Registration Dossier

Administrative data

Description of key information

Repeated dose oral toxicity:

The Repeated Dose 28-day Oral Toxicity study was designed and conducted to determine the toxicity profile of the test chemical when administered daily for 28 days in the Sprague Dawley rats. The test chemical was dissolved in corn oil and was administered to animals at the dose levels of 0 mg/Kg, 25 mg/kg, 50 mg/kg and 100 mg/kg body weight. All the male and female animals from control and all the treateddose groups up to 100 mg/kg survived throughout the dosing period of 28 days. Reduced body weight gain of 8.55% and 11.56% was observed in male animals from 50 mg/kg and 100 mg/kg dose groups when compared with controls at the end of the dosing period of 28 days. Reduced body weight gain of 2.86% and 6.79% was observed in female animals from 50 mg/kg and 100 mg/kg dose groups when compared with controls at the end of the dosing period of 28 days. Although significant reduction in the body weight gain was observed in female animals from 50 mg/kg dose group, the effect was attributed to the biological variation within the test system and considered to be of no toxicological importance. Male and female animals from 25 mg/kg dose group exhibited normal body weight gain at the end of the dosing period of 28 days. Food intake of animals from control and different dose groups were found to be comparable throughout the dosing period of 28 days.Male and female animals treated at the dose levels of 100 mg/kg, exhibited violet discolouration of faces and anal region with semisolid faecal consistency during the dosing period of 28 days. Male and female animals from 25 mg/kg and 50 mg/kg dose groups exhibited no signs of toxicity throughout the dosing period of 28 days.Detailed clinical observations (weekly) did not reveal any signs of toxicity in all male and female animals from control and all the treated dose groups during the dosing period of 28 days.Towards the end of the exposure period in week 4, functional observation battery such as sensory reactivity to stimuli of different types (e.g. auditory, visual and proprioceptive stimuli) revealed no abnormalities attributable to the treatment. Grip strength values observed in male and female animals for control and different dose groups were comparable.Lower values of motor activity were observed in male animals from 25 mg/kg dose group for first interval and animals from 25 mg/kg and 50 mg/kg dose groups for second interval. Higher values of motor activity were observed in female animals from 25 mg/kg, 50 mg/kg and 100 mg/kg dose groups for first interval. These changes were within laboratory range and were considered to be of no toxicological importance. Haematological analysis performed on 29thday revealed statistically significant increase in the values of Total RBC of male rats dosed at 100 mg/kg and Total WBC of female rats dosed at 100 mg/kg. In addition statistically significant decrease was observed in the values of Total RBC and Platelets of male rats dosed at 25 mg/kg, Hb and HCT of male rats dosed at 25 mg/kg and 50 mg/kg, MCV and Total WBC of male rats dosed at 25 mg/kg, 50 mg/kg and 100 mg/kg and MCH of male rats dosed at 50 mg/kg and 100 mg/kg. The increase / decrease in the values of different parameters were marginal and within the normal laboratory limits. Although there was an increase/decrease in the values of various biochemical parameters as mentioned above, the deviations were marginal and within the range of normal laboratory limits. Organ weight data of male animals sacrificed on day 29, revealed increased relative weights of brain of animals from 50 mg/kg and 100 mg/kg dose groups, increased relative weights of liver of animals from 100 mg/kg dose group and increased relative weights of lungs of animals from 50 mg/kg dose group. Organ weight data of female animals sacrificed on day 29, revealed increased relative weights of liver of animals from 100 mg/kg dose group. Although significant change in organ weights were observed in animals from different dose groups, no related gross pathological or histological changes were seen and hence considered to be of no toxicological importance. Gross pathological examination was conducted in all animals when sacrificed terminally revealed test item coloured (blue) digestive tract that was evident from stomach to rectum region in four male animals from 50 mg/kg dose group and all male and female animals from 100 mg/kg dose group. In addition, the test item coloured (blue) caecum was observed in all male and five female animals from 25 mg/kg dose group and two male and five female animals from 50 mg/kg dose group. Histopathological examination conducted in rats of control and high dose groups did not reveal any abnormality attributable to the treatment. Based on the observations made, the No Observed Adverse Effect Level (NOAEL) of the test chemical in the Sprague Dawley rat via oral route, was found to be 25 mg/kg body weight in male and female animals.

