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Description of key information

Repeated Dose (Oral) Toxicity Study:

A repeated dose 28-day oral toxicity study was performed to assess the toxicity profile of the test chemical when administered to Sprague -Dawley rats for 28 consecutive days. The test substance was administered via gavage to 6 rats/sex at the dose level of 250, 500 and 1000 mg/kg. Control animals received corn oil as vehicle. Rats were observed for general appearance and clinical signs daily, detailed clinical observation including home cage, handling and open field observations and behavioural assessments were performed weekly. Grips strength, reactivity to sensory stimuli and motor activity were assessed for three consecutive times during the last week of treatment. After completion of 28-day administration period, all rats were sacrificed and examined for gross lesion. At termination day (day 29th), blood samples were collected from all rats for haematology and clinical biochemistry. Vital organs together with male and female reproductive organs were collected, weighted and preserved for histopathological examination. Tissue samples of control and high dose groups were used for histopathological analyses. No case of death was observed during the 28-day administration period. Animals from control and different dose groups exhibited normal body weight gain and feed consumption throughout the dosing period. Daily clinical observation as well as the detailed clinical observations did not reveal any sign of toxicity in dosed rats during the administration period. Functional battery including the assessment of sensory reactivity to various stimuli, grip strength and the motor activity performed during the 4thweek of treatment did not reveal any abnormalities from treated groups when compared to controls. In addition, no treatment related and/or dose dependent changes were noted in haematological and clinical biochemistry parameters up to 1000 mg/kg body weight. External and visceral examination of all treated and control rats did not reveal any abnormality of pathological significance. The absolute and relative organ weights of brain, liver, kidney adrenals, testes, prostate with seminal vesicle and coagulation gland, epididymis, ovaries, uterus, heart, spleen, lung and thymus of dosed rats of either sex did not show treatment-related effect when compared to respective control groups. The relative weight of liver, lung and ovaries of female rats increased significantly at 1000 mg/kg but no related gross pathological, histopathological or haematological changes were seen thus, they were considered incidental alterations with no toxicological importance. In conclusion, the repeated oral exposure of the test chemical exerted no toxicologically significant changes in the morphology and function of a tissue/organ or have produced serious changes to the biochemistry or haematology of the organism when it was orally administered to male and female SD rats for 28 consecutive days.

Repeated Dose (Dermal) Toxicity Study:

A short-term dermal toxicity study does not need to be conducted because exposure of humans via dermal route in production and/or use is unlikely based on the provided thorough and rigorous exposure assessment. Also, the acute dermal toxicity value for test chemical (as provided in section 7.2.3) is >2000 mg/kg body weight. Given the use of the chemical; repeated exposure by the dermal route is unlikely since the use of gloves is common practice in industries. Therefore, it is expected that test chemical shall not exhibit toxicity via dermal route following exposure for 28 days. In addition, there is no data available that suggests that test chemical shall exhibit repeated dose toxicity by the dermal route and therefore this end point was considered for waiver.

Repeated Dose (Inhalation) Toxicity:

A short-term inhalation 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. Also, the given test chemical has very low vapor pressure, < 0.1333 Pa at 20°C, so the potential for the generation of inhalable vapor is very low. The normal conditions of use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalation route will be highly unlikely and therefore the end point of repeated inhalation toxicity 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

Referenceopen allclose all

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Justification for type of information:
Weight of evidence approach based on the available data of the read-across chemicals.
Reason / purpose for cross-reference:
read-across source
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:
According to OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents)
GLP compliance:
not specified
Species:
other: Study 2, 3 and 4: rat
Strain:
other: Study 2: Sprague-Dawley; Study 3: Fischer 344/N; Study 4: Wistar
Details on species / strain selection:
No Data Available
Sex:
male/female
Details on test animals or test system and environmental conditions:
Study 2:
Details on test animal
TEST ANIMALS
- Source: National Institute of Biosciences, Pune.
- Age at study initiation: 6 to 8 weeks old
- Weight at study initiation:
Male 169.50 gg
Female 152.35 g
- Fasting period before study: No data available
- Housing: Animals were housed in polycarbonate cages. Three rats of same sex were housed together in each cage of size 39 cm X 28 cm X 14 cm. Paddy husk was used as bedding material.
- Diet (e.g. ad libitum): Rodent feed supplied by the Nutrivet Life Sciences, Pune, was provided ad libitum from individual feeders on cage top.
- Water (e.g. ad libitum): Water was provided ad libitum from individual bottles attached to the cages. Water was from a local source and passed through the reverse osmosis membrane before use.
- Acclimation period: 5 days prior to dosing.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3°C (actual range: 19.5 °C to 22.8 °C)
- Humidity (%):30% to 70% (actual range: 53.3% to 58.1%).
- Air changes (per hr): Ten air changes per hour of 100% fresh air that has been passed through the HEPA filters.
- Photoperiod (hrs dark / hrs light): An artificial light and dark cycle of 12

Study 3:
- Source: Simonsen Laboratories, Inc. (Gilroy, CA).
- Age at study initiation: 41 days
- Weight at study initiation: Not available
- Fasting period before study: Not available
- Housing: 5 animals per cage in polycarbonate cages with hardwood chips and spun-bonded polyester.
- Diet (e.g. ad libitum): NIH-07 open formula meal rat diet, available ad libitum
- Water (e.g. ad libitum): Automatic watering system, available ad libitum
- Acclimation period: 12 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.9 °± 0.4°C
- Humidity (%): 45.4 to 54%
- Air changes (per hr): minimum of 10 changes/hour
- Photoperiod (hrs dark / hrs light): 12 hours/day

Study 4:
Source: In-House Bred at sa-FORD, Animal Facility (CPCSEA Registration No. 1256/bc/09/CPCSEA)
Health Status: Healthy young adult animals were used for the study. Females were nulliparous and non-pregnant.
Body weight of animals: Male: Minimum: 240 g; Maximum: 315 g
Female: Minimum: 210 g; Maximum: 260 g (Individual body weights were within ± 20% of mean body weight, prior to treatment)
Age: 12 - 13 weeks at the start of Oestrous Cycle evaluation.
Acclimatisation: Animals were acclimatised to the test conditions for 20 days prior to test item administration
Housing: Before the animals are brought in, the study room and cages were cleaned and disinfected. During the study, the floor of the experimental room and work tops were swept and mopped with disinfectant solution every day or as on requirement. Cages were cleaned at regular intervals.
A total 2-3 rats/sex were housed in Polycarbonate cages (size 37 [cm] x 21 [cm], height 20 [cm]). Cage rotation was carried out weekly during study period except during mating for males and females both and during gestation and lactation for females. Sterilized corn cob produced from pure corn, dried and free from dust, procured from approved supplier, was used as bedding material. It was renewed as often as necessary to keep the animals dry and clean.
Bedding material of batch No. SPAR-30/2015 (Sparconn Life Sciences Bangalore) was used in this study and a copy of report of microbial and chemical contaminants analysed periodically by manufacturer of bedding material are incorporated in the raw data.

