Registration Dossier

Administrative data

Description of key information

Repeated dose toxicity: Oral

The No Observed Adverse Effect Level (NOAEL) was considered to be 1250 mg/kg/day, when male and female Wistar rats were treated with the given test chemical during repeated dose toxicity study.

Repeated dose toxicity: Inhalation

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. Also, the given test chemical has very low vapor pressure 0.00351 Pa (2.63e-5 mmHg), so the potential for the generation of inhalable vapors 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 inhalatory route will be highly unlikely and therefore this end point for repeated inhalation toxicity was considered for waiver.

 

Repeated dose toxicity: Dermal

A short-term toxicity study does not need to be conducted because exposure of humans via dermal in production and/or use is not likely as 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. 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:
chronic 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:
read-across source
Reason / purpose:
read-across source
Qualifier:
according to
Guideline:
other: As mentioned below
Principles of method if other than guideline:
WoE report is based on 2 repeated dose toxicity studies i.e. WoE-2 and WoE-3.
GLP compliance:
not specified
Limit test:
no
Species:
rat
Strain:
other: 2. Wistar W.74 (SPF) 3. Wistar
Sex:
male/female
Details on test animals and environmental conditions:
Source: Animals were procured from a CPCSEA approved Vendor [National Institute of Biosciences, Pune
Health Status:Healthy young adult animals were used for the study. Females were nulliparous and non-pregnant.
Age: 12 - 13 weeks at the start of estrous cycle evaluation.
Acclimatisation: Animals were acclimatised to the test conditions for 7 days prior to test item administration.
Animal Identification: During acclimatisation period, all the animals were identified by temporary tail marking with indelible ink and cage cards. Following allocation to the study, each animal was uniquely identified by micro toe pad tattooing and colour coded cage cards labelled with study no., study type, test system, sex, dose, group, animal number, dosing start date, experimental start and completion date. Pups were identified with body marking with permanent non-toxic marker pen.HusbandryHousingBefore 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 and during gestation and lactation only for females. Pregnant females were caged individually.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. 4-17 and 817 (Krishana Corncob Industries, Aurangabad)) 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 19.60 to 24.70°C the relative humidity was kept between 41.80 to 66.60 %. 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. 040817, 041017, 040917 and 040617 from approved supplier (Nutrivet Life Sciences, Pune) was offered ad libitum. The copy of composition, 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:
oral: gavage
Vehicle:
other: 2. 5% aqueous tylose 3.
Details on oral exposure:
2. VEHICLE
- Justification for use and choice of vehicle (if other than water): 5% aqueous tylose
- Concentration in vehicle: 0, 100, 330 and 1000 mg/kg bw
- Lot/batch no. (if required): 414 512

MAXIMUM DOSE VOLUME APPLIED: 10 ml/kg body weight.
3. 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
- Concentration in vehicle: 0, 750, 1000 and 1250 mg/kg bw
- Amount of vehicle (if gavage): 2 mL/100g body weight
- Lot/batch no. (if required): A1708001 and A1703001
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Specificity, Linearity, Precision (%RSD), Accuracy (% Recovery) and Homogeneity were analyze by using HPLC-UV.
Duration of treatment / exposure:
2. 13 weeks
3. More than 63 days
Frequency of treatment:
2. 7 days/week
3. Daily
Remarks:
2. Dose levels: 0, 100, 330 and 1000 mg/kg bw
3. 0, 750, 1000 and 1250 mg/kg bw/day
No. of animals per sex per dose:
2. Test: 30 animals [15♂, 15♀]/ dose group)
Control: 30 animals [15♂, 15♀]
3. Total: 124 0 mg/kg bw: 13 male , 13 female
750 mg/kg bw: 13 male , 13 female
1000 mg/kg bw: 13 male , 13 female
1250 mg/kg bw: 13 male , 13 female
Recovery group:0 mg/kg bw: 5 male , 5 female
1250 mg/kg bw: 5 male , 5female
Control animals:
yes
Details on study design:
2. No data
3. - Dose selection rationale: - Rationale for animal assignment (if not random): Randomization was done based on recent body weight, before first dosing. The animals were allocated to the different test groups using the ‘Group Allocation’ function in the MS Excel Add-in “Daniel’s XL Toolbar” (http://xltoolbox.sourceforge.net/). Individual body weights were considered within ± 20% of the groups mean.
Details of the randomization was documented in the raw data. - Other:
Observations and examinations performed and frequency:
2. CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Observations were carried out daily on all animals.
- Cage side observations checked: Mortality was observed.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Daily