Repeated dose Inhalation toxicity:

4,4'-bis(dimethylamino)-4''-(methylamino)trityl alcohol(CAS no 561-41-1)has very low vapor pressure (0.00000000052 Paat 25˚C), so the potential for the generation of inhalable vapours is very low.The particle size distribution was determined to be in the range of53 to 250 micrometer.Also the normal conditions of use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalatory route will be highly unlikely and therefore this end point was considered for waiver

Repeated dose dermal toxicity:

The acute dermal toxicity value for4,4'-bis(dimethylamino)-4''-(methylamino)trityl alcohol (CAS no 561-41-1) (as provided in section 7.2.3) is >2000 mg/kg body weight. Considering this, the end point for repeated dermal toxicity is considered as waiver.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
18-06- 2014 - 25-08-2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Data is from study report
Qualifier:
according to
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents)
Principles of method if other than guideline:
Repeated dose oral toxicity study was performed to determine the toxic nature of the test chemical
GLP compliance:
yes
Species:
rat
Strain:
Sprague-Dawley
Details on species / strain selection:
1) In order to meet the regulatory requirement for testing in a rodent species;

2) Widely used in industry as a species of choice for pre-clinical toxicological studies.

3) This strain is widely used throughout the industry in the non-clinical laboratory studies.

4) This study is intended to provide information on the health hazards likely to arise from short term accidental exposure to the test item by the oral route
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: National Institute of Biosciences, Pune.

- Age at study initiation: 6 to 8 weeks old.
- Weight at study initiation:
Male 168.08 g
Female 142.83 g

- Fasting period before study: No data available

- Housing: Animal was housed in polycarbonate cages. Three rats of same sex were housed together in each cage of size 39 cm X 28 cm X 14 cm. Paddy husk was used as bedding material.

- Diet (e.g. ad libitum): Rodent feed supplied by the Nutrivet Life Sciences, Pune, was provided ad libitum from individual feeders on cage top.

- Water (e.g. ad libitum): Water was provided ad libitum from individual bottles attached to the cages. Water was from a local source and passed through the reverse osmosis membrane before use.

- Acclimation period: 5 days prior to dosing.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3°C (actual range: 19.7 °C to 24.1 °C)

- Humidity (%):30% to 70% (actual range: 52.8% to 59.7%).

- Air changes (per hr): Ten air changes per hour of 100% fresh air that has been passed through the HEPA filters.

- Photoperiod (hrs dark / hrs light): An artificial light and dark cycle of 12 hours each were provided.


Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS: The test chemical was dissolved in corn oil for preparation of dosing solution(s). The solution(s) were prepared at concentrations of 0, 2.5, 5 and 10 mg/ml such that dosage of 0 (vehicle), 25, 50 and 100 mg/kg body weight

DIET PREPARATION
- Rate of preparation of diet (frequency): No data available

- Mixing appropriate amounts with (Type of food): No data available

- Storage temperature of food: No data available

VEHICLE
- Justification for use and choice of vehicle (if other than water): No data available

- Concentration in vehicle: 0, 25 mg/kg, 50 mg/kg and 100 mg/kg body weight

- Amount of vehicle (if gavage): 10 mL/Kg bw

- Lot/batch no. (if required): No data available

- Purity: No data available

Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Analysis for concentration and stability of the test chemical were conducted at Subcontracted Laboratory. Test item formulation samples prepared day 1 (pre-dosing) were sent to Subcontracted Laboratory.
Duration of treatment / exposure:
28 days consecutively
Frequency of treatment:
Once daily
Remarks:
Doses / Concentrations:
0 (vehicle), 25, 50 and 100 mg/kg body weight/day.
Basis:
actual ingested
No. of animals per sex per dose:
Control: 6 male, 6 female
250 mg/kg bw/day: 6 male, 6 female
500 mg/kg bw/day: 6 male, 6 female
100 0mg/kg bw/day: 6 male, 6 female
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale:
The doses were selected based on the results of the Dose Range Finder study conducted.
Salient features of the Dose Range Finding study were as follows :
The study was conducted at the dose levels of 0 mg/kg, 50 mg/kg, 100 mg/kg, 250 mg/kg, 500 mg/kg and 1000 mg/kg body weight.

1) Male and female animals from control, 50 mg/kg and 100 mg/kg dose groups survived through the dosing period of 14 days. All male and female animals treated at the dose levels of 250 mg/kg, 500 mg/kg and 1000 mg/kg died during the dosing period of 14 days.

2) Male and female animals from control, 50 mg/kg and 100 mg/kg dose groups exhibited normal body weight gain at the end of the dosing period of 14 days.

3) Daily clinical observations did not reveal any signs of toxicity in male and female animals from 50 mg/kg and 100 mg/kg dose groups during the dosing period of 14 days. Male and female animals treated at the dose levels of 250 mg/kg, 500 mg/kg and 1000 mg/kg, exhibited polyurea, diarrhoea, reduced locomotor activity, ataxic gait and distension during the dosing period.