Environmental conditions:
The room temperature was maintained at 18.30 to 22.70 °C and the relative humidity was kept between 43.90 to 67.60%. Artificial light was set to give a cycle of 12 hours light and 12 hours dark. Air changes were about minimum 12 times per hour and filtered adequately.

Diet :A conventional laboratory pelleted diet of batch no. 004915, 041215 and 041015 from approved supplier (Nutrivet Life Sciences, Pune) was offered ad libitum. The copy of composition, microbial and chemical contaminant reports analysed periodically by manufacturer are incorporated in the raw data.

Water : Aqua guard filtered drinking water in bottles was offered ad libitum. Samples of the drinking water was subjected periodically to bacteriological tests and to chemical contaminant analysis. The latest test results are included in the raw data.
Route of administration:
other: Study 2: oral: gavage; Study 3: oral: feed; study 4: oral: gavage
Vehicle:
other: Study 2: Corn oil; Study 3: NIH-07 open formula meal rat diet: Study 4: Corn oil
Details on oral exposure:
Study 2:
Details on oral exposure
PREPARATION OF DOSING SOLUTIONS: The test chemical was diluted with Corn oil for preparation of dosing solution(s).

DIET PREPARATION
- Rate of preparation of diet (frequency): No data available
- Mixing appropriate amounts with (Type of food): No data available
- Storage temperature of food: No data available

VEHICLE
- Justification for use and choice of vehicle (if other than water): No data available
- Concentration in vehicle: 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 bw/day.
- Amount of vehicle (if gavage): 0.00 mg/ml/day, 26.60 mg/ml/day, 52.38 mg/ml/ day and 105.27 mg/ml/day.
- Lot/batch no. (if required): No data available
- Purity: No data available

Study 3:
DIET PREPARATION
- Rate of preparation of diet (frequency): Doses were prepared weekly
- Mixing appropriate amounts with (Type of food): A premix of feed and the test chemical was prepared, then remaining feed was blended into the premix in a Patterson- Kelly twin-shell blender with the intensifier bar on for 5 minutes and off for 10 minutes.
- Storage temperature of food: The dose formulations were stored at -20°C in sealed, double plastic bags for no longer than 13 days.

Study 4:
PREPARATION OF DOSING SOLUTIONS: The test item was weighed and dissolved in a vehicle (corn oil) to achieve desired concentration of test item. Dose formulation was freshly prepared daily. At the time of dosing, dose formulation was kept on the magnetic stirrer to maintain the homogeneity of test item. The details of dose formulation preparation is maintained in raw data.

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 was used as a vehicle based on the solubility testing
- Concentration in vehicle: 0, 308, 556 and 1000 mg/kg bw
- Amount of vehicle (if gavage): 0.5 ml/kg
- Lot/batch no. (if required): MR301015, MR161215
- Purity:
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Study 2: Analysis for concentration and stability of the test chemical were conducted at Subcontracted Laboratory. Test item formulation samples prepared day 1 (pre-dosing) were sent to Subcontracted Laboratory.

Study 4: Homogeneity and Stability of dose formulation by analysing the sample at different time points (Stability was determined by sampling and analyzing the aliquots from the sample stored at 25 ± 2°C at the time points of 0, 2 and 6 hours).Two replications was analyzed at each time point.The dose formulation analysis were carried out at the start (on the day 1) of treatment, on day 21 and day 40 during the study period.
Duration of treatment / exposure:
Study 2: 28 days consecutively
Study 3: 103 weeks
Study 4:
Male: 47 days
Female : 63 days
Frequency of treatment:
Study 2: Once daily
Study 3: Daily
Study 4: Daily
Remarks:
Study 2: Doses / Concentrations:
0 (vehicle), 250, 500 and 1000 mg/kg bw/day.
Basis: actual ingested

Study 3:
Doses / Concentrations:
0, 3000, 6000 or 12000 ppm (0, 150, 300, 600 mg/kg body weight)

Study 4:
0, 338, 556 and 1000 mg/kg bw/day
No. of animals per sex per dose:
Study 2:
Control: 6 male, 6 female
250 mg/kg bw/day: 6 male, 6 female
500 mg/kg bw/day: 6 male, 6 female
1000mg/kg bw/day: 6 male, 6 female

Study 4:
Total: 124
0 mg/kg bw: 13 male, 13 female
308 mg/kg bwm: 13 male, 13 female
556 mg/kg bwm: 13 male, 13 female
1000 mg/kg bwm: 13 male, 13 female
Control recovery: 5 male, 5 female
1000 mg/kg bw recovery: 5 male, 5 female
Control animals:
yes, concurrent vehicle
Details on study design:
Study 2:
Details on study design
- Dose selection rationale: Based on results from a preliminary 14-day study there was no chenge in the survivel, body weight, Daily clinical observations and Gross pathological examination of 1000 mg/kg/bw/day group. Based on these results, the 28 day study dose levels were finalized as 0 mg/kg, 250 mg/kg, 500 mg/kg and 1000 mg/kg body weight.
- Rationale for animal assignment (if not random): Animals were randomized by sex and body weight
- Rationale for selecting satellite groups: No data available
- Post-exposure recovery period in satellite groups: No data available
- Section schedule rationale (if not random): No data available

Study 3:
- Dose selection rationale: Dietary concentrations selected for the 2-year study in rats were 0, 3000, 6000, or 12000 ppm of the test chemical. The 25000 and 50000 ppm levels were considered too high for males and females because of the reduced mean body weights of and feed consumption by rats receiving 25000 and 50000 ppm and the low survival and microscopic lesions that occurred in 50000 ppm rats in the 13-week study.

Study 4:- Dose selection rationale:
- Rationale for animal assignment (if not random): Vaginal smear of all females was evaluated for regular cyclicity before treatment (14 days). At the time of randomization females not showing regular cycle were euthanised and discarded
- Other: No Data Available
Positive control:
No Data Available
Observations and examinations performed and frequency:
Study 2:
Observations and examinations performed & frequency
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: No data available
- Cage side observations checked in table [No.?] were included. : Rats were observed for Behavior, Alterations, Vocalizations, Respiration and Palpebral closure.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Once daily, At least once a week there after until

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

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): No data available
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data available
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data available

FOOD EFFICIENCY:
- 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: Yes / No / No data

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data available
- Time schedule for examinations: No data available

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: Once a day
- Dose groups that were examined: 0 mg/kg, 250 mg/kg, 500 mg/kg and 1000 mg/kg bw/day.

