien-1-ylidene]amino]ben..(28983-56-4 )does not exhibit toxic nature upon repeated exposure by oral, inhalation and dermal route of exposure and hence is not likely to classify as per the criteria mentioned in CLP regulation.

Justification for classification or non-classification

Based on the data available for the target chemical and its prediction, Methyl Blue; IUPAC;Disodium [[4-[bis[4-[(sulphonatophenyl) amino]phenyl] methylene]cyclohexa-2,5-dien-1-ylidene]amino]ben..(28983-56-4 )does not exhibit toxic nature upon repeated exposure by oral, inhalation and dermal route of exposure and hence is not likely to classify as per the criteria mentioned in CLP regulation.