4) Gross pathological examination did not reveal any abnormality attributable to the treatment in male and female animals from 50 mg/kg and 100 mg/kg dose groups.
Gross pathological examination revealed blue colouration to digestive tract from stomach to rectum and/or slight congestion in lungs in found dead animals in male and female animals from 250 mg/kg, 500 mg/kg and 1000 mg/kg dose groups.

Based on these results, the 28 day study dose levels were finalized as 0 mg/kg, 25 mg/kg, 50 mg/kg and 100 mg/kg body weight and animals were exposed to the treatment, every day, for a period of 28 days.

- Rationale for animal assignment (if not random): Animals were randomized by body weight.

- Rationale for selecting satellite groups: No data available

- Post-exposure recovery period in satellite groups: No data available

- Section schedule rationale (if not random): No data available
Positive control:
No data
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes

- Time schedule: Twice daily

- Cage side observations checked in table [No.?] were included. : Rats were observed for Behavior, Alterations, Vocalizations, Respiration and Palpebral closure. Mortality

DETAILED CLINICAL OBSERVATIONS: Yes

- Time schedule: Once before the start of dose administration and at least once a week thereafter until scheduled sacrifice. Handling observations Open field observations, Sensory Reactivity Observations were also performed

BODY WEIGHT: Yes

- Time schedule for examinations: On the day of randomization, first day of dosing, weekly thereafter and a fasting body weight at scheduled sacrifice on day 29.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes, The quantity of feed consumed by control and different treatment groups was recorded weekly until scheduled sacrifice

- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data available

- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data available

FOOD EFFICIENCY: No data available
- 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: No data available

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data available

- Time schedule for examinations: No data available

OPHTHALMOSCOPIC EXAMINATION: Yes

- Time schedule for examinations: Once a day

- Dose groups that were examined: 0 mg/kg, 250 mg/kg, 500 mg/kg and 1000 mg/kg bw/day.

HAEMATOLOGY: Yes, By using Beckman Coulter haematology analyzer.
- Time schedule for collection of blood: At termination.

- Anaesthetic used for blood collection: No data available

- Animals fasted: Yes, overnight fasted prior to sampling.

- How many animals: Blood collected from all rats of 0 mg/kg, 250 mg/kg, 500 mg/kg and 1000 mg/kg bw/day group at termination.

- Parameters checked in table [No.?] were examined. : Hemoglobin, Red Blood Corpuscles, Hematocrit, Mean Corpuscular Volume, Mean Corpuscular Hemoglobin, Mean Corpuscular Hemoglobin Concentration, Platelets, White Blood Corpuscles, Neutrophils, Lymphocytes, Eosinophils, Monocytes, Basophil and Prothrombin time were checked

CLINICAL CHEMISTRY: Yes, By using Dimension XpandPlus and Acculyte 5P.

- Time schedule for collection of blood: At termination.

- Animals fasted: Yes, overnight fasted prior to sampling.

- How many animals: Blood collected from all rats of 0 mg/kg, 250 mg/kg, 500 mg/kg and 1000 mg/kg bw/day group at termination.

- Parameters checked in table [No.?] were examined. : Total Protein, Blood Urea Nitrogen, Urea Nitrogen, Alanine Aminotransferase, Aspartate Aminotransferase, Alkaline Phosphatase, Gamma Glutamyl Transferase, Glucose, Calcium, Phosphorous, Albumin, Total Bilirubin, Creatinine , Total Cholesterol, Triglycerides, Globulin Calculated. Sodium, Potassium, Chloride were checked

URINALYSIS: No data available
- Time schedule for collection of urine: No data available

- Metabolism cages used for collection of urine: No data available

- Animals fasted: No data available

- Parameters checked in table [No.?] were examined. : No data available

NEUROBEHAVIOURAL EXAMINATION: - No data available

Time schedule for examinations: No data available

- Dose groups that were examined: No data available

- Battery of functions tested: sensory activity / grip strength / motor activity / other: Yes

OTHER:
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
All the animals of 0 mg/kg, 250 mg/kg, 500 mg/kg and 1000 mg/kg/bw/day group were sacrificed by CO2 asphyxiation and gross lesions were noted. Liver, Kidneys, Adrenals, Epididymides, Prostate + Seminal Vesicle with Coagulation gland as whole, Thymus, Spleen, Brain, Heart, Lungs, Uterus, Testes/Ovaries were dissected free of fat and weighed. The paired organs were weighed together.