HAEMATOLOGY: Yes, By using Beckman Coulter haematology analyzer.
- Time schedule for collection of blood: At termination.
- Anaesthetic used for blood collection: No data available
- Animals fasted: Yes, overnight fasted prior to sampling.
- How many animals: Blood collected from all rats of 0 mg/kg, 250 mg/kg, 500 mg/kg and 1000 mg/kg bw/day group at termination.
- Parameters checked in table [No.?] were examined. : Hemoglobin, Red Blood Corpuscles, Hematocrit, Mean Corpuscular Volume, Mean Corpuscular Hemoglobin, Mean Corpuscular Hemoglobin Concentration, Platelets, White Blood Corpuscles, Neutrophils, Lymphocytes, Eosinophils, Monocytes, Basophil and Prothrombin time were checked, Table No.H; Appendix No.VII.

CLINICAL CHEMISTRY: Yes, By using Dimension XpandPlus and Acculyte 5P.
- Time schedule for collection of blood: At termination.
- Animals fasted: Yes, overnight fasted prior to sampling.
- How many animals: Blood collected from all rats of 0 mg/kg, 250 mg/kg, 500 mg/kg and 1000 mg/kg bw/day group at termination.
- Parameters checked in table [No.?] were examined. : Total Protein, Blood Urea Nitrogen, Urea Nitrogen, Alanine Aminotransferase, Aspartate Aminotransferase, Alkaline Phosphatase, Gamma Glutamyl Transferase, Glucose, Calcium, Phosphorous, Albumin, Total Bilirubin, Creatinine , Total Cholesterol, Triglycerides, Globulin Calculated, Sodium, Potassium, Chloride were checked.

URINALYSIS: No data available
- Time schedule for collection of urine: No data available
- Metabolism cages used for collection of urine: No data available
- Animals fasted: No data available
- Parameters checked in table [No.?] were examined. : No data available

NEUROBEHAVIOURAL EXAMINATION: No data available
- Time schedule for examinations: No data available
- Dose groups that were examined: No data available
- Battery of functions tested: sensory activity / grip strength / motor activity / other: Yes

OTHER: Viability, Behavior in Home cage, Vocalizations, Respiration, Palpebral closure, Handling Observations, Urination, Defecation, Prominence of Eye, Lacrimation, Salivation, Piloerection, Examination of Skin / Fur, Stereotype Behaviour, Rearing (Rears), Clonic and Tonic Movements and Severity of Gait were observed.

Study 3:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Observed twice daily.
- Cage side observations checked in table [No.?] were included.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: recorded initially, weekly during first 13 weeks of study, every 4 weeks thereafter, and at end of study

BODY WEIGHT: Yes
- Time schedule for examinations: Animals weighed initially, weekly during first 13 weeks of study, every 4 weeks thereafter, and at end of study.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Feed consumption was measured daily per cage for 5 days once every 4 weeks.

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
- Time schedule for examinations:

OPHTHALMOSCOPIC EXAMINATION: Yes / No / No data
- Time schedule for examinations:
- Dose groups that were examined:

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at 15-month interim evaluations
- Anaesthetic used for blood collection: Yes (carbon dioxide)
- Animals fasted: No data
- How many animals: No data
- Parameters checked: hematocrit, hemoglobin, erythrocytes, mean erythrocyte volume, mean erythrocyte hemoglobin, mean erythrocyte hemoglobin concentration, platelets, reticulocytes, and leukocyte count and differential.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at 15-month interim evaluations
- Animals fasted: No data
- How many animals: No data
- Parameters checked: cholesterol, triglyceride, alkaline phosphatase, creatine kinase, and sorbitol dehydrogenase

URINALYSIS: No
- Time schedule for collection of urine:
- Metabolism cages used for collection of urine: Yes / No / No data
- Animals fasted: Yes / No / No data
- Parameters checked in table [No.?] were examined.

NEUROBEHAVIOURAL EXAMINATION: No
- Time schedule for examinations:
- Dose groups that were examined:
- Battery of functions tested: sensory activity / grip strength / motor activity / other:

OTHER:
Pancreatic enzyme analysis: amylase, lipase and carboxypeptidase.

Study 4:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily (morning and evening)
- Cage side observations checked in table [No.?] were included. : morbidity and mortality were examined.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: once a day, preferably at the same time each day considering the peak period of anticipated effects after dosing.
Detailed clinical examinations were carried out once before the first treatment (to allow for within-subject comparisons) and weekly thereafter.
The detailed clinical examinations were made outside the home cage, at approximately the same time, on each occasion.

BODY WEIGHT: Yes
- Time schedule for examinations: on the first day of dosing, at least weekly thereafter, and at termination. During pregnancy, females were weighed on days 0, 7, 14 and 20 and within 24 hours of parturition (day 0 or 1 post-partum), day 4 post-partum and before terminal sacrifice.

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

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data
- Time schedule for examinations:

OPHTHALMOSCOPIC EXAMINATION: Not specified
- Time schedule for examinations:
- Dose groups that were examined:

HAEMATOLOGY: Yes
- Time schedule for collection of blood: just prior to necropsy.
- Anaesthetic used for blood collection: No
- Animals fasted: Yes, overnight (approximately 16-18 hr) prior to blood collection
- How many animals: five males and five females, randomly selected from each group
- Parameters checked in table [No.?] were examined.: Total Erythrocyte Count (RBC), Hematocrit (HCT), Mean Corpuscular Volume (MCV), Hemoglobin (HGB), Mean Corpuscular Hemoglobin (MCH), Mean Corpuscular Hemoglobin Concentration (MCHC), Platelet Count (PLT), Total Leukocyte count (WBC), Prothombin Time (PT) and Activated Partial Thromboplastin time (aPTT) were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: just prior to necropsy.
- Animals fasted: Yes, overnight (approximately 16-18 hr) prior to blood collection
- How many animals:five males and five females, randomly selected from each group
- Parameters checked in table [No.?] were examined: Glucose (Glu), Cholesterol (Chol), Triglycerides (TRIG), Alanine amino transferase (ALT), Aspartate amino transferase (AST), Calcium, Albumin (Alb), Total Protein (TP), Creatinine (Crea), Phosphorus, Urea, Sodium (Na), Potassium (K), Blood urea nitrogen (BUN) – Calculated, Globulin (Glob) - Calculated, Alb/ Glb (A:G) – Calculated and Bile acidswere examined.

URINALYSIS: Not specified
- Time schedule for collection of urine:
- Metabolism cages used for collection of urine: Not specified
- Animals fasted: Yes / No / Not specified
- Parameters checked in table [No.?] were examined.

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: During the last week of treatment and that of recovery groups, in the last week of recovery period.
- Dose groups that were examined:All dose groups were examined.
- Battery of functions tested: sensory activity / grip strength / motor activity / other:Sensory reactivity to stimuli, assessment of grip strength, hind limb foot splay and motor activity assessment were conducted

IMMUNOLOGY:Not specified
- Time schedule for examinations:
- How many animals:
- Dose groups that were examined:
- Parameters checked in table [No.?] were examined.

