N-phenyl-4-[[4-(phenylamino)phenyl][4-(phenylimino)cyclohexa-2,5-dien-1-ylidene]methyl]aniline monohydrochloride

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 N-phenyl-4-[[4-(phenylamino) phenyl][4-(phenylimino)cyclohexa-2 ,5-dien-1-ylidene] methyl]aniline monohydrochloride. The study assumed the use of male and female Sprague-Dawley rats in subchronic study of 5 weeks. No significant alterations were noted at the dose level of 850.04mg/Kg bw/day. The predicted No Observed Adverse Effect Level (NOAEL) for N-phenyl-4-[[4-(phenylamino)phenyl][4 -(phenylimino)cyclohexa-2,5-dien-1-ylidene]methyl]aniline monohydrochloride is considered to be 850.04mg/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

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, 2017

GLP compliance:

not specified

Limit test:

no

Specific details on test material used for the study:

- Name of test material: N-phenyl-4-[[4-(phenylamino)phenyl][4-(phenylimino)cyclohexa-2,5-dien-1-ylidene]methyl]aniline monohydrochloride
- Molecular formula: C37H29N3.ClH
- Molecular weight: 552.118 g/mol
- Smiles notation: C(\c1ccc(Nc2ccccc2)cc1)(c1ccc(Nc2ccccc2)cc1)=C1/C=C\C(=N/c2ccccc2)C=C1.Cl
- InChl: 1S/C37H29N3.ClH/c1-4-10-31(11-5-1)38-34-22-16-28(17-23-34)37(29-18-24-35(25-19-29)39-32-12-6-2-7-13-32)30-20-26-36(27-21-30)40-33-14-8-3-9-15-33;/h1-27,38-39H;1H
- Substance type: Organic
- Physical state: solid

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

No data available.

Route of administration:

oral: gavage

Details on route of administration:

No data available.

Vehicle:

not specified

Details on oral exposure:

No data available.

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

No data available.

Duration of treatment / exposure:

5 weeks

Frequency of treatment:

No data available.

Remarks:

No data available.

No. of animals per sex per dose:

No data available.

Control animals:

not specified

Details on study design:

No data available.

Positive control:

No data available.

Observations and examinations performed and frequency:

No data available.

Sacrifice and pathology:

No data available.

Other examinations:

No data available.

Statistics:

No data available.

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

Details on results:

No data available.

Dose descriptor:

NOAEL

Effect level:

850.045 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No treatment related effect were observed

Remarks on result:

other: No toxic effect were observed

Critical effects observed:

not specified

The prediction was based on dataset comprised from the following descriptors: NOAEL
Estimation method: Takes average value from the 10 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 ( not "j") )  )  and "k" )  and ("l" and ( not "m") )  )  and "n" )  and ("o" and ( not "p") )  )  and "q" )  and ("r" and "s" )  )

Domain logical expression index: "a"

Referential boundary: The target chemical should be classified as Alkene OR Aromatic amine OR Aryl OR Azomethine OR Ketimine OR No functional group found by Organic Functional groups ONLY

Domain logical expression index: "b"

Referential boundary: The target chemical should be classified as Alkene OR Aromatic amine OR Aryl OR Azomethine OR Ketimine OR No functional group found OR Overlapping groups by Organic Functional groups (nested) ONLY

Domain logical expression index: "c"

Referential boundary: The target chemical should be classified as Aliphatic Nitrogen, one aromatic attach [-N] OR Aliphatic Nitrogen, two aromatic attach [-N-] OR Aromatic Carbon [C] OR Nitrogen, two or tree olefinic attach [>N-] OR No functional group found OR Olefinic carbon [=CH- or =C<] by Organic functional groups (US EPA) ONLY

Domain logical expression index: "d"

Referential boundary: The target chemical should be classified as Amine OR Aromatic compound OR No functional group found OR Secondary amine OR Secondary aromatic amine 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 AND 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: "f"

Referential boundary: The target chemical should be classified as AN2 OR AN2 >> Carbamoylation after isocyanate formation OR AN2 >> Carbamoylation after isocyanate formation >> N-Hydroxylamines 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 Michael addition OR Michael addition >> Quinone type compounds OR Michael addition >> Quinone type compounds >> Quinone methides OR Radical >> Radical mechanism via ROS formation (indirect) >> Hydrazine Derivatives OR Radical >> Radical mechanism via ROS formation (indirect) >> N-Hydroxylamines OR Radical >> ROS formation after GSH depletion OR Radical >> ROS formation after GSH depletion >> Quinone methides OR SN1 >> Nucleophilic attack after metabolic nitrenium ion formation OR SN1 >> Nucleophilic attack after metabolic nitrenium ion formation >> N-Hydroxylamines by DNA binding by OASIS v.1.3

