Registration Dossier

Administrative data

Description of key information

Repeated Dose toxicity: Oral

Based on the available results and applying the weight of evidence approach, the test chemical can be considered to be non-toxic to living organisms when dosed repeatedly via oral route. The No Observed Adverse Effect Level can be considered to be >1000 mg/kgbw/day. Comparing the above annotations with the criteria of CLP Regulation, the test chemical can be classified under the category “Not Classified”.

Repeated Dose toxicity: Inhalation

A short-term toxicity study need not be conducted as because exposure of humans via inhalation route in production and/or use is not likely based on the provided thorough and rigorous exposure.The estimated vapour pressure of the test chemical was 5.47E-14 mm Hg. Hence, this endpoint was considered for waiver.

Repeated Dose Toxicity: Dermal

A short-term toxicity study need not be conducted as because exposure of humans via dermal route in production and/or use is not likely based on the provided thorough and rigorous exposure assessment. The acute dermal toxicity value for the test chemical (as provided in section 7.2.3) is >2000 mg/kg body weight. The substance failed to cause any dermal irritation as well sensitization. Considering this, the end point for repeated dermal toxicity is considered as waiver.

Key value for chemical safety assessment

Toxic effect type:
dose-dependent

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: oral
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Remarks:
Weight of evidence approach based on various test chemicals
Justification for type of information:
Weight of evidence approach based on various test chemicals
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
other: Weight of evidence approach based on various test chemicals
Principles of method if other than guideline:
Weight of evidence approach based on various test chemicals
GLP compliance:
not specified
Limit test:
no
Species:
rat
Strain:
other: 2.albino ; 3.Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
Not specified
Route of administration:
oral: gavage
Vehicle:
other: 2. ; 3.corn oil
Details on oral exposure:
2. PREPARATION OF DOSING SOLUTIONS: The test chemical was mixed with diet at dose levels of 0, 0.25, 1.0 or 3.0 % (0, 100, 400, 1200 mg/kg/day)
DIET PREPARATION
- Rate of preparation of diet (frequency): No data
- Mixing appropriate amounts with (Type of food): No data
- Storage temperature of food: No data
VEHICLE
- Justification for use and choice of vehicle (if other than water): Diet
- Concentration in vehicle: 0, 0.25, 1.0 or 3.0 % (0, 100, 400, 1200 mg/kg/day)
- Amount of vehicle (if gavage): No data
- Lot/batch no. (if required): No data
- Purity: No data
3. The test item was diluted with corn oil for preparation of solution(s).The solution(s) of the test chemica were made at volumes suitable for daily use for 28 days. The solution(s) were prepared at concentrations of 0, 25, 50 and 100 mg/ml such that dosage of 0 (vehicle), 250, 500 and 1000 mg/kg body weight respectively were administered.
VEHICLE
- Justification for use and choice of vehicle (if other than water): Corn oil
- Concentration in vehicle: 0 (vehicle), 250, 500 and 1000 mg/kg body weight
- Amount of vehicle (if gavage): upto 10 ml/kg body weight
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
2.
3. Stability determined by UV
Duration of treatment / exposure:
2. 2 years
3. 28 days
Frequency of treatment:
2. Daily
3. daily
Remarks:
Study 2: 0, 0.25, 1.0 or 3.0 % (0, 100, 400, 1200 mg/kg/day)
Remarks:
Study 3. 0, 250, 500, 1000 mg/kg/day (actual dose received)
No. of animals per sex per dose:

2. Total: 160 males and 160 females
0 mg/Kg/day: 80 males and 80 females
100 mg/Kg/day: 25 males and 25 females
400 mg/Kg/day: 25 males and 25 females
1200 mg/Kg/day: 25 males and 25 females
3. Total: 72
0 mg/kg bw: 6 male, 6 female
250 mg/kg bw: 6 male, 6 female
500 mg/kg bw: 6 male, 6 female
1000 mg/kg bw: 6 male, 6 female
Reversal group
0 mg/kg bw: 6 male, 6 female
1000 mg/kg bw: 6 male, 6 female
Control animals:
yes, concurrent vehicle
Details on study design:
3. - Dose selection rationale:
- Rationale for animal assignment (if not random):The animals of uniform body weight were selected.The individual body weights of the animals did not exceed ± 20% of group mean body weight. The group means body weights of all the groups were approximately equal.
Positive control:
Not specified
Observations and examinations performed and frequency:
2. CAGE SIDE OBSERVATIONS: Yes
- Time schedule: No data
- Cage side observations checked in table [No.?] were included. Appearance and behavior of test rats, mortality
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): Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data
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: No data
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data
- Time schedule for examinations: No data
OPHTHALMOSCOPIC EXAMINATION: No data
- Time schedule for examinations: No data
- Dose groups that were examined: No data
HAEMATOLOGY: Yes
- Time schedule for collection of blood: No data
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: No data
- Parameters checked in table [No.?] were examined. Hematocrit and hemoglobin values
CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: No data
- Animals fasted: No data
- How many animals: No data
- Parameters checked in table [No.?] were examined. Bilirubunuria
URINALYSIS: Yes
- Time schedule for collection of urine: No data
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data
- Parameters checked in table [No.?] were examined. No data
NEUROBEHAVIOURAL EXAMINATION: No data
- Time schedule for examinations: No data
- Dose groups that were examined: No data
- Battery of functions tested: sensory activity / grip strength / motor activity / other: No data
OTHER: No data