OTHER:
Organ weight were examined.
Weighing of brain, adrenals, ovaries with oviduct, testes, epididymides, heart, liver, kidneys, thymus and spleen was performed for randomly selected 5 male and 5 female rats. Testes and epididymides of all male rats were weighed.
Sacrifice and pathology:
Study 2: Sacrifice and pathology
GROSS PATHOLOGY: Yes, Gross necropsy was conducted. All the animals of 0 mg/kg, 250 mg/kg, 500 mg/kg and 1000 mg/kg/bw/day group were sacrificed and gross lesions were noted.

HISTOPATHOLOGY: Yes, Control and treated at the highest dose level of 1000 mg/kg were subjected to sacrifice. Organs examined were Adrenals, Aorta, Brain (cerebrum, cerebellum and pons), Caecum, Cervix, Colon, Duodenum, Epididymides, Eyes, Heart, Ileum, Jejunum, Kidneys, Liver, Lungs, Mesenteric Lymphnodes, Oesophagus, Ovaries, Pancreas, Pituitary gland, Pharyngeal Lymphnodes, Prostate, Rectum, Skeletal Muscles, Skin with Mammary Gland, Spleen, Sternum with bone marrow, Sciatic Nerve, Spinal Cord (Cervical, mid thoracic and lumbar), Stomach, Seminal Vesicles with Coagulation Gland, Testes, Thymus, Thyroid, Trachea, Vagina, Urinary Bladder and Uterus of 1000 mg/kg/bw/day group.

Study 3:
GROSS PATHOLOGY: Yes, Necropsy was performed on all animals. Organs weighed were brain, right kidney, and liver.

HISTOPATHOLOGY: Yes, Complete histopathologic examinations were performed on all animals. In addition to gross lesions, tissue masses, and associated lymph nodes, the tissues examined included: adrenal gland, brain, esophagus, femur (including marrow), heart, kidney, large intestine (cecum, colon, and rectum), liver, lung, mammary gland, nose, ovary, pancreas, parathyroid gland, pituitary gland, preputial or clitoral gland, prostate gland, salivary gland, skeletal muscle, skin, small intestine, spleen, stomach (forestomach and glandular), testis with epididymis and seminal vesicle, thymus, thyroid gland, tongue, trachea, urinary bladder, and uterus.

Study 4:
GROSS PATHOLOGY: Yes, At scheduled sacrifice date, all rats of main and recovery groups were euthanized by over dose of carbon dioxide followed by exsanguination. The animals were examined externally in unopened condition.

HISTOPATHOLOGY: Yes, All the preserved organs (Testes, epididymides, prostate and seminal vesicle with coagulating glands, ovaries, uterus and cervix with vagina) of all the rats, all the preserved tissues of randomly selected five male and five female rats of groups G1 and G4 and preserved thyroid of one male and one female pup of each litter were subjected to histopathological examination. All the tissues were trimmed, processed, embedded in paraffin wax. Sections were cut at a thickness of 3-5 micron and stained with hematoxylin and eosin stain.
Processed tissues were subjected to histopathological examination. The prepared slides were examined under microscope by the Pathologist to note histopathological lesions, if any in different organs. Special attention was paid to observe effect of test item on reproductive system and spermatogenesis.
Other examinations:
No Data Available
Statistics:
Study 2: Data were analyzed for reporting group means and standard deviations with significance between the controls and treated groups, using in-house developed and validated MS-Excel 2003 based statistical software. All the parameters characterized by continuous data such as body weight, per cent body weight change, feed consumption, organ weight, relative organ weight, haematological and clinical chemistry data were subjected to Bartlett’s test to meet the homogeneity of variance before conducting Analyses of Variance (ANOVA) and Dunnett’s t-test. Where the data did not meet the homogeneity of variance, Student’s t-test was performed to calculate significance.
Study 3: The probability of survival was estimated by the product-limit procedure of Kaplan and Meier and is presented in the form of graphs. Animals found dead of other than natural causes were censored from the survival analyses; animals dying from natural causes were not censored. Statistical analyses for possible dose-related effects on survival used Cox’s method for testing two groups for equality and Tarone’s life table test to identify dose-related trends. All reported P values for the survival analyses are two sided.
Study 4: Raw data was analysed using statistical software “Sigma Plot 11.0” (Supplied by Cranes Software International Ltd. Bangalore). The mean and standard deviation was calculated using the software and all data was summarized in tabular form. All continuous data (body weight, feed consumption, Functional Observational Battery parameters, hematology, clinical chemistry, absolute and relative organ weights, maternal and pup parameters etc.) were checked for normality using Shapiro Wilk test. All homogenous data was analysed using ANOVA and data showing significance in their variances was subjected to Dunnett’s t-test. All heterogeneous data was analysed using F test and Student’s t-test, Dunn’s Test, Kruskal-Wallis, ANOVA on ranks,
Clinical signs:
no effects observed
Description (incidence and severity):
Study 2: Daily clinical observations did not reveal any signs of toxicity in male and female animals from of 0 mg/kg, 250 mg/kg, 500 mg/kg and 1000 mg/kg bw/day dose groups.

Study 3: No clinical findings were associated with the administration of the test chemical.

Study 4: No apparent treatment related clinical signs were observed in any of the animals throughout the treatment and recovery period. Detailed clinical examinations like Home cage observation, Handling observation and Open field observation of all animals were observed to be normal during study period. Statistically significant decrease was observed in number of rears of G3 (556 mg/kg body weight) and G4 (1000 mg/kg body weight) male on pre-treatment as compared to control G1 (0 mg/kg body weight). The statistically significant increase was observed in number of urine pools of G3 (556 mg/kg body weight) and G4 (1000 mg/kg body weight) male at pre-treatment as compared to control G1 (0 mg/kg body weight). Statistically significant increase was observed in number of fecal bolus of G3 (556 mg/kg body weight) male at pre-treatment as compared to control G1 (0 mg/kg body weight). Statistically significant increase was observed in number of rears of G4 (1000 mg/kg body weight) male at week 4 as compared to control G1 (0 mg/kg body weight). Statistically significant increase was observed in number of urine pools of G3 (556 mg/kg body weight) male at week 6 as compared to control G1 (0 mg/kg body weight). Statistically significant increase was observed in number of rears of G2 (308 mg/kg body weight), G3 (556 mg/kg body weight) and G4 (1000 mg/kg body weight) female at pre-treatment as compared to control G1 (0 mg/kg body weight). Statistically significant increase was observed in number of fecal bolus of G4 (1000 mg/kg body weight) female at week 5 as compared to control G1 (0 mg/kg body weight). The above changes observed were inconsistent/ biologically insignificant and not dose dependant, hence considered as incidental and not attributed to the effect of test item administration.
Mortality:
no mortality observed
Description (incidence):
Study 2: No mortality were observed in any of the traeted groups of 0 mg/kg, 250 mg/kg, 500 mg/kg and 1000 mg/kg bw/day.