Domain logical expression index: "g"

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

Domain logical expression index: "h"

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 >> Polarised Alkenes-Michael addition OR Michael addition >> Polarised Alkenes-Michael addition >> Alpha, beta- unsaturated ketones by DNA binding by OECD

Domain logical expression index: "i"

Referential boundary: The target chemical should be classified as Non binder, MW>500 AND Non binder, non cyclic structure by Estrogen Receptor Binding

Domain logical expression index: "j"

Referential boundary: The target chemical should be classified as Non binder, impaired OH or NH2 group OR Non binder, without OH or NH2 group by Estrogen Receptor Binding

Domain logical expression index: "k"

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: "l"

Referential boundary: The target chemical should be classified as No alert found by Carcinogenicity (genotox and nongenotox) alerts by ISS

Domain logical expression index: "m"

Referential boundary: The target chemical should be classified as Azide and triazene groups (Genotox) OR Metals, oxidative stress (Nongenotox) OR Structural alert for genotoxic carcinogenicity OR Structural alert for nongenotoxic carcinogenicity by Carcinogenicity (genotox and nongenotox) alerts by ISS

Domain logical expression index: "n"

Referential boundary: The target chemical should be classified as Alkene AND Aromatic amine AND Aryl AND Azomethine AND Ketimine AND No functional group found by Organic Functional groups ONLY

Domain logical expression index: "o"

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

Domain logical expression index: "p"

Referential boundary: The target chemical should be classified as Aliphatic nitriles (Hepatotoxicity) Rank B OR Perhexiline (Hepatotoxicity) Alert by Repeated dose (HESS)

Domain logical expression index: "q"

Referential boundary: The target chemical should be classified as Amine AND Aromatic compound AND No functional group found AND Secondary amine AND Secondary aromatic amine by Organic functional groups, Norbert Haider (checkmol) ONLY

Domain logical expression index: "r"

Parametric boundary:The target chemical should have a value of log Kow which is >= 0.22

Domain logical expression index: "s"

Parametric boundary:The target chemical should have a value of log Kow which is <= 10.2

Conclusions:

The predicted No Observed Adverse Effect Level (NOAEL) for N-phenyl-4-[[4-(phenylamino)phenyl][4-(phenylimino)cyclohexa-2,5-dien-1-ylidene]methyl]aniline monohydrochloride is considered to be 850.04 mg/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 N-phenyl-4-[[4-(phenylamino)phenyl][4-(phenylimino)cyclohexa-2,5-dien-1-ylidene]methyl]aniline monohydrochloride. The study assumed the use of male and female Sprague-Dawley rats in subchronic study of 5 weeks. No significant alterations were noted at the dose level of 850.04mg/Kg bw/day. The predicted No Observed Adverse Effect Level (NOAEL) for N-phenyl-4-[[4- (phenylamino)phenyl][4- (phenylimino)cyclohexa- 2, 5-dien-1-ylidene]methyl]aniline monohydrochloride is considered to be 850.04mg/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

850.04 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

Data is ferom QSAR K2

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 oral toxicity:

Prediction model based estimation and data available for the target chemical was reviewed to determine the toxic nature of N-phenyl-4-[[4-(phenylamino)phenyl][4-(phenylimino)cyclohexa-2,5-dien-1-ylidene]methyl]aniline monohydrochloride (2152-64-9)upon repeated exposure by oral, dermal and inhalation route of exposure. 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 N-phenyl-4-[[4-(phenylamino) phenyl][4-(phenylimino)cyclohexa-2 ,5-dien-1-ylidene] methyl]aniline monohydrochloride. The study assumed the use of male and female Sprague-Dawley rats in subchronic study of 5 weeks. No significant alterations were noted at the dose level of 850.04mg/Kg bw/day. The predicted No Observed Adverse Effect Level (NOAEL) for N-phenyl-4-[[4-(phenylamino)phenyl][4 -(phenylimino)cyclohexa-2,5-dien-1-ylidene]methyl]aniline monohydrochloride is considered to be 850.04mg/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 Repeated dose toxicity study was performed by N. A. Littlefield et al.( Food Chem. Toxic. ,1989) to determine the oral toxic nature Gentian violet (548-62-9).