3. CAGE SIDE OBSERVATIONS: Yes
- Time schedule:twice daily
- Cage side observations checked in table [No.?] were included. Viability were observed
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule:daily
BODY WEIGHT: Yes
- Time schedule for examinations:Body weights were recorded on the day of randomization, day of first dosing, weekly thereafter and a fasting body weight at scheduled sacrifice on day 29 and day 43.
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes
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: Yes
- Time schedule for examinations: prior to the initiation of the dosing and at scheduled sacrifice.
- Dose groups that were examined: All dose group were examined.
HAEMATOLOGY: Yes
- Time schedule for collection of blood:At the end of dosing period on day 29 and at termination of recovery period on day 43.
- Anaesthetic used for blood collection: Not specified
- Animals fasted: Yes, overnight
- How many animals: All 72 animals were examined.
- Parameters checked in table [No.?] were examined.Hemoglobin (g/dL), Red Blood Corpuscles (x106 /μL), Hematocrit (%), Mean Corpuscular Volume (fL), Mean Corpuscular Hemoglobin (pg), Mean
Corpuscular Hemoglobin Concentration (g/dL), Platelets (x 103 /μL), White Blood Corpuscles (x103 /μL), Reticulocytes (%), Neutrophils (%), Lymphocytes (%), Eosinophils (%), Monocytes (%), Basophil (%) and Prothrombin time (sec.) were examined.
CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood:At the end of dosing period on day 29 and at termination of recovery period on day 43.
- Anaesthetic used for blood collection: Not specified
- Animals fasted: Yes, overnight
- How many animals: All 72 animals were examined.
- Parameters checked in table [No.?] were examined.: Total Protein (g/dL), Blood Urea Nitrogen (mg/dL), Urea Nitrogen (mg/dL) Calculated, Alanine Aminotransferase (U/L), Aspartate Aminotransferase(U/L), Alkaline Phosphatase (U/L), Gamma Glutamyl Transferase (U/L), Glucose (mg/dL), Calcium(mmol/L), Phosphorous (mg/dL), Albumin (g/dL), Total Bilirubin (mg/dL), Creatinine (mmol/L), Total Cholesterol (mg/dL), Triglycerides (mg/dL), Globulin (g/dL) Calculated, Sodium (mmol/L), Potassium (mmol/L), Chloride (mmol/L) and Bile acid (mmol/L) were examined.
URINALYSIS: Yes
- Time schedule for collection of urine: during the last week of dosing period and on reversal group rats at termination of recovery period on day 43.
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Not specified
- Parameters checked in table [No.?] were examined: Volume, Appearance, Colour, pH, Specific Gravity, Proteins, Glucose, Ketones, Bilirubin, Urobililogen, Occult Blood and Nitrite were examined.
NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: Towards the end of the exposure period of 28 days and towards the end of the recovery period on day 42
- Dose groups that were examined:All dose group were examined.
- Battery of functions tested: sensory activity, grip strength, motor activity, Visual Placing Response were examined.
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
OTHER:Organ Weights were examined.
Sacrifice and pathology:
2. GROSS PATHOLOGY: Yes, any gross changes in the organs or viscera attributable to the test material were noted
HISTOPATHOLOGY: Yes
3. GROSS PATHOLOGY: Yes, after 28 consecutive days of oral administration, all surviving study rats were sacrificed on day 29 (Group I, III, IV, V). In addition all rats from reversal groups were sacrificed on day 43 (Group II and VI).
HISTOPATHOLOGY: Yes, From each rat, samples or the whole of the tissues listed below 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.

GROSS PATHOLOGY: Yes, the male animals were necropsied after 28 days of administration of test chemical.