Study 3: The survival rates among exposed rats were similar to those of the controls.

Study 4: No mortality or morbidity was observed in any animal of the control and treatment groups throughout the study period.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Study 2: All the rat of 0 mg/kg, 250 mg/kg, 500 mg/kg and 1000 mg/kg/bw/day dose groups exhibited normal body weight gain at the end of the study period of 28 days.

Study 3: The mean body weights of male rat groups receiving 3000 and 6000 ppm were similar to those of the control group throughout the study. The mean body weights of the 12000 ppm males and exposed females approximately 5% lower than those of the control groups.

Study 4: A statistically significant decrease was observed in body weight of G4 (1000 mg/kg body weight) male on day 30 as compared to control G1 (0 mg/kg body weight). Statistically significant decrease was observed in body weight of G4 (1000 mg/kg body weight) female on day 20 of gestation as compared to control G1 (0 mg/kg body weight). Statistically significant decrease was observed in body weight of G4-R (1000 mg/kg body weight) male on day 29, 36, 41 as compared to control G1-R (0 mg/kg body weight). Statistically significant decrease was observed in percent body weight change of G3 (556 mg/kg body weight) and G4 (1000 mg/kg body weight) male on day 1-8, 1-14 whereas statistically significant decrease was observed in percent body weight change of G4 (1000 mg/kg body weight) male on day 1-21, 1-28, 1-30, 1-37, 1-44, 1-46 as compared to control G1-R (0 mg/kg body weight). Statistically significant decrease was observed in percent body weight change during gestation period of G4 (1000 mg/kg body weight) female on day 0-14, 0-20 as compared to control G1 (0 mg/kg body weight). Statistically significant decrease was observed in percent body weight change of G4-R (1000 mg/kg body weight) male on day 1-8, 1-15, 1-22, 1-29 as compared to control G1-R (0 mg/kg body weight).
Body weight and Percent body weight changes in animals of the all other test groups of both the sexes was comparable and did not show any significant difference as compared to the respective control group. These changes observed were inconsistent, hence not considered as effect of the test item administration.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
Study 2: Food consumption and compound intake Food intake of animals for control and 250 mg/kg, 500 mg/kg and 1000 mg/kg /bw/day dose groups were found to be normale throughout the study period of 28 days.

Study 3: The average feed consumption by all exposure groups was similar to that of the control groups, with only the 12000 ppm males consuming slightly less feed throughout the study.

Study 4: Statistically significant decrease in feed consumption was observed in G4 (1000 mg/kg body weight) female on gestation day 14-20 as compared to the control group G1. Feed consumption in animals of the all other test groups of both the sexes was comparable and did not show any significant difference as compared to the respective control group. Changes observed in feed consumption were inconsistent, hence not considered as effect of the test item administration.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Study 2: Increase in the values of Hb of male rats dosed at 500 mg/kg and 1000 mg/kg, MCHC of male rats dosed at 500 mg/kg, Total WBC of male rats dosed at 1000 mg/kg were observed. Plateles of female rats dosed at 1000 mg/kg were also increased. The increase in the values of different parameters is within the normal laboratory limits.

Study 3: The hematology profiles of rats at the 15-month interim evaluation showed no test chemical-related effects for the parameters measured.

Study 4: All hematological parameters in animals of different treated groups of both the sexes were comparable to their respective control groups, except statistically significant decrease observed for MCHC, WBC in males of G4 (1000 mg/kg body weight) as compared to G1, statistically significant increase observed for aPTT in males of G4 (1000 mg/kg body weight) and G3 (556 mg/kg body weight) as compared to G1. Statistically significant decrease observed for RBC, HCT, HGB, WBC in males of G4-R (1000 mg/kg body weight) as compared to G1-R. Statistically significant decrease observed for PT in females of G3 (556 mg/kg body weight) as compared to G1. Statistically significant decrease observed for MCHC and statistically significant increase observed for RBC, HCT, HGB in females of G4-R (1000 mg/kg body weight) as compared to G1-R. The above changes were inconsistent, not related to the test item and may be due to the preanalytical and analytical variables (J. Robinson and G.O.Evans, 2005).
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
Study 2: Clinical chemistry Increase level of Sodium in male rats dosed at 250 mg/kg and 1000 mg/kg of the test chemical. Chloride levels significantly increase in male rats dosed at 250 mg/kg of the test chemical. Statistically significant increase of Calcium levels in female rats dosed at 250 mg/kg and increase Bilirubin levels in female rats dosed at 500 mg/kg of the test chemical were found well increase level of Sodium in female rats dosed at 250 mg/kg, 500 mg/kg and 1000 mg/kg were observed. Statistically significant decrease of Alkaline Phosphatase levels in female rats dosed at 250 mg/kg, decrease level of Potassium in female rats dosed at 250 mg/kg and 1000 mg/kg and decrease Chloride levels in female rats dosed at 1000 mg/kg of the test chemical were observed. Although there was an increase/decrease in the values the deviations were within the range of normal laboratory limits.

Study 3:
Alkaline phosphatase activity was slightly higher in 12000 ppm males than in control males, but the relationship to chemical administration is inconclusive. Pancreatic enzyme activities were measured in male rats only but no statistically significant differences occurred between any exposure group and the control group for any of the enzymes.

Study 4:
All clinical chemistry parameters in animals of different treated groups of both the sexes were comparable to their respective control groups, except statistically significant increase observed for ALT and statistically significant decrease observed for Sodium (Na) in males of G4 (1000 mg/kg Body weight) as compared to G1. Statistically significant increase observed for Creatinine in males of G2 (308 mg/kg Body weight) as compared to G1. Statistically significant decrease observed for Total Protein and statistically significant increase observed for A/G ratio in females of G3 (556 mg/kg Body weight) as compared to G1. The above changes were inconsistent, not dose dependent hence considered as incidental in nature.
Urinalysis findings:
not specified
Behaviour (functional findings):
effects observed, non-treatment-related
Description (incidence and severity):
Study 4:
The sensory reactivity measurements were comparable and no changes were revealed in any of the animals of all treated groups in both the sexes.
Foot splay and fore limb and hind limb grip strength parameters were comparable and no treatment related changes were revealed in any of the animals of all treated groups except a statistically significant decrease was observed in hindlimb foot splay in G4-R (1000 mg/kg body weight) male as compared to the repective control group G1-R. The above changes observed were inconsistent/ biologically insignificant and not dose dependant, hence, considered as incidental and not attributed to the effect of test item administration. Motor activity measurements were comparable and no changes were revealed in any of the animals from all treated groups of both the sexes as compare to control group except statistically significant decrease was observed in ST=Stereotypic time in G2, G3 and G4 male as compared to control group G1 and G4-R in female as compared to G1-R. The above changes observed were inconsistent, hence considered as incidental and not attributed to the effect of test item administration.
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Description (incidence and severity):
Study 2: Organ weight of 1000 mg/kg/bw/day dose group male animals sacrificed on day 29, was found to be comparable with that of controls. Organ weight of female animals sacrificed on day 29, revealed increased relative weights of liver, ovaries and lungs of 1000 mg/kg dose group.The significant changes in organ weights were observed in female animals from high dose group, the effects are not related to the test chemical.

