Disodium 2-[[5-carbamoyl-1-ethyl-1,6-dihydro-2-hydroxy-4-methyl-6-oxo-3-pyridyl]azo]-4-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]benzenesulphonate

Endpoint summary

Description of key information

The repeated dose toxicity study was predicted using OECD QSAR toolbox version 3.3 (2017) with log kow as the primary descriptor and considering the six closest read across substances; to evaluate the toxic effects of administration of disodium 2-[[5-carbamoyl-1 -ethyl-1,6 -dihydro-2-hydroxy-4- methyl-6- oxo- 3- pyridyl] azo]-4-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl] phenyl]amino]-1,3,5-triazin-2-yl] amino]ben zenesulphonate(CAS No. 84000-63-5) in rats by the oral route in a 13 weeks of exposure. No adverse effects were observed. Therefore, the no-observed adverse effect level (NOAEL) of disodium 2-[[5-carbamoyl -1-ethyl-1,6-dihydro-2-hydroxy-4-methyl-6-oxo-3-pyridyl] azo]-4-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]benzen esulphonate (CAS No. 84000-63-5) was estimated to be 629.28 mg/kg bw/day (nominal).

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

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:

no guideline followed

Principles of method if other than guideline:

Prediction is done using OECD QSAR Toolbox version 3.3 with log kow as the primary descriptor.

GLP compliance:

not specified

Specific details on test material used for the study:

- Name of test material: Disodium 2-[[5-carbamoyl-1-ethyl-1,6-dihydro-2-hydroxy-4-methyl -6-oxo-3-pyridyl]azo]- 4-[[4-chloro-6-[[3-[[2- (sulphona tooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]benzenesulphonate.
- Molecular formula : C26H24ClN9Na2O12S3
- Molecular weight : 832.1576 g/mol
- Smiles notation: CCn1c(c(c(c(c1=O)C (=O)N)C)/N=N/c2cc(ccc2S(=O)(=O)[O-]) Nc3nc(nc(n3)Cl)Nc4cccc(c4)S(=O)(=O)CCOS(=O) (=O)[O-])O. [Na+].[Na+]
- Substance type: Organic

Species:

rat

Strain:

Wistar

Sex:

male/female

Route of administration:

oral: gavage

Vehicle:

not specified

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

13 weeks

Frequency of treatment:

Daily

Dose / conc.:

629.28 mg/kg bw/day (nominal)

No. of animals per sex per dose:

10 male and 10 female rats

Control animals:

not specified

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Not specified
- Time schedule:Not specified
- Cage side observations checked in table [No.?] were included.: Not specified

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule:No data

BODY WEIGHT: Yes
- Time schedule for examinations:No data

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Not specified
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Not specified
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Not specified

FOOD EFFICIENCY: Not specified
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: Not specified

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Not specified
- Time schedule for examinations:Not specified

OPHTHALMOSCOPIC EXAMINATION: Not specified
- Time schedule for examinations:Not specified
- Dose groups that were examined:Not specified

HAEMATOLOGY: Not specified
- Time schedule for collection of blood:Not specified
- Anaesthetic used for blood collection: Not specified
- Animals fasted: Not specified
- How many animals:Not specified
- Parameters checked in table [No.?] were examined.Not specified

CLINICAL CHEMISTRY: Not specified
- Time schedule for collection of blood:Not specified
- Animals fasted:Not specified
- How many animals:Not specified
- Parameters checked in table [No.?] were examined.Not specified

URINALYSIS: Not specified
- Time schedule for collection of urine:Not specified
- Metabolism cages used for collection of urine: Not specified
- Animals fasted: Not specified
- Parameters checked in table [No.?] were examined.Not specified

NEUROBEHAVIOURAL EXAMINATION: Not specified
- Time schedule for examinations: Not specified
- Dose groups that were examined: Not specified
- Battery of functions tested: sensory activity / grip strength / motor activity / other: Not specified

IMMUNOLOGY: Not specified
- Time schedule for examinations:Not specified
- How many animals:Not specified
- Dose groups that were examined:Not specified
- Parameters checked in table [No.?] were examined.Not specified

Sacrifice and pathology:

GROSS PATHOLOGY: Yes

HISTOPATHOLOGY: Yes

Clinical signs:

no effects observed

Mortality:

not specified

Body weight and weight changes:

no effects observed

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:

no effects observed

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Dose descriptor:

NOAEL

Effect level:

629.28 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

clinical signs

gross pathology

histopathology: non-neoplastic

other: No effects observed.