From each rat, samples or the whole of the tissue were preserved. All tissues were fixed in 10% neutral buffered formalin except, eyes and testes of all animals were preserved in Davidson’s solution for 24 hours and transferred to 10% neutral buffered formalin. Procedure for preparation of slides of tissues of various organs from the rats of various dose groups was performed as per the standard operating procedures

HISTOPATHOLOGY: Yes
Control and treated at the highest dose level of 1000 mg/kg were subjected to sacrifice.
Organs examined: Adrenals, Aorta, Brain (cerebrum, cerebellum and pons), Caecum, Cervix, Colon, Duodenum, Epididymides, Eyes, Heart, Ileum, Jejunum, Kidneys, Liver, Lungs, Mesenteric Lymphnodes, Oesophagus, Ovaries, Pancreas, Pituitary gland, Pharyngeal Lymphnodes, Prostate, Rectum, Skeletal Muscles, Skin with Mammary Gland, Spleen, Sternum with bone marrow, Sciatic Nerve, Spinal Cord (Cervical, mid thoracic and lumbar), Stomach, Seminal Vesicles with Coagulation Gland, Testes, Thymus, Thyroid, Trachea, Vagina, Urinary Bladder, Uterus.
Statistics:
Adrenals, Aorta, Brain (cerebrum, cerebellum and pons), Caecum, Cervix, Colon, Duodenum, Epididymides, Eyes, Heart, Ileum, Jejunum, Kidneys, Liver, Lungs, Mesenteric Lymphnodes, Oesophagus, Ovaries, Pancreas, Pituitary gland, Pharyngeal Lymphnodes, Prostate, Rectum, Skeletal Muscles, Skin with Mammary Gland, Spleen, Sternum with bone marrow, Sciatic Nerve, Spinal Cord (Cervical, mid thoracic and lumbar), Stomach, Seminal Vesicles with Coagulation Gland, Testes, Thymus, Thyroid, Trachea, Vagina, Urinary Bladder, Uterus.
Clinical signs:
no effects observed
Description (incidence and severity):
Male -
Group I (0 mg/kg): No clinical signs of toxicity were observed in the animals throughout the dosing period of 28 days (animal nos.1 to 6).
Group II (25 mg/kg): No clinical signs of toxicity were observed in the animals throughout the dosing period of 28 days (animal nos.13 to 18).
Group III (50 mg/kg): No clinical signs of toxicity were observed in the animals throughout the dosing period of 28 days (animal nos.25 to 30).
Group IV (100 mg/kg): Violet discolouration of faces and anal region with semisolid faecal consistency were observed in all animals (animal nos.37 to 42, with onset from day 13) during the dosing period of 28 days.
Female -
Group I (0 mg/kg): No clinical signs of toxicity were observed in the animals throughout the dosing period of 28 days (animal nos.7 to 12).
Group II (25 mg/kg): No clinical signs of toxicity were observed in the animals throughout the dosing period of 28 days (animal nos.19 to 24).
Group III (50 mg/kg): No clinical signs of toxicity were observed in the animals throughout the dosing period of 28 days (animal nos.31 to 36).
Group IV (100 mg/kg): Violet discolouration of faces and anal region with semisolid faecal consistency were observed in all animals (animal nos.43 to 48, with onset from day 12) during the dosing period of 28 days.
Mortality:
no mortality observed
Description (incidence):
All the animals from control and all the treated dose groups survived throughout the dosing period of 28 days
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Male -
Animals from control and low dose group exhibited normal body weight gain throughout the dosing period of 28 days.
Reduced body weight gain of 8.55% and 11.56% was observed in animals from 50 mg/kg and 100 mg/kg dose groups when compared with controls at the end of the dosing period of 28 days (p≤0.01).
Female -
Animals from control and low dose group exhibited normal body weight gain throughout the dosing period of 28 days.
Reduced body weight gain of 2.86% and 6.79% was observed in animals from 50 mg/kg and 100 mg/kg dose groups when compared with controls at the end of the dosing period of 28 days (p≤0.01).