HISTOPATHOLOGY: Yes , Animals were observed microscopically.
Statistics:
3. Raw data was processed and analyzed for reporting group means and standard deviations with significance between the controls and treated groups, using SYSTAT 13 validated statistical software supplied by Starcom Information Technology Limited, Bangalore developed by Systat Software, Inc. USA. All the parameters characterized by continuous data such as body weight, feed consumption (calculated as gram per animal), organ weight, relative organ weight, haematological and clinical chemistry data were subjected to Bartlett’s test to meet the homogeneity of variance before conducting Analysis of Variance (ANOVA) and Dunnett’s t-test. Where the data was not meet the homogeneity of variance, Student’s t-test were performed to calculate significance.Significance was calculated at 5% level and indicated in the summary tables as follows:
* = Significant than control at 95% level of confidence (p<0.05).
Clinical signs:
no effects observed
Description (incidence and severity):
2. Appearance and behavior of the test rats were generally comparable to those of the controls
3. Male -
Group I (Control, 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 (Control, 0 mg/kg, Reversal): No clinical signs of toxicity were observed in the animals throughout the dosing period of 28 days and during the post-dosing recovery period (animal nos.13 to
18).
Group III (250 mg/kg): Test item coloured faeces were observed in all animals (animal nos.25 to 30, with onset from day 2) during the dosing period of 28 days.
Group IV (500 mg/kg): Test item coloured faeces were observed in all animals (animal nos.37 to 42, with onset from day 2) during the dosing period of 28 days.
Group V (1000 mg/kg): Test item coloured faeces were observed in all animals (animal nos.49 to 54, with onset from day 2) during the dosing period of 28 days.
Group VI (1000 mg/kg, Reversal): Test item coloured faeces were observed in all animals (animal nos.61 to 66, with onset from day 2) throughout the dosing period of 28 days and during the post dosing recovery period.
Female -
Group I (Control, 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 (Control, 0 mg/kg, Reversal): No clinical signs of toxicity were observed in the animals throughout the dosing period of 28 days and during the post-dosing recovery period (animal nos.19 to 24).
Group III (250 mg/kg): Test item coloured faeces were observed in all animals (animal nos.31 to 36, with onset from day 2) during the dosing period of 28 days
Group IV (500 mg/kg): Test item coloured faeces were observed in all animals (animal nos.43 to 48, with onset from day 2) during the dosing period of 28 days
Group V (1000 mg/kg): Test item coloured faeces were observed in all animals (animal nos.55 to 60, with onset from day 2) during the dosing period of 28 days. No significant effect was observed at dose level of 0,100, 300 and 1000 mg/kg bw/day in treated group compare to control.
Group VI (1000 mg/kg, Reversal): Test item coloured faeces were observed in all animals (animal nos.67 to 70, with onset from day 2) throughout the dosing period of 28 days and during the post-dosing recovery period
Before commencement of treatment:
In home cage observation, rat from different dose groups and control group revealed normal behavior, alterations, vocalization, respiration and palpebral closer.During handling observation, handling of rats did not reveal any abnormality from different dose groups and control group. In the open field observation, rat did not reveal any abnormality from different dose groups and control group.
During treatment:
In home cage observation, rat from different dose groups and control group revealed normal behavior, alterations, vocalization, respiration and palpebral closer.During handling observation, handling of rats did not reveal any abnormality from different dose groups and control group.In the open field observation, rat did not reveal any abnormality from different dose groups and control group.Detailed clinical observation did not reveal any abnormality in all groups during the dosing period of 28 days and during the post-dosing recovery period.
Mortality:
no mortality observed
Description (incidence):
2. No mortality was observed
3. All animals from control and different dose groups survived throughout the dosing period of 28 days and the post-dosing recovery period of 14 days.
No significant effect was observed at dose level of 0,100, 300 and 1000 mg/kg bw/day in treated group compare to control.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
3. Male -
Animals from control and different dose groups exhibited normal body weight gain throughout the dosing period of 28 days. Reduced body weight gain of 5.48% was observed in male animals from 1000 mg/kg dose group and this effect was attributed to the biological variation within the test system and considered to be of no toxicological importance. During the post-dosing recovery period, animals from 1000 mg/kg reversal group exhibited normal body weight gain when compared with that of control animals.
Female -
Animals from control and different dose groups exhibited normal body weight gain throughout the dosing period of 28 days.During the post-dosing recovery period, animals from 1000 mg/kg reversal group exhibited normal body weight gain when compared with that of control animals.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
2. At the 3% level (1200 mg/Kg/day), food consumption was significant lower than controls for the first six months but comparable to controls during the remainder of the study
3. Male and Female -
Animals from control and different dose groups exhibited normal feed consumption at the end of the dosing period of 28 days.Animals from control reversal and high reversal dose groups exhibited normal feed consumption at the end of the recovery period of 14 days.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
no effects observed
Description (incidence and severity):
3. No ocular abnormalities were observed on ophthalmological examination in the animals during preexposure and at the end of the respective termination.
Haematological findings:
no effects observed
Description (incidence and severity):
2. Significantly lower hematocrit and hemoglobin values in comparison with controls were obtained at all time intervals except 24 months for the males fed 3%. No such effect was noted in the females
3. Male and Female -
Haematological investigations conducted at the end of dosing period on day 29 and at the end of recovery period on day 43, revealed following significant changes in the values of different parameters studied when compared with that of respective controls, however the increase/decrease in the values obtained was within normal biological and laboratory limits or the effect was not dose dependent.
Male :
MCHC : Increased levels were observed in animals from 1000 mg/kg reversal dose group, sacrificed on day 43 (p<0.05) and
HCT : Decreased levels were observed in animals from 1000 mg/kg reversal dose group, sacrificed on day 43 (p<0.05).
Female :
Platelets : Increased values were obtained for animals from 250 mg/kg, 500 mg/kg and 1000 mg/kg dose groups, sacrificed on day 29 (p<0.05),Hb, HCT and MCHC : Decreased values were obtained for animals from 250 mg/kg, 500 mg/kg and 1000 mg/kg dose groups, sacrificed on day 29 (p<0.05),
Total RBC : Decreased values were obtained for animals from 500 mg/kg and 1000 mg/kg dose groups, sacrificed on day 29 (p<0.05) and
Total RBC : Decreased levels were observed in animals from 1000 mg/kg reversal dose group, sacrificed on day 43 (p<0.05).
