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Description of key information

The repeated dose toxicity study was predicted using OECD QSAR toolbox version 3.4 (2017) with log kow as the primary descriptor and considering the four closest read across substances; to evaluate the toxic effects of administration of 7-[[2-[(aminocarbonyl)amino]-4-[(4-amino-6- chloro-1,3,5-triazin-2-yl)amino] phenyl]azo]naphthalene-1,3,6-trisulphonic acid (CAS No. 35642-64-9) 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 7-[[2-[(aminocarbonyl)amino]-4-[(4-amino-6-chloro-1,3,5-triazin-2-yl)amino]phenyl] azo]naphthalene- 1,3,6-trisulphonic acid (CAS No. 35642-64-9) was estimated to be 681.29 mg/kg bw/day (actual dose received).

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
sub-chronic 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 3.4 and the supporting QMRF report has been attached.
Reference:
Composition 0
Qualifier:
no guideline available
Principles of method if other than guideline:
Prediction is done using OECD QSAR Toolbox version 3.4 with respect to the descriptor log Kow.
GLP compliance:
not specified
Test material information:
Composition 1
Specific details on test material used for the study:
Name of the test chemical: 7-[[2-[(aminocarbonyl)amino]-4-[(4-amino-6-chloro-1,3,5-triazin-2-yl)amino]phenyl]azo]naphthalene-1,3,6-trisulphonic acid
Molecular formula: C20H16ClN9O10S3
Molecular weight: 674.0504 g/mol
Smiles Notation: S(=O)(=O)(c1c(/N=N/c2c(NC(=O)N)cc(Nc3nc(nc(n3)Cl)N)cc2)cc2c(S(=O)(=O)O)cc(S(=O)(=O)O)cc2c1)O
InChI: 1S/C20H16ClN9O10S3/c21-17-26-18(22)28-20(27-17)24-9-1-2-12(13(5-9)25-19(23)31)29-30-14-7-11-8(4-16(14)43(38,39)40) 3-10(41(32,33)34)6-15(11)42(35,36)37/h1-7H,(H3,23,25,31)(H,32,33,34)(H,35,36,37)(H,38,39,40)(H3,22,24,26,27,28)/b30-29+
Substance Type: Organic
Physical State: solid
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.:
681.29 mg/kg bw/day (actual dose received)
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: Yes
- Time schedule:Not specified
- Cage side observations checked in table [No.?] were included.: Mortality was observed.

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):
- 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:
- 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:
no mortality observed
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:
681.29 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
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 5 nearest neighbours
Domain  logical expression:Result: In Domain

(((("a" or "b" )  and "c" )  and "d" )  and ("e" and "f" )  )

Domain logical expression index: "a"

Referential boundary: The target chemical should be classified as Acid moiety OR Anilines (Unhindered) OR Substituted Ureas OR Triazines, Aromatic by Aquatic toxicity classification by ECOSAR ONLY

Domain logical expression index: "b"

Referential boundary: The target chemical should be classified as Activated N-heterocycles by Skin irritation/corrosion Inclusion rules by BfR

Domain logical expression index: "c"

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: "d"

Referential boundary: The target chemical should be classified as Not bioavailable by Lipinski Rule Oasis ONLY

Domain logical expression index: "e"

Parametric boundary:The target chemical should have a value of log Kow which is >= -1.93

Domain logical expression index: "f"

Parametric boundary:The target chemical should have a value of log Kow which is <= 6.5

Conclusions:
The no-observed adverse effect level (NOAEL) of 7-[[2-[(aminocarbonyl)amino]-4-[(4-amino-6-chloro-1,3,5-triazin-2-yl)amino]phenyl]azo]naphthalene- 1,3,6-trisulphonic acid (CAS No. 35642-64-9) was estimated to be 681.29 mg/kg bw/day (actual dose received).

Executive summary:

The repeated dose toxicity study was predicted using OECD QSAR toolbox version 3.4 (2017) with log kow as the primary descriptor and considering the four closest read across substances; to evaluate the toxic effects of administration of 7-[[2-[(aminocarbonyl)amino]-4-[(4-amino-6- chloro-1,3,5-triazin-2-yl)amino] phenyl]azo]naphthalene-1,3,6-trisulphonic acid (CAS No. 35642-64-9) 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 7-[[2-[(aminocarbonyl)amino]-4-[(4-amino-6-chloro-1,3,5-triazin-2-yl)amino]phenyl] azo]naphthalene- 1,3,6-trisulphonic acid (CAS No. 35642-64-9) was estimated to be 681.29 mg/kg bw/day (actual dose received).