Although significant reduction in the body weight gain was observed, the effect was attributed to the biological variation within the test system and considered to be of no toxicological importance.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
Animals from control and all treated dose groups exhibited normal feed consumption at the end of the dosing period of 28 days.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
no effects observed
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
Male and Female -
Haematological investigations conducted at the end of dosing period on day 29, revealed following significant changes in the values of different parameters studied when compared with that of respective controls.
Male :
Total RBC : Increased values were obtained for animals from 100 mg/kg dose group (p≤0.05),
Total RBC and Platelets : Decreased values were obtained for animals from 25 mg/kg dose group (p≤0.01),
Hb : Decreased values were obtained for animals from 25 mg/kg (p≤0.01) and 50 mg/kg (p≤0.05) dose groups,
HCT : Decreased values were obtained for animals from 25 mg/kg and 50 mg/kg dose groups (p≤0.01),
MCV and Total WBC : Decreased values were obtained for animals from 25 mg/kg, 50 mg/kg and 100 mg/kg dose groups (p≤0.01) and
MCH : Decreased values were obtained for animals from 50 mg/kg and 100 mg/kg dose groups (p≤0.05).
Female :
Total WBC : Increased values were obtained for animals from 100 mg/kg dose group (p≤0.01).
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
Male and Female -
Biochemical investigations conducted at the end of dosing period on day 29, revealed following significant changes in the values of different parameters studied when compared with that of respective controls.
Male :
Alanine Aminotransferase : Elevated levels were observed in animals from 100 mg/kg dose group (p≤0.05),
Sodium, Potassium and Chloride : Elevated levels were observed in animals from 25 mg/kg, 50 mg/kg and 100 mg/kg dose groups (p≤0.01) and
Calcium : Decreased levels were observed in animals from 25 mg/kg, 50 mg/kg and 100 mg/kg dose groups (p≤0.01).

Female :
Calcium : Elevated levels were observed in animals from 25 mg/kg dose group (p≤0.05) and
Chloride : Decreased levels were observed in animals from 25 mg/kg (p≤0.01), 50 mg/kg (p≤0.01) and 100 mg/kg (p≤0.05) dose groups.
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
Male -
In comparison with controls organ weight data of male animals sacrificed on day 29, revealed increased relative weight of brain of animals from 50 mg/kg and 100 mg/kg dose groups (p≤0.01), increased relative weight of liver of animals from 100 mg/kg dose group (p≤0.05) and increased relative weights of lungs of animals from 50 mg/kg dose group.

Female -
In comparison with controls organ weight data of female animals sacrificed on day 29, revealed increased relative weight of liver of animals from 100 mg/kg dose group (p≤0.01).

Although significant change in organ weights were observed in animals from different dose groups, no related gross pathological or histological changes were seen and hence considered to be of no toxicological importance.
Gross pathological findings:
no effects observed
Description (incidence and severity):
Gross pathological examination was conducted in all animals when sacrificed terminally.
Test item coloured (blue) digestive tract was evident from stomach to rectum region in four male animals from 50 mg/kg dose group and all male and female animals from 100 mg/kg dose group. In addition, the test item coloured (blue) caecum was observed in all male and five female animals from 25 mg/kg dose group and two male and five female animals from 50 mg/kg dose group.
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
No treatment related histopathological changes were evident in male and female rats from control and high dose groups.
Histopathological examination revealed minimal focal to multifocal periportal mononuclear cell infiltration in the liver; minimal interstitial haemorrhages and/or mononuclear cells infiltration and/or tubular dilatation in the kidneys; minimal alveolar haemorrhages and/or alveolar histiocytosis in the lungs; minimal multifocal haemosiderosis and/or diffused congestion in spleen; minimal luminal dilatation and/or endometrial gland dilatation in uterus; minimal luminal seminal coagulum in the urinary bladder; minimal dilatation of zona reticularis and/or vacuolation in zona fasiculata in adrenals; minimal multifocal haemorrhages in thymus; presence of ultimobranchial cysts in thyroid; in male or female animals from control and high dose group. All the changes observed in the control and high dose treatment group animals were similar, incidental and mode of death related, physiological and are covered in the facility historical data of the histopathology findings.
Histopathological findings: neoplastic:
no effects observed
Description (incidence and severity):
No treatment related histopathological changes were evident in male and female rats from control and high dose groups.
Histopathological examination revealed minimal focal to multifocal periportal mononuclear cell infiltration in the liver; minimal interstitial haemorrhages and/or mononuclear cells infiltration and/or tubular dilatation in the kidneys; minimal alveolar haemorrhages and/or alveolar histiocytosis in the lungs; minimal multifocal haemosiderosis and/or diffused congestion in spleen; minimal luminal dilatation and/or endometrial gland dilatation in uterus; minimal luminal seminal coagulum in the urinary bladder; minimal dilatation of zona reticularis and/or vacuolation in zona fasiculata in adrenals; minimal multifocal haemorrhages in thymus; presence of ultimobranchial cysts in thyroid; in male or female animals from control and high dose group. All the changes observed in the control and high dose treatment group animals were similar, incidental and mode of death related, physiological and are covered in the facility historical data of the histopathology findings.
Other effects:
not specified
Details on results:
Clinical signs and mortality
Clinical signs :
Daily clinical observations did not reveal any signs of toxicity in male and female animals from of 0 mg/kg, 250 mg/kg, 500 mg/kg and 1000 mg/kg bw/day dose groups during the dosing period of 28 days.