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

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: Not specified
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Not specified
- Food consumption were recorded weekly.

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: Not specified

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Time schedule for examinations: recorded weekly.

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

HAEMATOLOGY: Yes
- Time schedule for collection of blood: week 6 and at termination of the study
- Anaesthetic used for blood collection: Not specified
- Animals fasted: Not specified
- How many animals: 5 males and 5 females of each group.
- Parameters checked in table [No.?] were examined. No data available

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: week 6 and at termination of the study
- Animals fasted: Not specified
- How many animals: 5 males and 5 females of each group.
- Parameters checked in table [No.?] were examined. No data available

URINALYSIS: Yes
- Time schedule for collection of urine: week 6 and at termination of the study
- Metabolism cages used for collection of urine: Not specified
- Animals fasted: Not specified
- Parameters checked in table [No.?] were examined.No data available

NEUROBEHAVIOURAL EXAMINATION: Not specified
- Time schedule for examinations:Not specified
- Dose groups that were examined:Not specified
- Battery of functions tested: sensory activity / grip strength / motor activity / other:Not specified

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: No data available
3. CAGE SIDE OBSERVATIONS: Yes - Time schedule:twice daily (morning and evening)
- Cage side observations checked in table [No.?] were included. morbidity and mortality, throughout the acclimatization and study period.

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. BODY WEIGHT: Yes - Time schedule for examinations:Males and females were weighed during randomization, 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: Five males and five females, randomly selected from each group, just prior to necropsy.
- Anaesthetic used for blood collection: Not specified
- Animals fasted: Yes, Animals were fasted overnight 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:Five males and five females, randomly selected from each group, just prior to necropsy
.- Animals fasted: Yes, Animals were fasted overnight 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), Globulin (Glob), Alb/ Glb (A:G), Bile acids, Thyroid hormones (Total T4 and TSH), Testosterone and Estrogen were examined. URINALYSIS: Not specified
- Time schedule for collection of urine:
- Metabolism cages used for collection of urine: Not specified
- Animals fasted: Not specified- Parameters checked in table [No.?] were examined.Not specified
NEUROBEHAVIOURAL EXAMINATION: yes
- Time schedule for examinations:During the last week of treatment and that of recovery groups
- Dose groups that were examined: five males and five females from control and treatment groups
- 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 were assess.
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:number and sex of pups, stillbirths, live births, runts (pups that are significantly smaller than corresponding control pups), and the presence of gross abnormalities.and Organ weight were examined.
Sacrifice and pathology:
2. Anatomic pathology examinations comprised organ weights, necropsy, histopathology.
GROSS PATHOLOGY: Yes
Organ weights (thymus, thyroid, heart, lung, liver, spleen, kidneys, adrenals, testes,ovaries) were determined for all animals. All animals were also subjected to macroscopic inspection after necropsy.