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
681.29 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
The data is Klimicsh 2 and from OECD QSAR toolbox version 3.4 (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

Mode of Action Analysis / Human Relevance Framework

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 for 7-[[2-[(aminocarbonyl)amino]-4-[(4-amino-6-chloro-1,3,5-triazin-2-yl)amino]phenyl]azo]naphthalene-1,3, 6-trisulphonic acid along with the study available on structurally similar read across substances Red 2G (CAS No.- 3734-67-6), disodium 3-[(2,4-dimethyl-5- sulp honatophenyl)azo]-4-hydroxynaphthalene-1-sulphonate (CAS No. 4548-53-2) and FD & C RED NO. 40 (CAS No. 25956-17-6). 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 3.4 (2017) with log kow as the primary descriptor and considering the four closest read across substances; to evaluate the toxic effects of administration of 7-[[2-[(aminocarbonyl)amino]-4-[(4-amino-6- chloro-1,3,5-triazin-2-yl)amino] phenyl]azo]naphthalene-1,3,6-trisulphonic acid (CAS No. 35642-64-9) 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 7-[[2-[(aminocarbonyl)amino]-4-[(4-amino-6-chloro-1,3,5-triazin-2-yl)amino]phenyl] azo]naph thalene- 1,3,6-trisulphonic acid (CAS No. 35642-64-9) was estimated to be 681.29 mg/kg bw/day (actual dose received).

 

Also, the subchronic repeated dose toxicity study on structurally similar read across substance Red 2G (CAS No.- 3734-67-6) in mice was published in an European Food Safety AuthorityJournal (The EFSA Journal (2007) 515, 1-28). Five groups of 15 male and 15 female mice were given diets containing 0, 0.01, 0.1, 1, or 2% Red 2G (equivalent to 14, 143, 1429, or 2858 mg/kg bw) for three months. 5 animals per group were haematologically investigated and fully autopsied at days 26, 55, and 96. A dose- dependent increase in the incidence of Heinz bodies was observed. Splenomegaly was observed in both sexes at 2% and in females at 1%. Relative liver weig hts were increased in females at 1% and 2% at day 26, and at 2% at days 55 and 96. Haemosiderin was increased in the liver (Kupffer cells) at 1% and 2%. In the spleen haemosiderin was increased at 2% at all test days, and at the other concentrations increased with treatment and duration. No adverse effects on growth or food consumption were evident. Thus under the condition of the study, the no-observed adverse effect level (NOAEL) of Red 2G (CAS No.- 3734-67-6) in this subchronic study in mice was observed to be 143.0 mg/kg bw. 

 

In addition to these studies, the chronic repeated dose toxicity study was performed by K. J. Davis et al., (Toxicology And Applied Pharamacology 8, 306-317 (1966)) on Osborne-Mendel rats (Five groups of 24 litter mated). 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 the 12 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. 

 

Moreover, in the study 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.

 

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 7-[[2-[(aminocarbonyl)amino]-4-[(4-amino-6-chloro-1,3,5-triazin -2-yl) amino]phenyl]azo]naphthalene-1,3,6-trisulphonic acid (CAS No. 35642-64-9) is unclassified because no specific target organ toxicity was seen. 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 low vapour pressure of the substance 7-[[2-[(aminocarbonyl)amino]-4-[(4-amino-6-chloro-1,3,5-triazin-2-yl)amino]phenyl]azo]napht halene- 1,3,6 -trisulphonic acid, which is reported as 3.72E-033 Pa. Also considering the particle size distribution of the substance the majority of the particles were found to be in the size of 103.0 micrometer which is much larger size range compared to the inhalable particulate matter. Thus, exposure to inhalable dust, mist and vapour of the chemical 7-[[2-[(aminocarbonyl)amino]-4-[(4-amino-6-chloro-1,3,5-triazin-2-yl)amino]phenyl]azo]naphthalene-1,3,6-trisulphonic acid is highly unlikely. Therefore this study is considered for waiver.

 

Repeated dose toxicity: dermal

The acute toxicity value for 7-[[2-[(aminocarbonyl)amino]-4-[(4-amino-6-chloro-1,3,5-triazin-2-yl)amino]phenyl]azo]naphthalene-1,3,6-trisulphonic acid (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 7-[[2-[(aminocarbonyl)amino]-4-[(4-amino-6-chloro-1,3,5- triazin-2-yl) amino]phenyl]azo]naphthalene-1,3,6-trisulphonic acid shall 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 7-[[2-[(aminocarbo nyl)amino]-4-[(4-amino-6-chloro-1,3,5-triazin-2-yl)amino]phenyl]azo] naphthalene-1,3,6 -trisulphonic acid 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 7-[[2-[(aminocarbonyl)amino]-4-[(4-amino-6-chloro-1,3,5-triazin-2-yl)amino]phenyl]azo]naphthalene-1,3,6-trisulphonic acid is not classified.