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
2. Bilirubinuria was observed at 24 months in male and female at 1% and females at 3%.
3. Male and Female -
Biochemical investigations conducted at the end of dosing period on day 29 and at the end of recovery period on day 43, revealed following significant changes in the values of different parameters s
tudied when compared with that of respective controls, however the increase/decreased in the valuesobtained was within normal biological and laboratory limits or the effect was not dose dependent.
Male :
Total Bilirubin : Elevated levels were observed in animals from 1000 mg/kg dose group, sacrificed on day 29 (p<0.05),
Creatinine : Elevated levels were observed in animals from 250 mg/kg dose group, sacrificed on day 29 (p<0.05),
Glucose : Decreased levels were observed in animals from 500 mg/kg dose group, sacrificed on day 29 (p<0.05),
Calcium : Decreased levels were observed in animals from 500 mg/kg and 1000 mg/kg dose groups, sacrificed on day 29 (p<0.05)
Globulin and Bile Acid : Decreased levels were observed in animals from 250 mg/kg dose group, sacrificed on day 29 (p<0.05)
Total Cholesterol : Elevated levels were observed in animals from 1000 mg/kg reversal dose group, sacrificed on day 43 (p<0.05)
Glucose : Decreased levels were observed in animals from 1000 mg/kg reversal dose group, sacrificed on day 43 (p<0.05).
Female :
Total Protein and Aspartate Aminotransferase : Elevated levels were observed in animals from 250 mg/kg and 500 mg/kg dose groups, sacrificed on day 29 (p<0.05)Total Bilirubin : Elevated levels were observed in animals from 250 mg/kg, 500 mg/kg and 1000 mg/kg dose groups, sacrificed on day 29 (p<0.05),
Albumin : Elevated levels were observed in animals from 500 mg/kg dose group, sacrificed on day 29(p<0.05),
Calcium : Decreased levels were observed in animals from 250 mg/kg and 500 mg/kg dose groups,sacrificed on day 29 (p<0.05),
Triglycerides : Decreased levels were observed in animals from 250 mg/kg dose group, sacrificed onday 29 (p<0.05) and
Total Protein : Elevated levels were observed in animals from 1000 mg/kg reversal dose group,sacrificed on day 43 (p<0.05).,
Urinalysis findings:
no effects observed
Description (incidence and severity):
3. Total Bilirubin : Elevated levels were observed in animals from 250 mg/kg, 500 mg/kg and 1000 mg/kg dose groups, sacrificed on day 29 (p<0.05),
Albumin : Elevated levels were observed in animals from 500 mg/kg dose group, sacrificed on day 29 (p<0.05),
Calcium : Decreased levels were observed in animals from 250 mg/kg and 500 mg/kg dose groups, sacrificed on day 29 (p<0.05),
Triglycerides : Decreased levels were observed in animals from 250 mg/kg dose group, sacrificed on day 29 (p<0.05) and
Total Protein : Elevated levels were observed in animals from 1000 mg/kg reversal dose group, sacrificed on day 43 (p<0.05).
Behaviour (functional findings):
no effects observed
Description (incidence and severity):
3. Sensory Reactivity Observations: All animals from control and different dose groups showed normal arousal level, visual response, touch response, auditory response, tail pinch response and visual placing response. Normal air righting reflex was observed in all animals from control and different dose groups in week 4.
Grip Strength: Grip strength values observed in male and female animals for control and different dose groups were comparable.
Motor Activity: Motor activity values observed in male and female animals for control and different dose groups were comparable.
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
3. Male -
In comparison with controls at the end of dosing on day 29, organ weight data of animals from 250 mg/kg, 500 mg/kg and 1000 mg/kg dose groups revealed increased relative weights of liver (p<0.05). Increased relative weights of kidneys (p<0.05) were observed in animals from 250 mg/kg dose group.In addition, decreased relative weights of kidneys and heart (p<0.05) were observed in animals from 1000 mg/kg dose group. Decreased relative weights of adrenals (p<0.05) were observed in animals from 250 mg/kg and 1000 mg/kg dose groups. Decreased relative weights of thymus (p<0.05) were observed in animals from 500 mg/kg and 1000 mg/kg dose groups. In comparison with controls at the end of post-dosing recovery period on day 43, organ weight data of animals from 1000 mg/kg reversal group revealed decreased relative weights of kidneys and prostate+ seminal vesicle with coagulation gland as whole (p<0.05).
Female -In comparison with controls at the end of dosing on day 29, organ weight data of animals from 250 mg/kg, 500 mg/kg and 1000 mg/kg dose groups revealed increased relative weights of liver (p<0.05). In comparison with controls at the end of post-dosing recovery period on day 43, organ weight data of animals from 1000 mg/kg reversal group was found to be comparable.Although significant changes in the values of organ weight were observed in male and female animals from different dose groups, no related gross pathological and histopathological findings were
seen, hence these findings were considered to be of no toxicological importance.
Gross pathological findings:
no effects observed
Description (incidence and severity):
2. The test animals sacrificed at 52 weeks did not present any gross changes in the organs or viscera attributable to the test material. Autopsies performed on the animals which died during the second year of the study did not reveal any consistent gross changes.
3. Gross pathological examination revealed test item coloured perianal region externally and test item coloured stomach mucosa in male and female animals from 250 mg/kg, 500 mg/kg and 1000 mg/kg
dose groups.Gross pathological examination in male and female animals from control, control reversal and 1000 mg/kg reversal dose groups did not reveal any abnormality.
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
2. Microscopic findings at two years revealed no compound related effects on the kidneys or other tissues in either sex.
3. No treatment related histopathological changes were evident in male and female animals from control and high dose groups.Incidental, physiological and congenital histopathological changes which were covered in the background historical data of the pathology from control and high dose groups includes minimal, focal to multifocal periportal mononuclear cells infiltration in liver; minimal, multifocal tubular eosinophilic luminal secretion in the kidneys; minimal, multifocal brown pigmentation in spleen; minimal, diffuse dilatation of zona reticularis and/or minimal, multifocal vacuolation in zona fasciculata in the adrenals; minimal, luminal seminal coagulum in urinary bladder; minimal, luminal dilatation in the uterus;minimal, multifocal dilatation of Brunner’s gland in the duodenum; presence of persistent Rathke’s pouch in the pituitary in male and female animals from control and high dose group.
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
behaviour (functional findings)
body weight and weight gain
clinical biochemistry
clinical signs
food consumption and compound intake
gross pathology
haematology
histopathology: non-neoplastic
mortality
Critical effects observed:
not specified
Conclusions:
Based on the available results and applying the weight of evidence approach, the test chemical can be considered to be non-toxic to living organisms when dosed repeatedly via oral route. The No Observed Adverse Effect Level can be considered to be >1000 mg/kgbw/day. Comparing the above annotations with the criteria of CLP Regulation, the test chemical can be classified under the category “Not Classified”.
Executive summary:

Various studies have been reviewed to determine the exact toxic dose of the test chemical when dosed repeatedly to test animals. These include in vivo experimental studies performed on rats for the test chemicals.

Chronic repeated dose oral toxicity study was performed to evaluate the toxic nature of the test chemical. Male and female albino rats were used in the study. The test compound was mixed with diet and used at dose levels of 0, 0.25, 1.0 or 3.0 % (0, 100, 400, 1200 mg/kg/day). The rats were treated for 2 years. The treated animals were noted for clinical signs, mortality, changes in food consumption, hematology, blood chemistry, gross pathology and histopathology. Appearance and behavior of the test rats were generally comparable to those of the controls. No mortality was observed in the treated rats. At the 3% level (1200 mg/Kg/day), food consumption was significant lower than controls for the first six months but comparable to controls during the remainder of the study. Significantly lower hematocrit and hemoglobin values in comparison with controls were obtained at all time intervals except 24 months for the males fed 3%. No such effect was noted in the females. Bilirubinuria was observed at 24 months in male and female at 1% and females at 3%. The test animals sacrificed at 52 weeks did not present any gross changes in the organs or viscera attributable to the test material. Autopsies performed on the animals which died during the second year of the study did not reveal any consistent gross changes. Microscopic findings at two years revealed no compound related effects on the kidneys or other tissues in either sex. Based on the observations made, the No Observed Adverse Effect Level (NOAEL) for the test chemical using male and female Albino rats was considered to be 1200 mg/Kg/day.