HISTOPATHOLOGY: Yes
Histopathological examination was carried out in 5 male and 5 female animals of the control- and the 1000 mg/kg-group. The organs looked at were:heart, lung, liver, pancreas, spleen, kidneys, pituitary, thyroid, thymus, adrenals,testes/epididymes, prostate gland, seminal vesicles, ovaries, uterus, salivary gland (head),oesophagus, stomach, intestines (duodenum, jejunum, ileum, olon), lymph nodes, bladder, brain, eyes, aorta, trachea, skeletal muscles, bones, bone marrow).
3. GROSS PATHOLOGY: Yes 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. This was followed by opening of the carcasses and topographic examination of different organs. This included careful examination of the external surface of the body, all orifices, cranial, thoracic and visceral cavities and their contents. Simultaneously gross lesions examination was performed in accordance with the Standard Operating Procedure (SOP) of the Laboratory. Number of implatation sites in uterus and number of corpora lutea of all pregnant females were counted during necropsy examination.Similarly, necropsy of terminally sacrificed and found dead pups during study period were conducted and gross pathological observations were recorded.

HISTOPATHOLOGY: Yes Tissues were preserved in 10 % neutral buffered formalin (NBF) (except eyes, which was fixed using Modified Davidson’s fluid; testes and epididymis, which were fixed in Bouin’s fluid for approximately 24 hours and subsequently preserved in 10 % NBF) for subsequent histopathological examination. Organ examined. Adrenals, Pancreas, Aorta, Peyer's Patches, Bone (femur) with joint , Pituitary, Brain, (cerebrum,cerebellum,mid brain), Prostate and Seminal vesicle with coagulating glands as a whole, Cecum Rectum, Colon, Salivary glands Duodenum, Sciatic Nerve, Epididymide, Skeletal muscle, Eyes with optic nerve, Skin, Spinal Cord (cervical, mid-thoracic and lumbar), Heart, Spleen, Ileum, Sternum with marrow, Jejunum, Stomach, Kidneys, Testes, Liver, Thymus, Lungs, Thyroid with Parathyroids, Mammary glands, Trachea, Mesenteric and Mandibular lymph node, Urinary Bladder, Oesophagus, Uterus, Ovaries with oviduct, Cervix with Vagina and Gross lesion (if any), were examined.
Statistics:
2. No data
3. 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):
2. No clinical signs of toxicity were seen in any group.

3. no effects observed - No apparent treatment related clinical signs were observed in any of the animals throughout the treatment period. However, statistically significant increase was noted in urine pool in group G2 in male on week 2 as compared to the G1 control group. In recovery female, statistically significant decrease was noted in rearing in G4-R on pretreatment, and increase in fecal bolus on week 1 as compared to the control recovery group G1-R. Statistically significant increase in urine pool in male G2 group on week 2 as compared to the control group G1.Detailed clinical examinations like Home cage observation, Handling observation and Open field observation of all animals were observed to be normal during study period in Main study.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:
mortality observed, non-treatment-related
Description (incidence):
2. Three mortalities (1♂, week 5, control group; 1♀, week 13, 330 mg/kg-group; 1♀, week 6, 1000 mg/kg-group) occurred intercurrently but were not substance related.

3. no mortality observed - No mortality or morbidity was observed in any of the groups of animals throughout the study period however, one male was found dead on Day 42 in group G2 at 750 mg/kg body weight and one male was found dead on Day 21 in group G4 at 1250 mg/kg body weight after dosing due to gavaging error.
Body weight and weight changes:
not specified
Description (incidence and severity):
3. no effects observed - No treatment related changes were noted in body weight and body weight gain upto 1250 mg/kg body weight.However, statistically significant decrease was noted in percent change in body weight with respect to day 1 in G4-R group on day 22 in male as compared to the control recovery group G1-R. 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):
2. Food consumption was the same in test and control groups.

3. no effects observed - No treatment related changes were noted upto 1250 mg/kg body weight. However, 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):
no effects observed
Description (incidence and severity):
2. Water consumption was the same in test and control groups.

3. not specified
Ophthalmological findings:
not specified
Haematological findings:
no effects observed
Description (incidence and severity):
2. Haematology results (total and differential blood count) of test animals did not differ from controls throughout the experiment.