Critical effects observed:

no

The prediction was based on dataset comprised from the following descriptors: NOAEL
Estimation method: Takes average value from the 6 nearest neighbours
Domain  logical expression:Result: In Domain

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

Domain logical expression index: "a"

Referential boundary: The target chemical should be classified as Imides (Acute toxicity) AND Substituted Triazines (Acute toxicity) AND Vinyl Sulfones by US-EPA New Chemical Categories

Domain logical expression index: "b"

Referential boundary: The target chemical should be classified as Imides (Acute toxicity) OR Substituted Triazines (Acute toxicity) OR Vinyl Sulfones by US-EPA New Chemical Categories ONLY

Domain logical expression index: "c"

Referential boundary: The target chemical should be classified as Non-specific AND Non-specific >> Incorporation into DNA/RNA, due to structural analogy with  nucleoside bases    AND Non-specific >> Incorporation into DNA/RNA, due to structural analogy with  nucleoside bases    >> Specific Imine and Thione Derivatives AND Radical AND Radical >> Radical mechanism via ROS formation (indirect) AND Radical >> Radical mechanism via ROS formation (indirect) >> Specific Imine and Thione Derivatives AND SN1 AND SN1 >> Nucleophilic substitution on diazonium ions AND SN1 >> Nucleophilic substitution on diazonium ions >> Specific Imine and Thione Derivatives by DNA binding by OASIS v.1.3

Domain logical expression index: "d"

Referential boundary: The target chemical should be classified as SN1 AND SN1 >> Iminium Ion Formation AND SN1 >> Iminium Ion Formation >> Aliphatic tertiary amines by DNA binding by OECD

Domain logical expression index: "e"

Referential boundary: The target chemical should be classified as Nucleophilic addition AND Nucleophilic addition >> Addition to carbon-hetero double bonds AND Nucleophilic addition >> Addition to carbon-hetero double bonds >> Ketones AND SNAr AND SNAr >> Nucleophilic aromatic substitution on activated aryl and heteroaryl compounds AND SNAr >> Nucleophilic aromatic substitution on activated aryl and heteroaryl compounds >> Activated aryl and heteroaryl compounds by Protein binding by OASIS v1.3

Domain logical expression index: "f"

Referential boundary: The target chemical should be classified as Acylation AND Acylation >> Direct Acylation Involving a Leaving group AND Acylation >> Direct Acylation Involving a Leaving group >> Acetates AND SNAr AND SNAr >> Nucleophilic aromatic substitution AND SNAr >> Nucleophilic aromatic substitution >> Halo-triazines by Protein binding by OECD

Domain logical expression index: "g"

Referential boundary: The target chemical should be classified as Acid moiety AND Acrylamides AND Hydrazines AND Imides AND Salt AND Triazines, Aromatic by Aquatic toxicity classification by ECOSAR

Domain logical expression index: "h"

Referential boundary: The target chemical should be classified as Acid moiety OR Acrylamides OR Hydrazines OR Imides OR Salt OR Triazines, Aromatic by Aquatic toxicity classification by ECOSAR ONLY

Domain logical expression index: "i"

Referential boundary: The target chemical should be classified as Nucleophilic addition OR Nucleophilic addition >> Addition to carbon-hetero double bonds OR Nucleophilic addition >> Addition to carbon-hetero double bonds >> Ketones OR SNAr OR SNAr >> Nucleophilic aromatic substitution on activated aryl and heteroaryl compounds OR SNAr >> Nucleophilic aromatic substitution on activated aryl and heteroaryl compounds >> Activated aryl and heteroaryl compounds by Protein binding by OASIS v1.3 ONLY

Domain logical expression index: "j"

Referential boundary: The target chemical should be classified as longer than months by Biodeg ultimate (Biowin 3) ONLY

Domain logical expression index: "k"