 

This is supported by the results of a Repeated Dose 28-day Oral Toxicity study, where Sprague Dawley male and female rats were treated with the test chemical in the concentration of 0, 250 mg/kg, 500 mg/kg and 1000 mg/kg body weight orally by gavage in corn oil. Two additional dose levels were added to the study as control 0 mg/kg (Rev.) and test item 1000 mg/kg (Rev.), in order to study the reversibility or delayed occurrence of symptoms, if any. No effect on survival, clinical sign, body weight, Feed intake, Ophthalmoscopic examination and functional observation battery of treated male and female rats at the end of dosing period of 28 days and the recovery period of 14 days. Similarly, in male animals at the end of the recovery period on day 43,revealed statistically significant increase in the values of MCHC and statistically significant decrease in the values of HCT at 1000 mg/kg and in female on day 29, revealed statistically significant increase in the values of Platelets at 250 mg/kg, 500 mg/kg and 1000 mg/kg, and statistically significant decrease in the values of Hb, HCT and MCHC at 250 mg/kg, 500 mg/kg and 1000 mg/kg, and Total RBC at 500 mg/kg and 1000 mg/kg, Haematological analysis in female animals conducted at the end of the recovery period on day 43, revealed statistically significant decrease in the values of Total RBC at 1000 mg/kg. In male and female animals at the end of the dosing period on day 29, revealed statistically significant increase in the values of Total Bilirubin at 1000 mg/kg and Creatinine at 250 mg/kg, in male, Total Protein and Aspartate Aminotransferase at 250 mg/kg and 500 mg/kg, Total Bilirubin at 250, 500 and 1000 mg/kg, and Albumin at 500 mg/kg in female rat. In addition, statistically significant decrease was observed in the values of Glucose at 500 mg/kg, Calcium at 500 mg/kg and 1000 mg/kg, Globulin and Bile Acid at 250 mg/kg in male rat and, Calcium at 250 and 500 mg/kg, and Triglycerides at 250 mg/kg in female rat. At the end of the recovery period on day 43 (Reversal groups) statistically significant increase were observed in the values of Total Cholesterol at 1000 mg/kg in male rat and Total Protein at 1000 mg/kg in female rat. In addition, statistically significant decrease was observed in the values of Glucose at 1000 mg/kg in male rat. The increase/decrease in the values of various parameters was marginal and within the normal biological and laboratory limits for hematology and clinical chemistry parameters. In Urine analysis conducted during 4th and 6th week of dosing period (on day 23, 24, 25, 26 and 43), revealed no abnormality attributable to the treatment except for higher volume of urine was observed in female animals from 250 mg/kg dose group and considered to be incidental and of no toxicological importance. In addition, at termination of dosing on day 29, male animals from 250, 500 and 1000 mg/kg dose groups revealed increased relative weights of liver when compared with that of controls. In addition, increased relative weights of kidneys were observed in male animals from 250 mg/kg dose group, when compared with that of controls. Decreased relative weights of kidneys and heart weight at 1000 mg/kg in male as compared to controls. In addition, decreased relative weights of adrenals were observed in male animals from 250 and 1000 mg/kg dose groups when compared with that of controls. Decreased relative weights of thymus were observed in male animals from 500 mg/kg and 1000 mg/kg dose groups when compared with that of controls. Organ weight data of male animals sacrificed on day 43 from 1000 mg/kg reversal group, revealed decreased relative weights of kidneys and prostate + seminal vesicle with coagulation gland as whole when compared with that of controls. At termination of dosing on day 29, female animals from 250 mg/kg, 500 mg/kg and 1000 mg/kg dose groups revealed increased relative weights of liver. Organ weight data of female animals sacrificed on day 43 from 1000 mg/kg reversal group, was found to be comparable with that of controls. Although significant changes in the values of organ weight were observed in male and female animals from different dose groups, no related gross pathological and histopathological findings were seen, hence these findings were considered to be of no toxicological importance. Test item coloured perianal region externally and test item coloured stomach mucosa in male and female animals at 250, 500 and 1000 mg/kg dose groups. Gross pathological examination in male and female animals from control, control reversal and 1000 mg/kg reversal dose groups did not reveal any abnormality. Histopathological examination did not reveal any abnormality attributable. Therefore, No Observed Adverse Effect Level (NOAEL) of the test chemical in the Sprague Dawley rat via oral route, over a period of 28 days was found to be 1000 mg/kg body weight in male and female animals.