3. no effects observed - No treatment related changes were noted in hematological parameters upto 1250 mg/kg body weight. However, statistically significant decreased was noted in male in aPTT in Group G4 at 1250 mg/kg body weight. In recovery males, statistically significant increase were noted in RBC and platelets at 1250 mg/kg body weight and statistically significant decrease was noted in PT at 1250 mg/kg body weight. The observed variations in aPTT, PT, RBC, and platelets were considered to be of no toxicological significance, of a minimal in nature and occurred in the absence of clear dose related response
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
2. Clinico-chemical parameters of test animals did not differ from controls throughout the experiment except for protein content in serum in females of the 1000 mg/kg-group, which was increased as compared to controls.However, the level found was within the biological variability range on record for female rats of this age and was therefore not considered substance related.

3. no effects observed - No treatment related chages were noted in clinical chemistry parameres upto 1250 mg/kg body weight. However, statistically significant decrease were noted in Albumin (Male: G3 at 1000 mg/kg body weight), creatinine (Male: G3 and G4 at 1000 and 1250 mg/kg body weight respectively), phosphorus (Male: G2 at 750 mg/kg body weight). In recovery males, statistically significant increase was noted in sodium at 1250 mg/kg body weight and statistically significant decrease were noted in AST and phosphorus at 1250 mg/kg body weight. The observed variations in clinical chemistry parameters were considered to be of no toxicological significance, of a minimal in nature and occurred in the absence of clear dose related response No treatment related changes were noted in hormonal analysis (T4, TSH, Testosteron and Estrogen) in any of the groups
Urinalysis findings:
not specified
Behaviour (functional findings):
no effects observed
Description (incidence and severity):
3. 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. However, statistically significant increase was noted in male in grip strength of fore limb in G3 group as compared to the control group G1. 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 groupThe 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):
2. Differences in organ weights, if observable at all (e.g. spleen in ♀'s), were small and not dose dependent.

3. no effects observed - At the end of treatment and recovery period, absolute and relative weight of organs of treated group rats of either sex did not differ significantly when compared to respective control group
Gross pathological findings:
no effects observed
Description (incidence and severity):
2. Necropsy-findings in the animals which had died intercurrently, revealed no pathological changes attributable to the test substance. Neither did the animals which were sacrificed pursuant to protocol at the end of the study exhibit any such changes.

3. no effects observed - 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. Visceral examination of the rats of control and other treated groups did not reveal any pathological abnormality except mild splenic enlargement of one male rat in G3 male and mild reduction of testis in one G4 male rat. Terminally sacrificed pups of all treated groups did not reveal any lesion of pathological significance in any of the group when compared with control group.
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
2. Histopathology did not reveal any organ change or –damage in any one of the dose groups.

