Dodecan-5-olide

Endpoint summary

Description of key information

Repeated dose toxicity: Oral

Based on the data available, the No Observed Adverse Effect Level (NOAEL) for the test chemical was considered to be 1000mg/Kg bw using male and female rats. Hence the test chemical is not likely to be toxic upon repeated exposure by oral route as per the criteria mentioned in CLP regulation.

Repeated dose toxicity: inhalation

The repeated dose  inhalation toxicity study need not be conducted because exposure to humans via inhalation route is not likely taking into account due to the low vapour pressure of the test chemical, which is reported to be 0.00099 mm Hg. Thus, exposure to inhalable dust, mist and vapour of the chemical is highly unlikely. Therefore this study is considered for waiver.

Repeated dose toxicity: dermal

The acute toxicity value for test chemical (as provided in section 7.2.3) is >2000 mg/kg body weight. Also, given the use of the chemical; repeated exposure by the dermal route is unlikely since the use of gloves is common practice in industries. Thus, it is expected that test chemical shall not exhibit 28 day repeated dose toxicity by the dermal route. In addition, there is no data available that suggests that test chemical shall exhibit repeated dose toxicity by the dermal route. Hence this end point was considered for 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:

Experimental data from various test chemicals

Justification for type of information:

Weight of evidence approach based on the available information from various test chemicals.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

Principles of method if other than guideline:

WoE report is based on repeated dose toxicity studies on rats

GLP compliance:

not specified

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

2.TEST ANIMALS
- Source:National Institute of Biosciences
- Females (if applicable) nulliparous and non-pregnant: Yes
- Age at study initiation:6 to 8 weeks old
- Weight at study initiation: Male: 145.6-186.4 g; Female: 124.7-154.0 g
- Fasting period before study:
- Housing: The rats were housed in polycarbonate cages with paddy as bedding.
After allocation to respective dose groups rats were housed 2/sex/cage.
- Diet (e.g. ad libitum): Rodent feed supplied by the Nutrivet Life Sciences 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

DETAILS OF FOOD AND WATER QUALITY:

ENVIRONMENTAL CONDITIONS
- Temperature (°C):19.8 °C to 23.1 °C
- Humidity (%):48.5% to 60.4%
- Air changes (per hr): at least 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 was provided to the room.

IN-LIFE DATES:
From: 25-10-2017
To:16 February 2018
3.TEST ANIMALS
- Source:National Institute of Biosciences
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: (P) x wks; (F1) x wks: 6 to 8 weeks
- Weight at study initiation: (P) Males: x-x g; Females: x-x g; (F1) Males: x-x g; Females: x-x g: Male - 165.53 g (= 100 %) and Female - 150.64 g (= 100 %)
- Fasting period before study:
- Housing: The rats were housed in polycarbonate cages with paddy as bedding.
After allocation to respective dose groups rats were housed 2/sex/cage.
Identified by the picric acid marking. A group of animals in one cage was additionally identified by the label affixed to each cage. A label according to groups identified the cage and each label contained information on cage and study number. It also bear species, strain, sex and identification numbers of rats within it.
- Use of restrainers for preventing ingestion (if dermal): yes/no
- Diet (e.g. ad libitum): Rodent feed, 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

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3°C (actual range, 20.0 °C to 21.9 °C)
- Humidity (%):30% to 70% (actual range, 47.0% to 59.7%).
- Air changes (per hr): at least 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 was provided to the room.

IN-LIFE DATES:
From: 29-11-2017
To:26-2-2018

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

2.PREPARATION OF DOSING SOLUTIONS:The test item was diluted with corn oil for preparation of solution(s).
The solution(s) of test chemical 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.