Based on the available results and applying the weight of evidence approach, the test chemical can be considered to be non-toxic to living organisms when dosed repeatedly via oral route. The No Observed Adverse Effect Level can be considered to be >1000 mg/kgbw/day. Comparing the above annotations with the criteria of CLP Regulation, the test chemical can be classified under the category “Not Classified”.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 200 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
Klimisch Rating 2

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: inhalation
Data waiving:
exposure considerations
Justification for data waiving:
a short-term toxicity study does not need to be conducted because exposure of humans via inhalation in production and/or use is not likely as based on the provided thorough and rigorous exposure assessment
Endpoint conclusion
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:
short-term repeated dose toxicity: dermal
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
a short-term toxicity study does not need to be conducted because exposure of humans via the dermal route in production and/or use is not likely as based on the provided thorough and rigorous exposure assessment
Endpoint conclusion
Quality of whole database:
waiver

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Repeated Dose Toxicity: oral

Various studies have been reviewed to determine the exact toxic dose of the test chemical when dosed repeatedly to test animals. These include in vivo experimental studies performed on rats for the test chemicals.

Chronic repeated dose oral toxicity study was performed to evaluate the toxic nature of the test chemical. Male and female albino rats were used in the study. The test compound was mixed with diet and used at dose levels of 0, 0.25, 1.0 or 3.0 % (0, 100, 400, 1200 mg/kg/day). The rats were treated for 2 years. The treated animals were noted for clinical signs, mortality, changes in food consumption, hematology, blood chemistry, gross pathology and histopathology. Appearance and behavior of the test rats were generally comparable to those of the controls. No mortality was observed in the treated rats. At the 3% level (1200 mg/Kg/day), food consumption was significant lower than controls for the first six months but comparable to controls during the remainder of the study. Significantly lower hematocrit and hemoglobin values in comparison with controls were obtained at all time intervals except 24 months for the males fed 3%. No such effect was noted in the females. Bilirubinuria was observed at 24 months in male and female at 1% and females at 3%. The test animals sacrificed at 52 weeks did not present any gross changes in the organs or viscera attributable to the test material. Autopsies performed on the animals which died during the second year of the study did not reveal any consistent gross changes. Microscopic findings at two years revealed no compound related effects on the kidneys or other tissues in either sex. Based on the observations made, the No Observed Adverse Effect Level (NOAEL) for the test chemical using male and female Albino rats was considered to be 1200 mg/Kg/day.

 