3. no effects observed - Liver: multifocal minimal to mild Neutrophil/lymphocyte infiltration (Male: G1:2/5, G4:1/5; Female: G1/5), focal to multifocal mild degeneration (Male: G1:1/5, Female: G4/5), Kidneys: focal to multifocal mild lymphocyte infiltration (Male: G1:2/5, G4:1/5; Female: G1/5, G4:1/5); Lungs: multifocal minimal to mild lymphocyte infiltration (Male: G1:1/5, G4:1/5; Female: G1/5, G4:1/5), Heart: focal to multifocal minimal lymphocyte infiltration (Male: G1:1/5; Female: G4:1/5), Spleen: multifocal mild Extramedullary haematopoiesis (Male: G1:1/5), Adrenals: multifocal mild cytoplasmic vacuolation (male: G1:1/5), accessory adrenocortical tissue (Unilateral: Male: G1:2/5, G4:2/5; Female: G1:1/5), Testes: Male: focal mild seminiferous tubule degeneration (Male: G1: 2/13, G2:2/13, G4: 2/13, G1-R: 1/5); focal minimal to mild retention of mature sperm (Male: G1: 1/13, G2:1/13, G3: 1/13, G4: 1/13); focal minimal to mild sloughing of round spermatid/pachytene spermatocyte (Male: G1: 1/13, G2:1/13). Lesions observed in liver, kidneys, lungs, heart, adrenals, spleen and reproductive organs of high dose treated group rats are well comparable with respective control group rats and exhibited no dose relationship. Further these observed lesions are common in occurrence in rodents during toxicological studies (Boorman et al., 1990, Greaves, 2007, Greaves and Faccini, 1992; Haschek et al., 2002). Hence, occurrence of these lesions could be considered as spontaneous or incidental in nature and not to be attributed because of administration of the Test Item
Histopathological findings: neoplastic:
not specified
Other effects:
no effects observed
Description (incidence and severity):
3. No effect on number and sex of pups, stillbirths, live births, runts (pups that are significantly smaller than corresponding control pups), and the presence of gross abnormalities were observed.
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
food consumption and compound intake
water consumption and compound intake
haematology
clinical biochemistry
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
Remarks on result:
other: No toxic effect were observed
Remarks:
2
Dose descriptor:
NOAEL
Effect level:
1 250 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
haematology
clinical biochemistry
behaviour (functional findings)
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
Remarks on result:
other: No effect observed
Remarks:
3
Critical effects observed:
not specified
Conclusions:
The No Observed Adverse Effect Level (NOAEL) was considered to be 1250 mg/kg/day, when male and female Wistar rats were treated with the given test chemical during repeated dose toxicity study.
Executive summary:

Data available from the various sources was reviewed to determine the toxic nature of the given test chemical. The studies are as mentioned below:

 

The sub-chronic toxicity study was conducted by using the given test chemical as per OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents) in male and female Wistar rats at the concentration of 0, 100, 330 and 1000 mg/kgbw/day for 13 weeks via oral gavage route. The given test chemical was dissolved in 5% aqueous tylose and administered as 10 ml/kg bw via oral gavage route. Control animals received vehicle only. Animals were treated as given above and examined with respect to in-life-observations, clinical pathology and anatomic pathology. In-life-observations were carried out daily on all animals for mortalities and clinical signs of toxicity. Body weight and food-/water consumption were recorded weekly. Clinical pathology, viz. hematology, clinical chemistry and urinalysis, was performed in week 6 and at termination of the study using 5 males and 5 females of each group. Anatomic pathology examinations comprised organ weights, necropsy, and histopathology. Organ weights (thymus, thyroid, heart, lung, liver, spleen, kidneys, adrenals, testes, ovaries) were determined for all animals. All animals were also subjected to macroscopic inspection after necropsy. Histopathological examination was carried out in 5 male and 5 female animals of the control- and the 1000 mg/kg-group. The organs looked at were: heart, lung, liver, pancreas, spleen, kidneys, pituitary, thyroid, thymus, adrenals, testes/epididymes, prostate gland, seminal vesicles, ovaries, uterus, salivary gland (head), oesophagus, stomach, intestines (duodenum, jejunum, ileum, olon), lymph nodes, bladder, brain, eyes, aorta, trachea, skeletal muscles, bones, bone marrow). Three mortalities (1♂, week 5, control group; 1♀, week 13, 330 mg/kg-group; 1♀, week 6, 1000 mg/kg-group) occurred intercurrently but were not substance related. No clinical signs of toxicity were seen in any group. Food- and water consumption were the same in test- and control groups. Hematology results (total and differential blood count) of test animals did not differ from controls throughout the experiment. The same holds for clinico-chemical parameters, except for protein content in serum in females of the 1000 mg/kg-group, which was increased as compared to controls. However, the level found was within the biological variability range on record for female rats of this age and was therefore not considered substance related. Necropsy-findings in the animals which had died intercurrently, revealed no pathological changes attributable to the test substance. Neither did the animals which were sacrificed pursuant to protocol at the end of the study exhibit any such changes. Differences in organ weights, if observable at all (e.g. spleen in♀'s), were small and not dose dependent. Histopathology did not reveal any organ change or–damage in any one of the dose groups. Based on the above study, the toxicologically no adverse effect amount by repetitive oral administration of test substance for 13 weeks was observed and hence under this test condition NOAEL was considered to be1000 mg/kg bw/day.