DIET PREPARATION
- Rate of preparation of diet (frequency):
- Mixing appropriate amounts with (Type of food):
- Storage temperature of food:

VEHICLE
- Justification for use and choice of vehicle (if other than water): corn oil
- Concentration in vehicle: 0, 250, 500 and 1000 mg/kg body weight
- Amount of vehicle (if gavage): 10 ml/kg body weight
- Lot/batch no. (if required):
- Purity:
3.PREPARATION OF DOSING SOLUTIONS: The test item was diluted with corn oil for preparation of solution(s).
The solution(s) of test chemical 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. The concentration of the test item was varied so as to maintain the dose volume constant at or upto 10 ml/kg body weight.

DIET PREPARATION
- Rate of preparation of diet (frequency):
- Mixing appropriate amounts with (Type of food):
- Storage temperature of food:

VEHICLE
- Justification for use and choice of vehicle (if other than water): 0 (vehicle), 250,500 and 1000 mg/kg body weight
- Concentration in vehicle: Corn oil
- Amount of vehicle (if gavage): 10 ml/kg body weight
- Lot/batch no. (if required):
- Purity:

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

concentration and stability were analyzed

Duration of treatment / exposure:

28 days

Frequency of treatment:

Daily

Remarks:

Study 2.
0,250,500,1000mg/kg bw /day
Study 3
0,250,500,1000mg/kg bw /day

No. of animals per sex per dose:

2.Total: 72
0 mg/kg bw/day: 6 male ,6 female
0 mg/kg bw/day (Reversal): 6 male ,6 female
250 mg/kg bw/day: 6 male ,6 female
500 mg/kg bw/day: 6 male ,6 female
1000 mg/kg bw/day: 6 male ,6 female
1000 mg/kg bw/day (Reversal):: 6 male ,6 female
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:

- Dose selection rationale:
- Rationale for animal assignment (if not random): The animals of uniform body weight were selected.
- Rationale for selecting satellite groups: 6 animals /sex /group for reversal group.
- Post-exposure recovery period in satellite groups: 2 week recovery period
- Section schedule rationale (if not random):

Observations and examinations performed and frequency:

2.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 groups 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, Red Blood Corpuscles, Hematocri, Mean Corpuscular Volume, Mean Corpuscular Hemoglobin, Mean Corpuscular Hemoglobin Concentration, Platelets, White Blood Corpuscles, Reticulocytes, Neutrophils, Lymphocytes, Eosinophils, Monocytes, Basophil and Prothrombin time 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, Blood Urea Nitrogen, Urea Nitrogen, Calculated, Alanine Aminotransferase, Aspartate Aminotransferase, Alkaline Phosphatase, Gamma Glutamyl Transferase, Glucose (mg/dL), Calcium , Phosphorous, Albumin, Total Bilirubin, Creatinine, Total Cholesterol, Triglycerides, Globulin, Calculated, Sodium Potassium, Chloride and Bile acid 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.
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 (x 106 /µL), Hematocrit (%), Mean Corpuscular Volume (fL), Mean Corpuscular Hemoglobin (pg), Mean Corpuscular Hemoglobin Concentration (g/dL), Platelets (x 103 /µL), White Blood Corpuscles (x 103 /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
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.
Following tissue samples of organs were subjected to histopathological examination: Adrenals, Aorta, Brain (cerebrum, cerebellum and pons), Caecum, Cervix, Colon, Duodenum, Epididymides, Eyes, Heart, Ileum, Jejunum, Kidneys, Liver, Lungs, Mesenteric Lymphnodes, Muscles - Skeletal muscle, Oesophagus, Ovaries, Pancreas, Pharyngeal Lymphnodes, Pituitary, Prostate, Rectum, Sciatic Nerve, Seminal Vesicles with coagulation gland, Skin with Mammary Gland, Spleen, Spinal Cord (Cervical, mid thoracic and lumbar), Sternum with bone marrow, Stomach, Testes, Thymus, Trachea, Thyroid / Parathyroid, Urinary Bladder, Uterus, Vagina.
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.