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

Domain logical expression index: "l"

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

Domain logical expression index: "m"

Similarity boundary:Target: CCN1C(=O)C(C(N)=O)=C(C)C(=NNc2cc(Nc3nc(Nc4cccc(S(=O)(=O)CCOS(=O)(=O)O{-}.[Na]{+})c4)nc(Cl)n3)ccc2S(=O)(=O)O{-}.[Na]{+})C1=O
Threshold=10%,
Dice(Atom centered fragments)
Atom type; Count H attached; Hybridization

Domain logical expression index: "n"

Similarity boundary:Target: CCN1C(=O)C(C(N)=O)=C(C)C(=NNc2cc(Nc3nc(Nc4cccc(S(=O)(=O)CCOS(=O)(=O)O{-}.[Na]{+})c4)nc(Cl)n3)ccc2S(=O)(=O)O{-}.[Na]{+})C1=O
Threshold=30%,
Dice(Atom centered fragments)
Atom type; Count H attached; Hybridization

Domain logical expression index: "o"

Parametric boundary:The target chemical should have a value of log Kow which is >= -5.64

Domain logical expression index: "p"

Parametric boundary:The target chemical should have a value of log Kow which is <= -0.104

Conclusions:

The no-observed adverse effect level (NOAEL) of disodium 2-[[5-carbamoyl-1-ethyl-1,6-dihydro-2-hydroxy-4-methyl-6-oxo-3-pyridyl] azo]-4-[[4-chlo ro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]benzenesulphonate (CAS No. 84000-63-5) was estimated to be 629.28 mg/kg bw/day (nominal).

Executive summary:

The repeated dose toxicity study was predicted using OECD QSAR toolbox version 3.3 (2017) with log kow as the primary descriptor and considering the six closest read across substances; to evaluate the toxic effects of administration of disodium 2-[[5-carbamoyl-1 -ethyl-1,6 -dihydro-2-hydroxy-4- methyl-6- oxo- 3- pyridyl] azo]-4-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl] phenyl]amino]-1,3,5-triazin-2-yl] amino]ben zenesulphonate(CAS No. 84000-63-5) in rats by the oral route in a 13 weeks of exposure. No adverse effects were observed. Therefore, the no-observed adverse effect level (NOAEL) of disodium 2-[[5-carbamoyl -1-ethyl-1,6-dihydro-2-hydroxy-4-methyl-6-oxo-3-pyridyl] azo]-4-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]benzen esulphonate (CAS No. 84000-63-5) was estimated to be 629.28 mg/kg bw/day (nominal).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

629.28 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

The data is Klimicsh 2 and from OECD QSAR toolbox version 3.3 (2017).

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: oral

Various repeated dose toxicity studies has been investigated to observe the adverse general toxicological effects occurring as a result of repeated daily dosing with, or exposure, to a substance for a specified period up to the expected lifespan of the test species. Often are the studies based on experiments and estimated data in rodents fordisodium 2-[[5-carbamoyl-1-ethyl-1,6-dihydro-2-hydroxy-4-methyl-6-oxo-3-pyridyl] azo]-4-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]benzenesulphonate (CAS No.- 84000-63-5) along with the study available on structurally similar read across substances Disodium 6-hydroxy-5-[(2-methoxy-4-sulphonato-m-tolyl)azo]naphthalene-2-sulphonate (FD & C RED NO. 40)(CAS No.25956-17-6) and disodium 3-[(2,4-dimethyl-5-sulphonatop henyl)azo]-4-hydroxynaphthalene-1-sulphonate (CAS No. 4548-53-2). The predicted data using the OECD QSAR toolbox has also been compared with the experimental data. The studies are summarized as below: 

 