 

In another Repeated Dose Oral Toxicity Study in combination with Reproduction / Developmental Toxicity study, Wistar male and female rat treated with test chemical in the concentration of 0, 750, 1000 and 1250 mg/kg bw orally by gavage for more than 63 days. No mortality or morbidity was observed in any of the groups of animals throughout the study period however, one male was found dead on Day 42 at 750 mg/kg body weight and one male was found dead on Day 21 at 1250 mg/kg body weight after dosing due to gavaging error. No apparent treatment related clinical signs were observed in any of the animals throughout the treatment period. However, statistically significant increase was noted in urine pool at 750 mg/kg bw in male on week 2 as compared to control group. In recovery female, statistically significant decrease was noted in rearing at 1250 mg/kg bw on pretreatment, and increase in fecal bolus on week 1 as compared to the control recovery. Detailed clinical examinations like Home cage observation, Handling observation and Open field observation of all animals were observed to be normal during study period in Main study. 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. No treatment related changes were noted in body weight and body weight gain upto 1250 mg/kg body weight. However, statistically significant decrease was noted in percent change in body weight with respect to day 1 at 1250 mg/kg bw on day 22 in male as compared to the control recovery. 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. No treatment related changes were noted upto 1250 mg/kg body weight. However, 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. Similarly, No treatment related changes were noted in hematological parameters upto 1250 mg/kg body weight. However, statistically significant decreased was noted in male in aPTT at 1250 mg/kg body weight. In recovery males, statistically significant increase were noted in RBC and platelets at 1250 mg/kg body weight and statistically significant decrease was noted in PT at 1250 mg/kg body weight. The observed variations in aPTT, PT, RBC, and platelets were considered to be of no toxicological significance, of a minimal in nature and occurred in the absence of clear dose related response. No treatment related charges were noted in clinical chemistry parameters upto 1250 mg/kg body weight. However, statistically significant decrease was noted in Albumin in male at 1000 mg/kg body weight, creatinine in male at 1000 and 1250 mg/kg body weight respectively, phosphorus in male at 750 mg/kg body weight. In recovery males, statistically significant increase was noted in sodium at 1250 mg/kg body weight and statistically significant decrease were noted in AST and phosphorus at 1250 mg/kg body weight. The observed variations in clinical chemistry parameters were considered to be of no toxicological significance, of a minimal in nature and occurred in the absence of clear dose related response. No treatment related changes were noted in hormonal analysis (T4, TSH, Testosteron and Estrogen) in any of the groups. 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. However, statistically significant increase was noted in male in grip strength of fore limb at 1000 mg/kg bw as compared to the control. 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. The above changes observed were inconsistent, hence considered as incidental and not attributed to the effect of test item administration. In addition, at the end of treatment and recovery period, absolute and relative weight of organs of treated group rats of either sex did not differ significantly when compared to respective control group. 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. Visceral examination of the rats of control and other treated groups did not reveal any pathological abnormality except mild splenic enlargement of one male rat at 1000 mg/kg bw male and mild reduction of testis in one at 125 mg/kg bw male rat.  Multifocal minimal to mild Neutrophil/lymphocyte infiltration in male (G1:2/5, G4:1/5) and female (G1/5), focal to multifocal mild degeneration in male (G1:1/5) and female (G4/5) in liver, focal to multifocal mild lymphocyte infiltration in male (G1:2/5, G4:1/5) and female (G1/5, G4:1/5) in Kidneys, multifocal minimal to mild lymphocyte infiltration in male (G1:1/5, G4:1/5) and Female (G1/5, G4:1/5) in Lungs, focal to multifocal minimal lymphocyte infiltration in male (G1:1/5) and Female (G4:1/5) in Heart, multifocal mild Extramedullary haematopoiesis in male (G1:1/5) in Spleen, multifocal mild cytoplasmic vacuolation in male (G1:1/5), Unilateral accessory adrenocortical tissue in male (G1:2/5, G4:2/5) and Female (G1:1/5) in Adrenals, focal mild seminiferous tubule degeneration in male (G1: 2/13, G2:2/13, G4: 2/13, G1-R: 1/5) in Testes; focal minimal to mild retention of mature sperm in male (G1: 1/13, G2:1/13, G3: 1/13, G4: 1/13); focal minimal to mild sloughing of round spermatid/pachytene spermatocyte in Male ( G1: 1/13, G2:1/13).  In control group G1 and treatment group G2, G3 and G4 all the females showed regular cyclicity i.e. 3-5 days estrous cycle. In cohabitation or mating period, all females from control and treated groups showed evidence of copulation i.e. sperm positive vaginal smear. All females showed precoital interval less than 5 days, except 4 females at 75 mg/kg bw which showed precoital interval more than 5 days. Therefore, NOAEL was considered to be 1250 mg/kg bw for P and F1 generation Wistar male and female rat treated with test chemical orally by gavage for more than 63 days.