Statistics:

2.&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.No clinical signs of toxicity were observed in the animals throughout the dosing period of 28 days and and during the post-dosing recovery period.
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): No clinical signs of toxicity were observed in the animals throughout the dosing period of 28 days (animal nos.25 to 30).
Group IV (500 mg/kg): No clinical signs of toxicity were observed in the animals throughout the dosing period of 28 days (animal nos.37 to 42).
Group V (1000 mg/kg): No clinical signs of toxicity were observed in the animals throughout the dosing period of 28 days (animal nos.49 to 54).
Group VI (1000 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.61 to 66).

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): No clinical signs of toxicity were observed in the animals throughout the dosing period of 28 days (animal nos.31 to 36).
Group IV (500 mg/kg): No clinical signs of toxicity were observed in the animals throughout the dosing period of 28 days (animal nos.43 to 48).
Group V (1000 mg/kg): No clinical signs of toxicity were observed in the animals throughout the dosing period of 28 days (animal nos.55 to 60).
Group VI (1000 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.67 to 72).

Mortality:

no mortality observed

Description (incidence):

2.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.
3.Male and Female -
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.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

2.Male -
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.

Female -
Animals from control and different dose groups exhibited normal body weight gain throughout the dosing period of 28 days.
Body weight gain of 8.45% above that of control was observed in female animals from 1000 mg/kg reversal group. This effect was not considered to be of any toxicological importance.
3.Male and 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.& 3Animals 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):

2.& 3.No ocular abnormalities were observed on ophthalmological examination in the animals during pre-exposure and at the end of the respective termination.

Haematological findings:

effects observed, non-treatment-related

Description (incidence and severity):

2.Male :
MCHC : Increased levels were observed in animals from 1000 mg/kg reversal dose group, sacrificed on day 43 (p<0.05).
Female :
MCV and MCH : Increased values were obtained for animals from 500 mg/kg dose group, sacrificed on day 29 (p<0.05) and
Total RBC and HCT : Decreased values were obtained for animals from 500 mg/kg dose group, sacrificed on day 29 (p<0.05).

However, the increase/decrease in the values obtained was within normal biological and laboratory limits or the effect was not dose dependent.
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 no significant changes in the values of different parameters studied when compared with that of respective controls.

Clinical biochemistry findings:

effects observed, non-treatment-related

Description (incidence and severity):

2.Male :
Aspartate Aminotransferase, Calcium and Gamma Glutamyl Transferase : Decreased levels were observed in animals from 250 mg/kg dose group, sacrificed on day 29 (p<0.05),
Total Protein, Alanine Aminotransferase and Aspartate Aminotransferase : Decreased levels were observed in animals from 1000 mg/kg dose group, sacrificed on day 29 (p<0.05) and
Calcium : Decreased levels were observed in animals from 500 mg/kg dose group, sacrificed on day 29 (p<0.05).

Female :
Total Bilirubin : Elevated levels were observed in animals from 250 mg/kg dose group, sacrificed on day 29 (p<0.05),
Gamma Glutamyl Transferase : Decreased levels were observed in animals from 250 mg/kg dose group, sacrificed on day 29 (p<0.05),
Calcium and Triglycerides : Decreased 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),
Sodium and Chloride : Elevated levels were observed in animals from 1000 mg/kg reversal dose group, sacrificed on day 43 (p<0.05) and
Total Protein, Globulin and Potassium : Decreased levels were observed in animals from 1000 mg/kg reversal dose group, sacrificed on day 43 (p<0.05).
However the increase/decreased in the values obtained was within normal biological and laboratory limits or the effect was not dose dependent.
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 studied when compared with that of respective controls, however the increase/decreased in the values obtained was within normal biological and laboratory limits or the effect was not dose dependent.
Male :
Chloride : Elevated levels were observed in animals from 500 mg/kg dose group, sacrificed on day 29 (p<0.05),
Total Protein : Decreased levels were observed in animals from 500 mg/kg and 1000 mg/kg dose groups, 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),
Globulin and Sodium : Decreased levels were observed in animals from 500 mg/kg dose group, sacrificed on day 29 (p<0.05) and
Creatinine : Decreased levels were observed in animals from 1000 mg/kg reversal dose group, sacrificed on day 43 (p<0.05).