Study 3: no effects observed

Study 4: At the end of treatment and recovery period, absolute and relative weight of organs of treated rats of either sex did not differ significantly except a significant increase in relative wieght of Adrenal of G4-R male group when compared to the respective control group rats.
Gross pathological findings:
no effects observed
Description (incidence and severity):
Study 2: There were no gross pathological changes in the control and all the treatment groups due to the administration of the test chemical.

Study 3: no effects observed

Study 4: External examination of all male and female rats of control and all treated groups including recovery groups did not reveal any abnormality of pathological significance
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
Study 2: Minimal focal to multifocal periportal mononuclear cell infiltration in the liver; minimal interstitial haemorrhages in the kidneys; minimal alveolar haemorrhages and/or alveolar histiocytosis in the lungs; minimal multifocal haemosiderosis and/or diffused congestion in spleen; minimal eosinophilic infiltration and/or luminal dilatation in uterus; minimal luminal seminal coagulum in the urinary bladder; minimal dilatation of zona reticularis and/or presence of accessory adrenocortical tissue in adrenals; minimal multifocal haemorrhages in thymus; presence of ultimobranchial cysts in thyroid; in male or female animals of control and 1000 mg/kg/bw/day dose group. All the changes observed in the control and 1000 mg/kg/bw/day dose treatment group animals were similar and the effect observed are not due to the test chemical.

Study 3: No biologically noteworthy changes in the incidences of neoplasms or non-neoplastic lesions occurred in any organ in male or female rats, except for a significant decrease in the incidence of uterine stromal polyps in the 6000 ppm females (control, 15/60; 3000 ppm, 18/60; 6000 ppm, 4/60; and 12000 ppm, 11/60),which was not considered chemical related.

Study 4: Visceral examination of the rats of control and other treated groups did not reveal any pathological abnormality.
Microscopic examination of control group and rats treated at 308, 556 and 1000 mg/kg revealed varying degree of pathological changes in different organs. This includes Liver: focal to multifocal minimal lymphocytic infiltration (Male: G1:1/5, G4:2/5; Female: G1: 1/5; G4: 2/5); focal minimal necrosis (Male: G1:1/5; Female: G1: 1/5); Kidneys: focal minimal lymphocytic infiltration (Male: G1:2/5; Female: G4:1/5); focal mild mineralization (Female: G1:1/5); Lungs: multifocal minimal lymphocytic infiltration (Male:G1:1/5, G4:1/5; Female: G1: 2/5, G4: 3/5); focal minimal histiocyte infiltration (Female: G1: 1/5, G4: 1/5); Heart: focal minimal lymphocytic infiltration (Male: G1:1/5, G4:1/5); Aorta: focal minimal aneurysm (Male:G1:1/5, G4:1/5); Mandibular Lymph Node: focal moderate cystic dilation of cortex (Female: G4:1/5); Stomach: focal mild squamous epithelium hyperplasia (Female: G1: 1/5); Mesenteric lymph node: focal moderate cystic dilation of cortex (Female:G1:1/5); Spleen: focal to diffuse minimal to mild extramedullary hematopoesis (Female: G1: 2/5, G4: 3/5); Thymus: mild to moderate atrophy (Female: G1:3/5, G4:4/5); focal mild cystic epithelial dilation (Male: G4:1/5; Female: G1: 1/5, G4:1/5); Trachea: focal to multifocal minimal to moderate Neutrophilic/lymphocytic infiltration (Male: G1:3/5, G4:3/5; Female: G1: 2/5, G4:1/5); Adrenals: unilateral accessory adrenocortical tissue (Male: G1:1/5, G4:1/5); Testes: focal to multifocal minimal to mild retention of mature sperm (Male: G1:4/13, G2:8/13, G3:8/13, G4:8/13); focal minimal to mild degeneration of seminiferous tubules (Male: G1:2/13, G2:1/13, G3:1/13, G4:1/13); focal to multifocal minimal sloughing of Pachytene Spermatocyte (Male: G1:2/13, G2:2/13, G3:2/13, G4:2/13); focal minimal sloughing of round spermatid (Male: G1:1/13, G2:1/13, G3:1/13, G4:1/13); focal mild infiltration of multinucleated giant cells (Male: G1:1/13); Seminal Vesicles: multifocal mild neutrophilic/lymphocytic infiltration (Male: G1:1/13); Prostate: focal moderate necrotic debris in lumen (Male: G2:1/13); Uterus: multifocal to diffuse mild reduction of stromal cells (Female: G1:1/13; G4:2/13); focal moderate necrosis (Female: G3:1/13); multifocal mild to moderate nodular hyperplasia (Female: G1:1/13; G2:1/13; G4:1/13); Cervix: focal minimal lymphocytic infiltration (Female: G2:1/13).
Histopathological findings: neoplastic:
not specified
Other effects:
no effects observed
Description (incidence and severity):
Study 4: No effect were observed on Number and sex of pups, stillbirths, live births, runts (pups that are significantly smaller than corresponding control pups), body weight and gross abnormalities of treated pups as compared to control.
Details on results:
No Data Available
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
clinical biochemistry
clinical signs
food consumption and compound intake
gross pathology
haematology
histopathology: non-neoplastic
mortality
organ weights and organ / body weight ratios
Remarks on result:
not determinable due to absence of adverse toxic effects
Critical effects observed:
no
Conclusions:
Based on all the observations and results, it was concluded that the NOAEL for the test chemical was considered to be 1000 mg/kg bw/day when Sprague-Dawley rats exposed orally for 28 days.
Executive summary:

Repeated Dose Toxicity Study:

The available data for all the repeated dose toxicity studies are as follows:

Study 2:

A sub-acute study was conducted to evaluate the toxic effects of repeated administration of the test chemical in male and female Sprague-Dawley rats by gavage. The test chemical was administered to 6 animals/sex/species in Corn oil at doses of 0, 250, 500 and 1000 mg/kg/BW/day for 28 days. All rats in 250, 500 and 1000 mg/kg/bw/day dose group survived throughout the study. The test chemical did not have any effect on mortality. Blood samples for Clinical Biochemistry and Hematology were collected. No abnormalities occurred in these parameters that could be directly attributed to the test chemical. Although significant change in relative weights of liver, ovaries and lungs of female were observed in 1000 mg/kg/ BW/day dose groups. However, these effects were disregarded since, no test chemical related gross pathological or histological changes were seen and findings were not dependent on the test chemical and hence considered to be of no toxicological importance. Thus, the NOEAL for repeated dose toxicity study was considered to be 1000 mg/kg/bw/day in male and female Sprague-Dawley rats when exposed to the test chemical by oral route for 28 days.