The repeated dose toxicity study was predicted using OECD QSAR toolbox version 3.3 (2017) with log kow as the primary descriptor and considering the six closest read across substances; to evaluate the toxic effects of administration of disodium 2-[[5-carbamoyl-1 -ethyl-1,6 -dihydro-2-hydroxy-4- methyl-6- oxo- 3- pyridyl] azo]-4-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl] phenyl]amino]-1,3,5-triazin-2-yl] amino]ben zenesulphonate(CAS No. 84000-63-5) in rats by the oral route in a 13 weeks of exposure. No adverse effects were observed. Therefore, the no-observed adverse effect level (NOAEL) of disodium 2-[[5-carbamoyl -1-ethyl-1,6-dihydro-2-hydroxy-4-methyl-6-oxo-3-pyridyl] azo]-4-[[4-chloro-6- [[3-[[2- (sulph onatooxy)ethyl]sulphonyl ]phenyl]amino]-1,3,5-triazin-2-yl]amino]benzen esulphonate (CAS No. 84000-63-5) was estimated to be 629.28 mg/kg bw/day (nominal). 

 

In addition, in a Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening conducted by T. F. X. COLLINS et al. (Food and cosmatics toxicology, Vol. 18. pp 561 to 568, 1980), Osborne-Mendelfemale rats were treated wtih FD & C RED NO. 40 in the concentration of 0, 7.5, 15, 30, 100 and 200 mg/kg body weight /day in distilled water by oral gavage. No external signs of toxicity and animals were appeared healthy and behaved normally in treated groups as compared to control. No effects on weight gain and water consumption were observed in treated female rats as compared to control. No significant effect on numbers of corpora lutea, implantations, early and late deaths and viable foetuses per litter or on the percentage of preimplantation loss were observed in treated group as compared to control. Slightly increase in percentage of early resorptions were observed in 7.5 and 15 mg/kg bw/day but this response were appeared to be the result of sporadic occurrences. One litter was totally resorbed at 100 mg/kg bw/day. But, the percentage of females with more than one and with more than two resorptions showed no dose-related correlation as comapred to control. In addition, no effect on live or dead foetuses and mean foetal body weight and percentage of males and females per treatment group were observed as compared to control. Increased in number of runts were observed at 7.5, 15, 100 and 200 mg/kg bw/day treated dams, No compound-related external variations were observed in litters of treated dams as compared to control. Slightly decrease in males and females Crown-rump length were observed but were not significantly significant with increasing dose level. No effect on percentage of males and females per treatment group were observed as compared to control. Therefore, NOAEL was considered to be 200 mg/kg body weight /day when Osborne-Mendel female rats were treated wtih FD & C RED NO. 40 orally by gavage for 19 days. 

 

In a similar type of study conducted by T. F. X. COLLINS et al. (Food and cosmatics toxicology, Vol. 18. pp 561 to 568, 1980), Osborne-Mendel female rats were treated wtih FD & C RED NO. 40 in the concentration of 260.2 mg/kg body weight /day in distilled water by oral drinking water. No external signs of toxicity and animals were appeared healthy and behaved normally in treated groups as compared to control.No effects on maternal weight gain and Significant decrease in water consumption were observed in treated female rats as compared to control. No significant effect on numbers of corpora lutea, implantations, early and late deaths and viable foetuses per litter or on the percentage of preimplantation loss were observed in treated group as compared to control. Similarly, no significant effect on percentage of early resorptions and percentage of females with more than one and with more than two resorptions were observed in treated rats as compared to control. In addition, no effect on live or dead foetuses and mean foetal body weight and percentage of males and females per treatment group were observed as compared to control. No external variations were observed in litters of treated dams as compared to control. No effect on Crown-rump length and sex distribution were observed in litters of treated dams as compared to control.Slight increase in number of affected litters, number of bipartite sternebrae and reduced ossification of the parietal bone were observed in litters of treated dams as compared to control. But, the observed increases did not appear to be dose-related. Therefore,NOAEL was considered to be 260.2 mg/kg body weight /day when Osborne-Mendel female rats were treated wtih FD & C RED NO. 40. orally in Drinking water for 20 days. 

 

Moreover, the chronic repeated dose toxicity study was performed on Osborne-Mendel rats (Five groups of 24 litter mated) by K. J. DAVIS et al. (Toxicology And Applied Pharamacology 8, 306-317 (1966)) . Total 120 rats were used from which 16 were discarded because of advanced postmortem autolysis and 104 were received in the pathology laboratory for gross and microscopic examination.Six male and six female rats, evenly divided between control animals and those fed Disodium 3-[(2,4-dimethyl-5- sulp honatophenyl) azo]-4-hydroxynaphthalene-1-sulphonate (Ponceau SX) at 5%, were examined in detail. Of the12 animals, examinations were made of urinary bladder of 11 animals, prostate of 6, and parathyroid of 6. Liver, kidney, any tumors present, and testis in males were examined in all the remaining 92 rats. 