Female :
Blood Urea Nitrogen and Urea Nitrogen : Elevated levels were observed in animals from 250 mg/kg dose group, sacrificed on day 29 (p<0.05),
Phosphorous : Elevated levels were observed in animals from 250 mg/kg and 1000 mg/kg dose groups, sacrificed on day 29 (p<0.05),
Gamma Glutamyl Transferase and Creatinine : Elevated levels were observed in animals from 250 mg/kg and 500 mg/kg dose groups, sacrificed on day 29 (p<0.05),
Total Protein, Total Bilirubin, Albumin and Globulin: Decreased levels were observed in animals from 500 mg/kg dose group, sacrificed on day 29 (p<0.05),
Alanine Aminotransferase and Alkaline Phosphatase : Decreased levels were observed in animals from 250 mg/kg and 500 mg/kg dose groups, sacrificed on day 29 (p<0.05) and
Alanine Aminotransferase and Calcium : Decreased 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):

2.& 3.No statistically significant variation was observed in the urine analyses conducted at the end of the dosing period in week 4 and 6 (on day 23, 24, 25 and 43) in male and female animals of different dose groups as compared to control group animals, except for higher volume of urine was observed in female animals from 1000 mg/kg reversal dose group (p<0.05). This higher volume of urine analyses were considered to be incidental and of no toxicological importance.

Behaviour (functional findings):

no effects observed

Description (incidence and severity):

2.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.
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 examined

Organ weight findings including organ / body weight ratios:

effects observed, non-treatment-related

Description (incidence and severity):

2.Male -
In comparison with controls at the end of dosing on day 29, male animals from 500 mg/kg dose group revealed increased relative weights of liver. In addition, decreased relative weights of spleen were observed in male animals from 250 mg/kg and 500 mg/kg dose groups when compared with that of controls (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.

Female -
In comparison with controls at the end of dosing on day 29, organ weight data of animals from 250 mg/kg dose group revealed increased relative weights of liver, kidneys and heart (p<0.05). Increased relative weights of liver, kidneys and uterus (p<0.05) were observed in animals from 500 mg/kg dose group. In addition, decreased relative weights of ovaries (p<0.05) were observed in animals from 500 mg/kg and 1000 mg/kg dose groups. Decreased relative weights of adrenals (p<0.05) were observed in animals from 1000 mg/kg dose group.
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 increased relative weights of ovaries (p<0.05).

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.
3.Male -
In comparison with controls at the end of dosing on day 29, male animals from 1000 mg/kg dose group revealed decreased relative weights of kidneys and adrenals (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 revealed increased relative weights of liver, kidneys and spleen (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 dose group revealed increased relative weights of kidneys (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 revealed increased relative weights of liver, kidneys, heart and spleen (p<0.05).

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&3.Gross pathological examination on male and female animals from control and different dose groups did not reveal any abnormality.

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

effects observed, non-treatment-related

Description (incidence and severity):

2&3.No treatment related histopathological changes were evident in male and female animals from control and high dose groups.
Incidental and physiological 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, focal mononuclear cells infiltration and/or tubular dilatation in the kidneys; minimal, focal to multifocal increase 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, diffuse luminal dilatation in the uterus 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 (actual dose received)

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

neuropathology

ophthalmological examination

organ weights and organ / body weight ratios

urinalysis

Remarks on result:

other: No effect observed

Critical effects observed:

no

Conclusions:

No Observed Adverse Effect Level (NOAEL) of test chemical in the Sprague Dawley rat via oral route, over a period of 28 days was considered to be 1000 mg/kg body weight in male and female animals.

Executive summary:

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

Study 2.