Study 3:

The test chemical is used as a flavoring agent in foods, as a fragrance in soaps and perfumes, as a solvent for cellulose acetate and nitrate, and as a component of printing inks and varnish removers. The toxicity of the test chemical is studied using the dosed feed route of administration. Groups of 60 male and 60 female F344/N rats were fed diets containing 0,3000, 6000, or 12000 ppm of the test chemical (0, 150, 300, 600 mg/kg body weight for 2 years. The LOAEL value of the test chemical was determined to be 12000 ppm based on the effect on reduced body weight and feed consumption.

Study 4:

In a combined repeated and reproductive developmental toxicity study, wistar male and female rats were treated with the test chemical in the concentration of 0, 308, 556 and 1000 mg/kg bw orally by gavage in Corn oil for 63 days. No mortality or morbidity and apparent treatment related clinical signs were observed any of the groups of animals throughout the study period. Detailed clinical examinations like Home cage observation, Handling observation and Open field observation of all animals were observed to be normal during study period. Number of rear, urine pools, fecal bolus in animals of all the test groups of both the sexes was comparable and did not show any treatment related significant difference as compared to the respective control groups. Body weight, percent body weight changes and feed consumption in animals of all the test groups of both the sexes was comparable and did not show any treatment related significant difference as compared to the respective control groups. The sensory reactivity measurements, Foot splay, fore limb and hind limb grip strength parameters and Motor activity were comparable and no treatment related changes were revealed in any of the animals of all treated groups as compare to the repective control groups. Similarly,No treatment related changes were observed inhematological and clinical chemistry parameters of treated male and female rats as compared to control.No test item related changes in estrous cyclicity and precoital interval were observed. There was statistically significant decrease in G3 (556 mg/kg body weight) as compared to control G1 (0 mg/kg body weight). This is not dose dependent hence not considered as treatment related. There was no statistically significant difference between the control and treatment groups in the maternal and pups parameters, except markedly decreased pregnancy index / fertility index in G4 (1000 mg/kg body weight), which was considered to be treatment related.In addition, no significant change in organ weight,External and visceral examination of treated and recovery groups as compared to control.Focal to multifocal minimal lymphocytic infiltration of male and female and focal minimal necrosis in male in liver, focal minimal lymphocytic infiltration of male and female and focal mild mineralization in female in kidney, multifocal minimal lymphocytic infiltration in male and female andfocal minimal histiocyte infiltration in female lungs, focal minimal lymphocytic infiltration in heart of male, focal minimal aneurysm in aorta of male, focal moderate cystic dilationof cortex of Mandibular Lymph Node in female,focal mildsquamous epitheliumhyperplasia stomach of female, focal moderate cystic dilation of cortex in Mesenteric lymph node, focalto diffuse minimal to mild extramedullary hematopoesis in spleen and mild to moderate atrophy in Thymus of female, focal to multifocal minimal to moderate Neutrophilic/lymphocytic infiltration of Trachea of male and female, unilateral accessory adrenocortical tissue of Adrenals and focal to multifocal minimal to mildretention of mature sperm,focal minimal to milddegeneration of seminiferous tubules,focal to multifocal minimalsloughing of Pachytene Spermatocyte,focal minimalsloughing of round spermatid andfocal mildinfiltration of multinucleated giant cells of Testes,multifocal mildneutrophilic/lymphocytic infiltration of Seminal Vesicles and focal moderate necrotic debris in lumen of Prostate observed in male rats, multifocal to diffuse mild reduction of stromal cells, focal moderate necrosis, multifocal mild to moderate nodular hyperplasia of Uterus and focal minimal lymphocytic infiltration of Cervix in female rats were observed. Lesions observed in liver, kidneys, lungs, heart, aorta, stomach, lymph nodes, spleen, thymus, trachea, adrenal gland and reproductive organs of high dose treated group rats are well comparable with respective control group rats and exhibited no dose relationship Therefore, No Observed Adverse Effect Level (NOAEL) is considered to be 1000 mg / kg body weight when Wistar male and female rats were orally treated with the test chemical.

Endpoint:
short-term repeated dose toxicity: oral
Remarks:
Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test
Data waiving:
other justification
Justification for data waiving:
other:
Justification for type of information:
The study is ongoing and this information will be submitted later based on ECHA communication/decision number CCH-D-2114536388-40-01/F.
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 from a Klimisch 2 source and thus provides a robust study summary.

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
Endpoint conclusion:
no study available

Repeated dose toxicity: inhalation - local 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
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:
exposure considerations
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
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - local effects

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: dermal
Data waiving:
exposure considerations
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
Endpoint conclusion:
no study available

Additional information

Repeated Dose Toxicity Study:

 

The available data fortherepeated doseoraltoxicity studies arethefollows:

 

Study 2:

A repeated dose 28-day oral toxicity study was performed to assess the toxicity profile of the test chemical when administered to Sprague -Dawley rats for 28 consecutive days. The test substance was administered via gavage to 6 rats/sex at the dose level of 250, 500 and 1000 mg/kg. Control animals received corn oil as vehicle. Rats were observed for general appearance and clinical signs daily, detailed clinical observation including home cage, handling and open field observations and behavioural assessments were performed weekly. Grips strength, reactivity to sensory stimuli and motor activity were assessed for three consecutive times during the last week of treatment. After completion of 28-day administration period, all rats were sacrificed and examined for gross lesion. At termination day (day 29th), blood samples were collected from all rats for haematology and clinical biochemistry. Vital organs together with male and female reproductive organs were collected, weighted and preserved for histopathological examination. Tissue samples of control and high dose groups were used for histopathological analyses. No case of death was observed during the 28-day administration period. Animals from control and different dose groups exhibited normal body weight gain and feed consumption throughout the dosing period. Daily clinical observation as well as the detailed clinical observations did not reveal any sign of toxicity in dosed rats during the administration period. Functional battery including the assessment of sensory reactivity to various stimuli, grip strength and the motor activity performed during the 4thweek of treatment did not reveal any abnormalities from treated groups when compared to controls. In addition, no treatment related and/or dose dependent changes were noted in haematological and clinical biochemistry parameters up to 1000 mg/kg body weight. External and visceral examination of all treated and control rats did not reveal any abnormality of pathological significance. The absolute and relative organ weights of brain, liver, kidney adrenals, testes, prostate with seminal vesicle and coagulation gland, epididymis, ovaries, uterus, heart, spleen, lung and thymus of dosed rats of either sex did not show treatment-related effect when compared to respective control groups. The relative weight of liver, lung and ovaries of female rats increased significantly at 1000 mg/kg but no related gross pathological, histopathological or haematological changes were seen thus, they were considered incidental alterations with no toxicological importance. In conclusion, the repeated oral exposure of the test chemical exerted no toxicologically significant changes in the morphology and function of a tissue/organ or have produced serious changes to the biochemistry or haematology of the organism when it was orally administered to male and female SD rats for 28 consecutive days. Hence, the NOAEL for repeated dose toxicity was 1000 mg/kg/day based on the absence of adverse effects observed.