Mortality:
No mortality were observed in any of the traeted groups of 0 mg/kg, 250 mg/kg, 500 mg/kg and 1000 mg/kg bw/day.

Body weight and weight gain
All the of 0 mg/kg, 250 mg/kg, 500 mg/kg and 1000 mg/kg bw/day dose groups exhibited normal body weight gain at the end of the study period of 28 days.

Food consumption and compound intake
Food consumption :
Food intake of animals from control and 250 mg/kg, 500 mg/kg and 1000 mg/kg bw/day dose groups were found to be normale throughout the study period of 28 days.

Food efficiency
No data available

Water consumption and compound intake
No data available

Opthalmoscopic examination
No data available

Haematology
Statistically significant increase in the values of MCHC of male rats dosed at 250 mg/kg/bw/day and 1000 mg/kg/bw/day. In addition statistically significant decrease was observed in the values of Hb and HCT of male rats dosed at 1000 mg/kg. The increase / decrease in the values of different parameters were marginal and within the normal laboratory limits.
No significant changes were observed in the values of different parameters studied in female animals from different dose groups when compared with that of controls.

Clinical chemistry

Increase level of Calcium and Sodium in male rats dosed at 250 mg/kg, 500 mg/kg and 1000 mg/kg of 4,4’-Methylenebis (2,6-dimethylphenol).

Significant increase level of Cholesterol levels in male rats dosed at 250 mg/kg of 4,4’-Methylenebis (2,6-dimethylphenol).

Decrease level of Phosphorous in male rats dosed at 1000 mg/kg of 4,4’-Methylenebis (2,6-dimethylphenol) were observed.

Decrease level of Chloride in male rats dosed at 250 mg/kg, 500 mg/kg and 1000 mg/kg.

Decrease level of Glucose in female rats dosed at 500 mg/kg of 4,4’-Methylenebis (2,6-dimethylphenol) was observed.

Although there was an increase/decrease in the values of various biochemical parameters are within the normal range of limits.

Urinanalysis
No data available

Neurobehaviour
No data available

Organ weights
Female animals revealed increased relative weights of liver of animals from 250 mg/kg and 500 mg/kg dose groups, increased relative weights of kidneys of animals from 250 mg/kg dose group and decreased relative weights of spleen of animals from 1000 mg/kg dose group.

Although significant change in organ weights were observed in animals from different dose groups, but the effect was not due to 4,4’-Methylenebis (2,6-dimethylphenol).

Gross pathology
In male and female animals from control and 250 mg/kg/bw/day, 500 mg/kg/bw/day and 1000 mg/kg/bw/day of test item. All the treatment groups did not reveal any abnormality.

Histopathology
Focal to multifocal periportal mononuclear cell infiltration in the liver; minimal interstitial haemorrhages and/or tubular dilatation in the kidneys; minimal alveolar haemorrhages and/or alveolar histiocytosis in the lungs; minimal multifocal haemosiderosis and/or diffused congestion in spleen; minimal eosinophilic infiltration and/or luminal dilatation and/or endometrial gland dilatation in uterus; minimal dilatation of zona reticularis and/or presence of accessory adrenocortical tissue in adrenals; minimal multifocal haemorrhages in thymus; presence of ultimobranchial cysts in thyroid; in male or female animals from control and high dose group. All the changes observed in the control and 1000 mg/kg/bw/day dose group animals were similar and no toxic effect of 4,4’-Methylenebis (2,6-dimethylphenol) were observed
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Based on Clinical biochemistry, Organ weight, Gross pathological and Histopathology.
Critical effects observed:
not specified
Conclusions:
The No Observed Adverse Effect Level (NOAEL) of the test chemical in the Sprague Dawley rat via oral route, was found to be 25 mg/kg body weight in male and female animals.
Executive summary:

The Repeated Dose 28-day Oral Toxicity study was designed and conducted to determine the toxicity profile of the test chemical when administered daily for 28 days in the Sprague Dawley rats. The test chemical was dissolved in corn oil and was administered to animals at the dose levels of 0 mg/Kg, 25 mg/kg, 50 mg/kg and 100 mg/kg body weight. All the male and female animals from control and all the treateddose groups up to 100 mg/kg survived throughout the dosing period of 28 days. Reduced body weight gain of 8.55% and 11.56% was observed in male animals from 50 mg/kg and 100 mg/kg dose groups when compared with controls at the end of the dosing period of 28 days. Reduced body weight gain of 2.86% and 6.79% was observed in female animals from 50 mg/kg and 100 mg/kg dose groups when compared with controls at the end of the dosing period of 28 days. Although significant reduction in the body weight gain was observed in female animals from 50 mg/kg dose group, the effect was attributed to the biological variation within the test system and considered to be of no toxicological importance. Male and female animals from 25 mg/kg dose group exhibited normal body weight gain at the end of the dosing period of 28 days. Food intake of animals from control and different dose groups were found to be comparable throughout the dosing period of 28 days.Male and female animals treated at the dose levels of 100 mg/kg, exhibited violet discolouration of faces and anal region with semisolid faecal consistency during the dosing period of 28 days. Male and female animals from 25 mg/kg and 50 mg/kg dose groups exhibited no signs of toxicity throughout the dosing period of 28 days.Detailed clinical observations (weekly) did not reveal any signs of toxicity in all male and female animals from control and all the treated dose groups during the dosing period of 28 days.Towards the end of the exposure period in week 4, functional observation battery such as sensory reactivity to stimuli of different types (e.g. auditory, visual and proprioceptive stimuli) revealed no abnormalities attributable to the treatment. Grip strength values observed in male and female animals for control and different dose groups were comparable.Lower values of motor activity were observed in male animals from 25 mg/kg dose group for first interval and animals from 25 mg/kg and 50 mg/kg dose groups for second interval. Higher values of motor activity were observed in female animals from 25 mg/kg, 50 mg/kg and 100 mg/kg dose groups for first interval. These changes were within laboratory range and were considered to be of no toxicological importance. Haematological analysis performed on 29thday revealed statistically significant increase in the values of Total RBC of male rats dosed at 100 mg/kg and Total WBC of female rats dosed at 100 mg/kg. In addition statistically significant decrease was observed in the values of Total RBC and Platelets of male rats dosed at 25 mg/kg, Hb and HCT of male rats dosed at 25 mg/kg and 50 mg/kg, MCV and Total WBC of male rats dosed at 25 mg/kg, 50 mg/kg and 100 mg/kg and MCH of male rats dosed at 50 mg/kg and 100 mg/kg. The increase / decrease in the values of different parameters were marginal and within the normal laboratory limits. Although there was an increase/decrease in the values of various biochemical parameters as mentioned above, the deviations were marginal and within the range of normal laboratory limits. Organ weight data of male animals sacrificed on day 29, revealed increased relative weights of brain of animals from 50 mg/kg and 100 mg/kg dose groups, increased relative weights of liver of animals from 100 mg/kg dose group and increased relative weights of lungs of animals from 50 mg/kg dose group. Organ weight data of female animals sacrificed on day 29, revealed increased relative weights of liver of animals from 100 mg/kg dose group. Although significant change in organ weights were observed in animals from different dose groups, no related gross pathological or histological changes were seen and hence considered to be of no toxicological importance. Gross pathological examination was conducted in all animals when sacrificed terminally revealed test item coloured (blue) digestive tract that was evident from stomach to rectum region in four male animals from 50 mg/kg dose group and all male and female animals from 100 mg/kg dose group. In addition, the test item coloured (blue) caecum was observed in all male and five female animals from 25 mg/kg dose group and two male and five female animals from 50 mg/kg dose group. Histopathological examination conducted in rats of control and high dose groups did not reveal any abnormality attributable to the treatment. Based on the observations made, the No Observed Adverse Effect Level (NOAEL) of the test chemical in the Sprague Dawley rat via oral route, was found to be 25 mg/kg body weight in male and female animals.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
25 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
Data is from study report

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records
Reference
Endpoint:
repeated dose toxicity: inhalation, other
Data waiving:
other justification
Justification for data waiving:
other:
Endpoint conclusion
Endpoint conclusion:
no study available
Quality of whole database:
Waiver

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Link to relevant study records
Reference
Endpoint:
repeated dose toxicity: dermal, other
Data waiving:
other justification
Justification for data waiving:
other:
Endpoint conclusion
Endpoint conclusion:
no study available
Quality of whole database:
Waiver

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Data available for the target chemical was reviewed to determine the toxic nature of the test chemical. The studies are as mentioned below:

Repeated dose oral toxicity:

The Repeated Dose 28-day Oral Toxicity study was designed and conducted to determine the toxicity profile of the test chemical when administered daily for 28 days in the Sprague Dawley rats. The test chemical was dissolved in corn oil and was administered to animals at the dose levels of 0 mg/Kg, 25 mg/kg, 50 mg/kg and 100 mg/kg body weight. All the male and female animals from control and all the treateddose groups up to 100 mg/kg survived throughout the dosing period of 28 days. Reduced body weight gain of 8.55% and 11.56% was observed in male animals from 50 mg/kg and 100 mg/kg dose groups when compared with controls at the end of the dosing period of 28 days. Reduced body weight gain of 2.86% and 6.79% was observed in female animals from 50 mg/kg and 100 mg/kg dose groups when compared with controls at the end of the dosing period of 28 days. Although significant reduction in the body weight gain was observed in female animals from 50 mg/kg dose group, the effect was attributed to the biological variation within the test system and considered to be of no toxicological importance. Male and female animals from 25 mg/kg dose group exhibited normal body weight gain at the end of the dosing period of 28 days. Food intake of animals from control and different dose groups were found to be comparable throughout the dosing period of 28 days.Male and female animals treated at the dose levels of 100 mg/kg, exhibited violet discolouration of faces and anal region with semisolid faecal consistency during the dosing period of 28 days. Male and female animals from 25 mg/kg and 50 mg/kg dose groups exhibited no signs of toxicity throughout the dosing period of 28 days.Detailed clinical observations (weekly) did not reveal any signs of toxicity in all male and female animals from control and all the treated dose groups during the dosing period of 28 days.Towards the end of the exposure period in week 4, functional observation battery such as sensory reactivity to stimuli of different types (e.g. auditory, visual and proprioceptive stimuli) revealed no abnormalities attributable to the treatment. Grip strength values observed in male and female animals for control and different dose groups were comparable.Lower values of motor activity were observed in male animals from 25 mg/kg dose group for first interval and animals from 25 mg/kg and 50 mg/kg dose groups for second interval. Higher values of motor activity were observed in female animals from 25 mg/kg, 50 mg/kg and 100 mg/kg dose groups for first interval. These changes were within laboratory range and were considered to be of no toxicological importance. Haematological analysis performed on 29thday revealed statistically significant increase in the values of Total RBC of male rats dosed at 100 mg/kg and Total WBC of female rats dosed at 100 mg/kg. In addition statistically significant decrease was observed in the values of Total RBC and Platelets of male rats dosed at 25 mg/kg, Hb and HCT of male rats dosed at 25 mg/kg and 50 mg/kg, MCV and Total WBC of male rats dosed at 25 mg/kg, 50 mg/kg and 100 mg/kg and MCH of male rats dosed at 50 mg/kg and 100 mg/kg. The increase / decrease in the values of different parameters were marginal and within the normal laboratory limits. Although there was an increase/decrease in the values of various biochemical parameters as mentioned above, the deviations were marginal and within the range of normal laboratory limits. Organ weight data of male animals sacrificed on day 29, revealed increased relative weights of brain of animals from 50 mg/kg and 100 mg/kg dose groups, increased relative weights of liver of animals from 100 mg/kg dose group and increased relative weights of lungs of animals from 50 mg/kg dose group. Organ weight data of female animals sacrificed on day 29, revealed increased relative weights of liver of animals from 100 mg/kg dose group. Although significant change in organ weights were observed in animals from different dose groups, no related gross pathological or histological changes were seen and hence considered to be of no toxicological importance. Gross pathological examination was conducted in all animals when sacrificed terminally revealed test item coloured (blue) digestive tract that was evident from stomach to rectum region in four male animals from 50 mg/kg dose group and all male and female animals from 100 mg/kg dose group. In addition, the test item coloured (blue) caecum was observed in all male and five female animals from 25 mg/kg dose group and two male and five female animals from 50 mg/kg dose group. Histopathological examination conducted in rats of control and high dose groups did not reveal any abnormality attributable to the treatment. Based on the observations made, the No Observed Adverse Effect Level (NOAEL) of the test chemical in the Sprague Dawley rat via oral route, was found to be 25 mg/kg body weight in male and female animals.

Repeated dose Inhalation toxicity:

4,4'-bis(dimethylamino)-4''-(methylamino)trityl alcohol(CAS no 561-41-1)has very low vapor pressure (0.00000000052 Paat 25˚C), so the potential for the generation of inhalable vapours is very low.The particle size distribution was determined to be in the range of53 to 250 micrometer.Also the normal conditions of use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalatory route will be highly unlikely and therefore this end point was considered for waiver

Repeated dose dermal toxicity:

The acute dermal toxicity value for4,4'-bis(dimethylamino)-4''-(methylamino)trityl alcohol (CAS no 561-41-1) (as provided in section 7.2.3) is >2000 mg/kg body weight. Considering this, the end point for repeated dermal toxicity is considered as waiver.

Based on the data available for the target chemical, the test chemical is not likely to be toxic upon repeated exposure by oral, dermal and inhalation route of exposure.

Justification for classification or non-classification

Based on the data available for the target chemical, the test chemical is not likely to be toxic upon repeated exposure by oral, dermal and inhalation route of exposure.