In a Repeated Dose 28-day Oral Toxicity study, male and female Sprague Dawley rats were treated wtih test chemical in the concnetration 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 were noted at the end of dosing period of 28 days and the recovery period of 14 days. Similarly, at the end of the dosing period in male rat on day 29, revealed no statistically significant changes in the values of various parameters. Statistically significant increase in the values of MCHC at 1000 mg/kg were observed at the end of the recovery period. Statistically significant increase in the values of MCV and MCH and statistically significant decrease in the values of Total RBC and HCT at 500 mg/kg in female rats. No statistically significant changes in the values of various parameters at the end of the recovery period on day 43 in female rats. Statistically significant increase in the values of Total Bilirubin at 250 mg/kg in female rats, Statistically significant decrease was observed in the values of Aspartate Aminotransferase, Calcium and Gamma Glutamyl Transferase at 250 mg/kg in male rats as well as in Total Protein, Alanine Aminotransferase and Aspartate Aminotransferase at 1000 mg/kg in male, Calcium at 500 mg/kg in malerats, Gamma Glutamyl Transferase at 250 mg/kg in female rat, Calcium and Triglycerides at 250, 500 and 1000 mg/kg in female rats. were observed. In male at the end of the recovery period on day 43, no statistically significant changes in the values of various parameters. In female rats, statistically significant increase was observed in the values of Sodium and Chloride at 1000 mg/kg and statistically significant decrease was observed in the values of Total Protein, Globulin and Potassium at 1000 mg/kg. The increase/decrease in the values of various parameters was marginal and within the normal biological and laboratory limits. No abnormality attributable to the treatment except for higher volume of urine was observed in female animals at 1000 mg/kg reversal dose group and considered to be incidental and of no toxicological importance. Increased relative weights of liver at 500 mg/kg bw, decreased relative weights of spleen at 250 mg/kg and 500 mg/kg were observed in male rat as compared to control. Increased relative weights of liver, kidneys and heart at 250 mg/kg and increased relative weights of liver, kidneys and uterus were observed in female animals at 500 mg/kg as compared to control. Decreased relative weights of ovaries at 500 mg/kg and 1000 mg/kg and decreased relative weights of adrenals at 1000 mg/kg were observed in female rats as compared to control. On day 43 at 1000 mg/kg reversal group, increased relative weights of ovaries were observed as compared to 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. In addition, no Gross pathological and histopathological abnormality were observed in reproductive organ of treated male and femlae rats at the end of the dosing period on day 29 and recovery period on day 43 as compared to control. Therefore, No Observed Adverse Effect Level (NOAEL) of test chemical in the Sprague Dawley rata an oral route, over a period of 28 days was found to be 1000 mg/kg body weight in male and female animals.

Study 3

In a Repeated Dose 28-day Oral Toxicity study, Sprague Dawley male and female rats were treated with 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. No effect on survival, clinical sign, body weight gain, Feed intake, Ophthalmoscopic examination and functional observation battery test of treated male and female rats as compared to control. No statistically significant changes were observed in hematology of treated male and female rats were observed on day 29 and day 43 as compared to control. Statistically significant increase in the values of Chloride at 500 mg/kg, in male and Blood Urea Nitrogen and Urea Nitrogen at 250 mg/kg, Phosphorous at 250 mg/kg and 1000 mg/kg, and Gamma Glutamyl Transferase and Creatinine at 250 mg/kg and 500 mg/kg, in female rat were observed. In addition, statistically significant decrease was observed in the values of Total Protein at 500 mg/kg and 1000 mg/kg, Calcium at 250 mg/kg and 500 mg/kg, Globulin and Sodium at 500 mg/kg, Total Protein, Total Bilirubin, Albumin and Globulin at 500 mg/kg, Alanine Aminotransferase and Alkaline Phosphatase at 250 mg/kg and 500 mg/kg, in female rat were observed. Clinical biochemistry analysis in male and female animals conducted at the end of the recovery period on day 43 (Reversal groups) statistically significant decrease was observed in the values of Creatinine at 1000 mg/kg, in male and Alanine Aminotransferase and Calcium at 1000 mg/kg, in female rats. The increase/decrease in the values of various parameters was marginal and within the normal biological and laboratory limits. No abnormality urine analysis attributable to the treatment except for lower volume of urine was observed in female animals at 1000 mg/kg reversal dose group and considered to be incidental and of no toxicological importance. At termination of dosing on day 29, male rat revealed decreased relative weights of kidneys and adrenals at 1000 mg/kg as compared to controls. Organ weight data of male animals sacrificed on day 43 from 1000 mg/kg reversal group, revealed increased relative weights of liver, kidneys and spleen when compared with that of controls. At termination of dosing on day 29, female animals from 250 mg/kg dose group revealed increased relative weights of kidneys when compared with that of controls. In female rat sacrificed on day 43 at 1000 mg/kg reversal group, revealed increased relative weights of liver, kidneys, heart and spleen as compared to 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. No Gross pathological and Histopathological abnormality attributable to the treatment in male and female rats as compared to control. Therefore, No Observed Adverse Effect Level (NOAEL) of test chemical in the Sprague Dawley rat via oral route, over a period of 28 days was considered to be 1000 mg/kg body weight in male and female animals.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