 

Study 3:

Groups of 60 male and 60 female F344/N rats were fed diets containing 0, 3000, 6000 and 12000 ppm (0, 150, 300, 600 mg/kg) test chemical for 2 years. The doses selected for the 2-year feed study in F344/N rats were based on lower survival, mean body weights, and feed consumption, and on increased incidences of histopathologic brain lesions in 50 000 ppm male and female rats in the 13-week study. In the present 2-year feed study, the survival of dosed rats was similar to that of the controls. The mean body weights of the 12000 ppm males and females were minimally (approximately 5 %) lower than those of the controls throughout most of the study. The feed consumption by 12000 ppm males was slightly lower than that by the controls. Dietary levels of 3000, 6000, and 12000 ppm test substance were estimated to result in average daily consumption levels of 130, 260, and 510 mg/kg body weight(males) and 145, 290, and 575 mg/kg (females). No biologically significant changes in haematology or clinical chemistry parameters were found that could be attributed to the test chemical administration. Histopathological examination did not reveal compound-related increase of incidence of neoplasms or nonneoplastic lesions occurred in male or female F344/N rats receiving the test chemical for as long as 2 years. In conclusion, under the experimental conditions of the present 2-year feed study, it can be concluded that the test chemical exerted no carcinogenic activity when male or female F344/N rats were fed diet containing 3000, 6000, or 12000 ppm of test compound. In addition, as only slight decrease (ca. 5 %) was observed in the mean body weight and feed consumption of 12000 ppm male and female rats, it can be concluded that the oral administration of the test chemical did not induce serious adverse/toxic effect and therefore the NOAEL for repeat dose toxicity was 12000 ppm (575 and 510 mg/kg/day for male and female F344/N rats, respectively).

 

Study 4:

Combined Repeated Dose Toxicity Study (OECD TG 422) was conducted on male and female Wistar rats to assess the toxic properties of the test chemical following repeated oral administration. Groups of 13 rats/sex were administered via gavage at the dose level of 308, 556 and 1000 mg/kg of test chemical. Groups of 5 rats/sex were administered either 0 or 1000 mg/kg test chemical daily till the first scheduled sacrifice and kept for 14 days without treatment to detect delayed occurrence, or persistence of, or recovery from toxic effects. Control rats received corn oil as vehicle. All animals of both sexes were dosed 2 weeks prior to mating and during the mating period. Males were further dosed till day 47thday. Female rats were dosed during pregnancy till 4 day of postpartum. All animals were observed for mortality and morbidity twice daily. Rats were observed for general appearance and clinical signs daily, while detailed clinical examination performed outside the home cage was performed once before the first treatment, then weekly. Sensory reactivity to stimuli, assessment of grip strength, hind limb foot splay and motor activity assessment were conducted for 5 males and 5 females of each groups during the last week of treatment and for recovery groups during the last recovery week. The body weight and feed consumption were weighted weekly for both males and females. Pregnant females were weighted on day 0, 7, 14 and 20, post-partum day 0-1, on day 4 post-partum and before sacrifice. Haematology and clinical biochemistry were performed on 5 males and 5 females of each group, prior to necropsy and organs weights measurements. On termination day, all animals were subjected to gross pathological examination, vital organs were weighted and preserved for histopathological analyses. Histopathology was performed on tissue samples of five males and five females randomly selected from control and high-dose groups. No mortality and morbidity were observed in both treatment and recovery groups during the entire study period. No apparent treatment-related clinical sign was observed in the treatment and recovery groups. Sensory reactivity and motor activity measurements were comparable, and no treatment related changes were revealed at any dose level when compared to control groups. No treatment-related and dose-dependent changes in mean body weight were noted up to 1000 mg/kg both in treatment and recovery groups. However, the mean body weight change (%) of males decreased significantly during the entire treatment period (Day 1-46) at 1000 mg/kg and during the first two weeks of treatment at 556 mg/kg. Males from the recovery group also demonstrated significant decrease in the mean body weight changes (%) during day 1-29. Females gained significantly less weight during gestation (days 1-20), but the mean body weight change did not alter in the recovery group. The feed consumption of all treatment and recovery groups of both sexes were comparable and did not show any significant differences as compared with respective controls. In addition, no treatment related and/or dose dependent changes were noted in haematological and clinical biochemistry parameters up to 1000 mg/kg body weight. External and visceral examination of all treated and control rats including recovery groups did not reveal any abnormality of pathological significance. The absolute and relative organ weights of brain, adrenals, ovaries with oviduct, testes, epididymis, heart, liver, kidney spleen, thymus of dosed rats of either sex did not differ significantly when compared to respective control groups. Histopathology of liver, kidney, heart, thymus, aorta, stomach, adrenals, aorta and trachea did not reveal treatment-related and toxicologically relevant lesions at any dose level. In summary, male and female Wistar rats gained significantly less weight when administered with the test chemical up to 1000 mg/kg/day for 47-54 days. However, the repeated oral administration of the test substance did not produce serious toxic effect that can impair the function or morphology of specific target organ, neither produced serious changes to the biochemistry or haematology of the organism and therefore the test chemical was not classified as specific target organ toxicants following repeated oral exposure. The NOAEL for repeated dose toxicity was 556 mg/kg/day based on the significantly decreased weight gain.

Repeated Dose (Dermal) Toxicity Study:

A short-term dermal toxicity study does not need to be conducted because exposure of humans via dermal route in production and/or use is unlikely based on the provided thorough and rigorous exposure assessment. Also, the acute dermal toxicity value for test chemical (as provided in section 7.2.3) is >2000 mg/kg body weight. Given the use of the chemical; repeated exposure by the dermal route is unlikely since the use of gloves is common practice in industries. Therefore, it is expected that test chemical shall not exhibit toxicity via dermal route following exposure for 28 days. In addition, there is no data available that suggests that test chemical shall exhibit repeated dose toxicity by the dermal route and therefore this end point was considered for waiver.

Repeated Dose (Inhalation) Toxicity:

A short-term inhalation 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. Also, the given test chemical has very low vapor pressure, < 0.1333 Pa at 20°C, so the potential for the generation of inhalable vapor is very low. The normal conditions of use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalation route will be highly unlikely and therefore the end point of repeated inhalation toxicity was considered for waiver.

Justification for classification or non-classification

Weighting of evidence from several in vivo experimental studies (reviewed above) led to the conclusion that the repeatedoralexposure totest chemicaldid not produce toxicologically serious alterations in the morphology or function of target organs at or below the recommended guidance valuesand, consequentlythe test chemical is not classified asa toxicantaccording to the criteria of the CLP regulations.