 

Thus based on the above studies, the No Observed Adverse Effect Level (NOAEL) is considered to be 1250 mg / kg body weight and hence is not likely to classify as per the criteria mentioned in CLP regulation.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 250 mg/kg bw/day
Study duration:
chronic
Species:
rat
Quality of whole database:
Data is Klimicsh 2 and from handbook or collection of data.

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records
Reference
Endpoint:
chronic toxicity: inhalation
Data waiving:
other justification
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
Quality of whole database:
Waiver

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Link to relevant study records
Reference
Endpoint:
chronic toxicity: dermal
Data waiving:
other justification
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
Quality of whole database:
Waiver

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Repeated dose toxicity: Oral

Data available from the various sources was reviewed to determine the toxic nature of the given test chemical. The studies are as mentioned below:

 

The sub-chronic toxicity study was conducted by using the given test chemical as per OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents) in male and female Wistar rats at the concentration of 0, 100, 330 and 1000 mg/kgbw/day for 13 weeks via oral gavage route. The given test chemical was dissolved in 5% aqueous tylose and administered as 10 ml/kg bw via oral gavage route. Control animals received vehicle only. Animals were treated as given above and examined with respect to in-life-observations, clinical pathology and anatomic pathology. In-life-observations were carried out daily on all animals for mortalities and clinical signs of toxicity. Body weight and food-/water consumption were recorded weekly. Clinical pathology, viz. hematology, clinical chemistry and urinalysis, was performed in week 6 and at termination of the study using 5 males and 5 females of each group. Anatomic pathology examinations comprised organ weights, necropsy, and histopathology. Organ weights (thymus, thyroid, heart, lung, liver, spleen, kidneys, adrenals, testes, ovaries) were determined for all animals. All animals were also subjected to macroscopic inspection after necropsy. Histopathological examination was carried out in 5 male and 5 female animals of the control- and the 1000 mg/kg-group. The organs looked at were: heart, lung, liver, pancreas, spleen, kidneys, pituitary, thyroid, thymus, adrenals, testes/epididymes, prostate gland, seminal vesicles, ovaries, uterus, salivary gland (head), oesophagus, stomach, intestines (duodenum, jejunum, ileum, olon), lymph nodes, bladder, brain, eyes, aorta, trachea, skeletal muscles, bones, bone marrow). Three mortalities (1♂, week 5, control group; 1♀, week 13, 330 mg/kg-group; 1♀, week 6, 1000 mg/kg-group) occurred intercurrently but were not substance related. No clinical signs of toxicity were seen in any group. Food- and water consumption were the same in test- and control groups. Hematology results (total and differential blood count) of test animals did not differ from controls throughout the experiment. The same holds for clinico-chemical parameters, except for protein content in serum in females of the 1000 mg/kg-group, which was increased as compared to controls. However, the level found was within the biological variability range on record for female rats of this age and was therefore not considered substance related. Necropsy-findings in the animals which had died intercurrently, revealed no pathological changes attributable to the test substance. Neither did the animals which were sacrificed pursuant to protocol at the end of the study exhibit any such changes. Differences in organ weights, if observable at all (e.g. spleen in♀'s), were small and not dose dependent. Histopathology did not reveal any organ change or–damage in any one of the dose groups. Based on the above study, the toxicologically no adverse effect amount by repetitive oral administration of test substance for 13 weeks was observed and hence under this test condition NOAEL was considered to be1000 mg/kg bw/day.