Data is Klimicsh 2 and from authoritative database

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records

Reference

Endpoint:

repeated dose toxicity: inhalation, other

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

other:

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

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: dermal

Data waiving:

other justification

Justification for data waiving:

other:

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Repeated dose toxicity: Oral

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

Study 2.

In a Repeated Dose 28-day Oral Toxicity study, male and female Sprague Dawley rats were treated wtih test chemical in the concnetration 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 were noted at the end of dosing period of 28 days and the recovery period of 14 days. Similarly, at the end of the dosing period in male rat on day 29, revealed no statistically significant changes in the values of various parameters. Statistically significant increase in the values of MCHC at 1000 mg/kg were observed at the end of the recovery period. Statistically significant increase in the values of MCV and MCH and statistically significant decrease in the values of Total RBC and HCT at 500 mg/kg in female rats. No statistically significant changes in the values of various parameters at the end of the recovery period on day 43 in female rats. Statistically significant increase in the values of Total Bilirubin at 250 mg/kg in female rats, Statistically significant decrease was observed in the values of Aspartate Aminotransferase, Calcium and Gamma Glutamyl Transferase at 250 mg/kg in male rats as well as in Total Protein, Alanine Aminotransferase and Aspartate Aminotransferase at 1000 mg/kg in male, Calcium at 500 mg/kg in malerats, Gamma Glutamyl Transferase at 250 mg/kg in female rat, Calcium and Triglycerides at 250, 500 and 1000 mg/kg in female rats. were observed. In male at the end of the recovery period on day 43, no statistically significant changes in the values of various parameters. In female rats, statistically significant increase was observed in the values of Sodium and Chloride at 1000 mg/kg and statistically significant decrease was observed in the values of Total Protein, Globulin and Potassium at 1000 mg/kg. The increase/decrease in the values of various parameters was marginal and within the normal biological and laboratory limits. No abnormality attributable to the treatment except for higher volume of urine was observed in female animals at 1000 mg/kg reversal dose group and considered to be incidental and of no toxicological importance. Increased relative weights of liver at 500 mg/kg bw, decreased relative weights of spleen at 250 mg/kg and 500 mg/kg were observed in male rat as compared to control. Increased relative weights of liver, kidneys and heart at 250 mg/kg and increased relative weights of liver, kidneys and uterus were observed in female animals at 500 mg/kg as compared to control. Decreased relative weights of ovaries at 500 mg/kg and 1000 mg/kg and decreased relative weights of adrenals at 1000 mg/kg were observed in female rats as compared to control. On day 43 at 1000 mg/kg reversal group, increased relative weights of ovaries were observed as compared to 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. In addition, no Gross pathological and histopathological abnormality were observed in reproductive organ of treated male and femlae rats at the end of the dosing period on day 29 and recovery period on day 43 as compared to control. Therefore, No Observed Adverse Effect Level (NOAEL) of test chemical in the Sprague Dawley rata an oral route, over a period of 28 days was found to be 1000 mg/kg body weight in male and female animals.