This is supported by the results of a Repeated Dose 28-day Oral Toxicity study, where Sprague Dawley male and female rats were treated with the test chemical in the concentration of 0, 250 mg/kg, 500 mg/kg and 1000 mg/kg body weight orally by gavage in corn oil. Two additional dose levels were added to the study as control 0 mg/kg (Rev.) and test item 1000 mg/kg (Rev.), in order to study the reversibility or delayed occurrence of symptoms, if any. No effect on survival, clinical sign, body weight, Feed intake, Ophthalmoscopic examination and functional observation battery of treated male and female rats at the end of dosing period of 28 days and the recovery period of 14 days. Similarly, in male animals at the end of the recovery period on day 43,revealed statistically significant increase in the values of MCHC and statistically significant decrease in the values of HCT at 1000 mg/kg and in female on day 29, revealed statistically significant increase in the values of Platelets at 250 mg/kg, 500 mg/kg and 1000 mg/kg, and statistically significant decrease in the values of Hb, HCT and MCHC at 250 mg/kg, 500 mg/kg and 1000 mg/kg, and Total RBC at 500 mg/kg and 1000 mg/kg, Haematological analysis in female animals conducted at the end of the recovery period on day 43, revealed statistically significant decrease in the values of Total RBC at 1000 mg/kg. In male and female animals at the end of the dosing period on day 29, revealed statistically significant increase in the values of Total Bilirubin at 1000 mg/kg and Creatinine at 250 mg/kg, in male, Total Protein and Aspartate Aminotransferase at 250 mg/kg and 500 mg/kg, Total Bilirubin at 250, 500 and 1000 mg/kg, and Albumin at 500 mg/kg in female rat. In addition, statistically significant decrease was observed in the values of Glucose at 500 mg/kg, Calcium at 500 mg/kg and 1000 mg/kg, Globulin and Bile Acid at 250 mg/kg in male rat and, Calcium at 250 and 500 mg/kg, and Triglycerides at 250 mg/kg in female rat. At the end of the recovery period on day 43 (Reversal groups) statistically significant increase were observed in the values of Total Cholesterol at 1000 mg/kg in male rat and Total Protein at 1000 mg/kg in female rat. In addition, statistically significant decrease was observed in the values of Glucose at 1000 mg/kg in male rat. The increase/decrease in the values of various parameters was marginal and within the normal biological and laboratory limits for hematology and clinical chemistry parameters. In Urine analysis conducted during 4th and 6th week of dosing period (on day 23, 24, 25, 26 and 43), revealed no abnormality attributable to the treatment except for higher volume of urine was observed in female animals from 250 mg/kg dose group and considered to be incidental and of no toxicological importance. In addition, at termination of dosing on day 29, male animals from 250, 500 and 1000 mg/kg dose groups revealed increased relative weights of liver when compared with that of controls. In addition, increased relative weights of kidneys were observed in male animals from 250 mg/kg dose group, when compared with that of controls. Decreased relative weights of kidneys and heart weight at 1000 mg/kg in male as compared to controls. In addition, decreased relative weights of adrenals were observed in male animals from 250 and 1000 mg/kg dose groups when compared with that of controls. Decreased relative weights of thymus were observed in male animals from 500 mg/kg and 1000 mg/kg dose groups when compared with that of controls. Organ weight data of male animals sacrificed on day 43 from 1000 mg/kg reversal group, revealed decreased relative weights of kidneys and prostate + seminal vesicle with coagulation gland as whole when compared with that of controls. At termination of dosing on day 29, female animals from 250 mg/kg, 500 mg/kg and 1000 mg/kg dose groups revealed increased relative weights of liver. Organ weight data of female animals sacrificed on day 43 from 1000 mg/kg reversal group, was found to be comparable with that of controls. Although significant changes in the values of organ weight were observed in male and female animals from different dose groups, no related gross pathological and histopathological findings were seen, hence these findings were considered to be of no toxicological importance. Test item coloured perianal region externally and test item coloured stomach mucosa in male and female animals at 250, 500 and 1000 mg/kg dose groups. Gross pathological examination in male and female animals from control, control reversal and 1000 mg/kg reversal dose groups did not reveal any abnormality. Histopathological examination did not reveal any abnormality attributable. Therefore, No Observed Adverse Effect Level (NOAEL) of the test chemical in the Sprague Dawley rat via oral route, over a period of 28 days was found to be 1000 mg/kg body weight in male and female animals.

Based on the available results and applying the weight of evidence approach, the test chemical can be considered to be non-toxic to living organisms when dosed repeatedly via oral route. The No Observed Adverse Effect Level can be considered to be >1000 mg/kgbw/day. Comparing the above annotations with the criteria of CLP Regulation, the test chemical can be classified under the category “Not Classified”.

Repeated Dose toxicity: Inhalation

A short-term toxicity study need not be conducted as because exposure of humans via inhalation route in production and/or use is not likely based on the provided thorough and rigorous exposure.The estimated vapour pressure of the test chemical was 5.47E-14 mm Hg. Hence, this endpoint was considered for waiver.

Repeated Dose Toxicity: Dermal

A short-term toxicity study need not be conducted as because exposure of humans via dermal route in production and/or use is not likely based on the provided thorough and rigorous exposure assessment. The acute dermal toxicity value for the test chemical (as provided in section 7.2.3) is >2000 mg/kg body weight. The substance failed to cause any dermal irritation as well sensitization. Considering this, the end point for repeated dermal toxicity is considered as waiver.

Justification for classification or non-classification

Based on the available results, the test chemical is not likely to cause any toxicity to living organisms, when exposed repeatedly via oral, dermal or inhalation route of exposure. Hence, the test chemical can be classified under the category "Not Classified" as per CLP Regulation.