Gross lesions most commonly seen were tumors and granular kidneys but none was related to Ponceau SX treatment. No microscopic pathologic changes were attributed to Ponceau SX. No microscopic pathologic changes were attributed to Ponceau SX. No effect on survival, haematology and organ weight was observed. Therefore, the NOAEL was considered to be 5% (2500 mg/kg bw/day) when Osborne-Mendel female rats were treated wtih Disodium 3-[(2, 4-dim ethyl-5 -sulp honatophenyl)azo]-4-hydroxynaphthalene-1-sulphonate (Ponceau SX) (CAS No. 4548-53-2) orally.

 

On the basis of evidence from above studies (target substance and to its read across substances) in experimental animals, it can be presumed that there is no potential to be harmful to human health following repeated exposure. The substance disodium 2-[[5-carbamoyl-1-ethyl-1,6-dihydro-2-hydroxy-4-methyl-6-oxo-3-pyridyl] azo]-4-[[4-chloro-6-[[3- [[2-(sulphonatooxy)ethyl]sulphonyl] phenyl] amino]-1,3,5-triazin-2-yl]amino]benzenesulphonate (CAS No.- 84000-63-5) is unclassified because no specific target organ toxicity was seen at or below the dose/concentration mentioned. Thus, on the basis of CLP classification criteria the substance is not classified.

 

Repeated dose toxicity: inhalation 

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 particle size distribution of the substance the majority of the particles were found to be in the size of 106.0 micrometer which is much larger size range compared to the inhalable particulate matter. Thus, exposure to inhalable dust, mist and vapour of the chemicaldisodium 2-[[5-carbamoyl-1-ethyl-1,6-dihydro-2-hydroxy-4-methyl-6-oxo-3-pyridyl] azo]-4-[[4-chloro- 6-[[3-[[2- (sulpho natooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]benzene sulphonate is highly unlikely. Therefore this study is considered for waiver.

 

Repeated dose toxicity: dermal

The acute toxicity value for disodium 2-[[5-carbamoyl-1-ethyl-1,6-dihydro-2-hydroxy-4-methyl-6-oxo-3-pyridyl] azo]-4-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl] sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]benzenesulphonate(as provided in section 7.2.3) is >2000 mg/kg body weight. The substance was also found to be not irrtating to skin. Also, given the use of the chemical as reactive dye; repeated exposure by the dermal route is unlikely. Thus, it is expected that disodium 2-[[5-carb amoyl-1- ethyl- 1,6-dihydro-2-hydroxy-4-methyl-6-oxo-3-pyridyl] azo]-4-[[4-chloro-6-[[3-[[2-(sulphonatooxy) ethyl]sulphonyl] phenyl] amino]-1,3,5-triazin- 2-yl]amino]benzenesulphonateshall not exhibit 28 day repeated dose toxicity by the dermal route. In addition, there is no dermal absorption data as well as no data available that suggests that disodium 2-[[5-carbamoyl- 1-ethyl-1,6- dihydro-2-hyd roxy-4-methyl-6-oxo-3-pyridyl] azo]-4-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]amino]-1,3,5-triazin-2-yl]amino]benzene sulphonate shall exhibit repeated dose toxicity by the dermal route. Hence this end point was considered for waiver.

Justification for classification or non-classification

Based on the available data for the assessment of repeated dose toxicity by oral, inhalation and dermal route and following CLP Regulation (EC) No 1272/2008 , the substance disodium 2-[[5-carbamoyl-1-ethyl-1,6-dihydro-2-hydroxy-4-methyl-6-oxo-3-pyridyl] azo]-4-[[4-chloro-6-[[3-[[2-(sulphonatooxy)ethyl] sulphonyl]phenyl] amino] -1,3,5-triazin-2-yl]amino]benzenesulphonate is not classified.