Study 3

In a Repeated Dose 28-day Oral Toxicity study, Sprague Dawley male and female rats were treated with 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. No effect on survival, clinical sign, body weight gain, Feed intake, Ophthalmoscopic examination and functional observation battery test of treated male and female rats as compared to control. No statistically significant changes were observed in hematology of treated male and female rats were observed on day 29 and day 43 as compared to control. Statistically significant increase in the values of Chloride at 500 mg/kg, in male and Blood Urea Nitrogen and Urea Nitrogen at 250 mg/kg, Phosphorous at 250 mg/kg and 1000 mg/kg, and Gamma Glutamyl Transferase and Creatinine at 250 mg/kg and 500 mg/kg, in female rat were observed. In addition, statistically significant decrease was observed in the values of Total Protein at 500 mg/kg and 1000 mg/kg, Calcium at 250 mg/kg and 500 mg/kg, Globulin and Sodium at 500 mg/kg, Total Protein, Total Bilirubin, Albumin and Globulin at 500 mg/kg, Alanine Aminotransferase and Alkaline Phosphatase at 250 mg/kg and 500 mg/kg, in female rat were observed. Clinical biochemistry analysis in male and female animals conducted at the end of the recovery period on day 43 (Reversal groups) statistically significant decrease was observed in the values of Creatinine at 1000 mg/kg, in male and Alanine Aminotransferase and Calcium at 1000 mg/kg, in female rats. The increase/decrease in the values of various parameters was marginal and within the normal biological and laboratory limits. No abnormality urine analysis attributable to the treatment except for lower volume of urine was observed in female animals at 1000 mg/kg reversal dose group and considered to be incidental and of no toxicological importance. At termination of dosing on day 29, male rat revealed decreased relative weights of kidneys and adrenals at 1000 mg/kg as compared to controls. Organ weight data of male animals sacrificed on day 43 from 1000 mg/kg reversal group, revealed increased relative weights of liver, kidneys and spleen when compared with that of controls. At termination of dosing on day 29, female animals from 250 mg/kg dose group revealed increased relative weights of kidneys when compared with that of controls. In female rat sacrificed on day 43 at 1000 mg/kg reversal group, revealed increased relative weights of liver, kidneys, heart and spleen as compared to 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. No Gross pathological and Histopathological abnormality attributable to the treatment in male and female rats as compared to control. Therefore, No Observed Adverse Effect Level (NOAEL) of test chemical in the Sprague Dawley rat via oral route, over a period of 28 days was considered to be 1000 mg/kg body weight in male and female animals.

Repeated dose toxicity: inhalation

The repeated dose  inhalation toxicity study need not be conducted because exposure to humans via inhalation route is not likely taking into account due to the low vapour pressure of the test chemical, which is reported to be 0.00099 mm Hg. Thus, exposure to inhalable dust, mist and vapour of the chemical is highly unlikely. Therefore this study is considered for waiver.

Repeated dose toxicity: dermal

The acute toxicity value for test chemical (as provided in section 7.2.3) is >2000 mg/kg body weight. Also, given the use of the chemical; repeated exposure by the dermal route is unlikely since the use of gloves is common practice in industries. Thus, it is expected that test chemical shall not exhibit 28 day repeated dose toxicity by the dermal route. In addition, there is no data available that suggests that test chemical shall exhibit repeated dose toxicity by the dermal route. Hence this end point was considered for waiver.

Based on the data available and applying the weight of approach, the test chemical is not likely to be toxic upon repeated exposure by oral, dermal and inhalation route of exposure and hence it is not likely to classify as a toxicant as per the criteria mentioned in CLP regulation.

Justification for classification or non-classification

Based on the data available and applying the weight of approach, the test chemical is not likely to be toxic upon repeated exposure by oral, dermal and inhalation route of exposure and hence it is not likely to classify as a toxicant as per the criteria mentioned in CLP regulation.