Combined repeated dose & carcinogenicity study was performed to determine the mutagenic nature of gentian violet. The study was performed using male and female Fischer F344 rats. Male and female weanling animals (F0) were randomly divided into four groups under barrier conditions and administered 0 (control), 100, 300 or 600 ppm (Males: 0, 30, 80 or 160 mg/Kg bw and females: 0, 40, 100 or 200 mg/Kg bw) GV in their feed for at least 80 days. In females, the body weights increased gradually throughout the study in 0, 30, 80 or 160 mg/Kg bw dosed group. However, the rate of increase was lower in the 160 mg/Kg bw group. After about 85 wk, the body weight of animals fed the 80 mg/Kg bw increased at a lower rate than the 30 mg/Kg bw and control groups. In males, at 200 mg/Kg bw dose group, a lower average body weight was noted than those for any other dose group. Body weights of male rats of 40 and 100 mg/Kg bw groups peaked at about 460g at about 70wk and then started a gradual decline at about 85 wk.

 253 rats were found to be in the moribund stage throughout the study period. At the end of the dosing period, the mortality rates in the females were 33, 38, 60 and 66% for the controls and 30, 80 and 160 mg/Kg dose groups, respectively. For males, the same respective dose groups had mortality rates after 104 wk of 33, 33, 48 and 39% for 0, 40, 100 or 200 mg/kg bw. The mortality in females was significantly different from the controls at the 0.001 level in the 80 mg/Kg bw (P = 0.00007) and 160 mg/Kg bw groups (P= 0.00005). In males, only the 100 mg/Kg bw (P = 0.0057) had a higher mortality than the control animals at the 0.05 levels.No non neoplastic effects related to administration of the test substance were observed at the 12- and 18-month necropsies in male and female rats. Most non-neoplastic lesions in the female rats that showed a dose response at 24 months were located in the liver. Lesions in the liver included eosinophilic foci, haematopoietic cell proliferation, mixed cell foci, regeneration, centrilobular necrosis and bile duct hyperplasia. In non-neoplastic lesions noted in the liver of male rats included clear cell foci, eosinophilic foci, mixed cell loci, regeneration and centrilobular necrosis. Lesions in other organs included follicle cyst of the thyroid gland, red pulp hyperplasia of the spleen and hyperplasia of the mesenteric lymph nodes. Based on the observations made, the No Observed Adverse Effect level (NOAEL) for Gentian violet in male and female rats is considered to be 40 and 30 mg/Kg bw respectively.

Repeated inhalation study:

According to column 2 of REACH Annex VIII, the acute toxicity inhalation study need not be conducted because exposure of humans via inhalation route is not likely taking into account the low vapour pressure of the substance N-phenyl-4-[[4-(phenylamino)phenyl][4-(phenylimino)cyclohexa-2,5-dien-1-ylidene]methyl]aniline monohydrochloride, which is reported as 8.36E-19 Pa. Also considering the particle size distribution of the substance the majority of the particles were found to be in the size of 150 micrometer which is much larger size range compared to the inhalable particulate matter. Thus, exposure to inhalable dust, mist and vapour of the chemical 1-hydroxybenzotriazole is highly unlikely. Therefore this study is considered for waiver. Waiver for repeated inhalation.

Repeated dermal study

The acute toxicity value for N-phenyl-4-[[4-(phenylamino)phenyl][4-(phenylimino)cyclohexa-2,5-dien-1-ylidene]methyl]aniline monohydrochloride (as provided in section 7.2.3) is >2000 mg/kg body weight. Also, given the use of the chemical as printing ink; repeated exposure by the dermal route is unlikely since the use of gloves is common practice in industries. Thus, it is expected that N-phenyl-4-[[4-(phenylamino)phenyl][4-(phenylimino)cyclohexa-2,5-dien-1-ylidene]methyl]aniline monohydrochloride (2152-64-9) shall not exhibit 28 day repeated dose toxicity by the dermal route. In addition, there is no data available that suggests that N-phenyl-4-[[4-(phenylamino)phenyl][4-(phenylimino)cyclohexa-2,5-dien-1-ylidene]methyl]aniline monohydrochloride (2152-64-9)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 read across and its prediction, N-phenyl-4-[[4-(phenylamino)phenyl][4-(phenylimino) cyclohexa-2,5-dien-1- ylidene]methyl]aniline monohydrochloride (2152-64-9) does not exhibit toxic nature upon repeated exposure by oral, inhalation and dermal route of exposure and hence is not likely to classify as toxicant.

Justification for classification or non-classification

Based on the data available for the target chemical and its prediction, N-phenyl-4-[[4-(phenylamino)phenyl][4-(phenylimino)cyclohexa-2,5-dien-1-ylidene]methyl]aniline monohydrochloride (2152-64-9) 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.