 

In another Repeated Dose Oral Toxicity Study in combination with Reproduction / Developmental Toxicity study, Wistar male and female rat treated with test chemical in the concentration of 0, 750, 1000 and 1250 mg/kg bw orally by gavage for more than 63 days. No mortality or morbidity was observed in any of the groups of animals throughout the study period however, one male was found dead on Day 42 at 750 mg/kg body weight and one male was found dead on Day 21 at 1250 mg/kg body weight after dosing due to gavaging error. No apparent treatment related clinical signs were observed in any of the animals throughout the treatment period. However, statistically significant increase was noted in urine pool at 750 mg/kg bw in male on week 2 as compared to control group. In recovery female, statistically significant decrease was noted in rearing at 1250 mg/kg bw on pretreatment, and increase in fecal bolus on week 1 as compared to the control recovery. Detailed clinical examinations like Home cage observation, Handling observation and Open field observation of all animals were observed to be normal during study period in Main study. 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. No treatment related changes were noted in body weight and body weight gain upto 1250 mg/kg body weight. However, statistically significant decrease was noted in percent change in body weight with respect to day 1 at 1250 mg/kg bw on day 22 in male as compared to the control recovery. 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. No treatment related changes were noted upto 1250 mg/kg body weight. However, 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. Similarly, No treatment related changes were noted in hematological parameters upto 1250 mg/kg body weight. However, statistically significant decreased was noted in male in aPTT at 1250 mg/kg body weight. In recovery males, statistically significant increase were noted in RBC and platelets at 1250 mg/kg body weight and statistically significant decrease was noted in PT at 1250 mg/kg body weight. The observed variations in aPTT, PT, RBC, and platelets were considered to be of no toxicological significance, of a minimal in nature and occurred in the absence of clear dose related response. No treatment related charges were noted in clinical chemistry parameters upto 1250 mg/kg body weight. However, statistically significant decrease was noted in Albumin in male at 1000 mg/kg body weight, creatinine in male at 1000 and 1250 mg/kg body weight respectively, phosphorus in male at 750 mg/kg body weight. In recovery males, statistically significant increase was noted in sodium at 1250 mg/kg body weight and statistically significant decrease were noted in AST and phosphorus at 1250 mg/kg body weight. The observed variations in clinical chemistry parameters were considered to be of no toxicological significance, of a minimal in nature and occurred in the absence of clear dose related response. No treatment related changes were noted in hormonal analysis (T4, TSH, Testosteron and Estrogen) in any of the groups. 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. However, statistically significant increase was noted in male in grip strength of fore limb at 1000 mg/kg bw as compared to the control. 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. The above changes observed were inconsistent, hence considered as incidental and not attributed to the effect of test item administration. In addition, at the end of treatment and recovery period, absolute and relative weight of organs of treated group rats of either sex did not differ significantly when compared to respective control group. 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. Visceral examination of the rats of control and other treated groups did not reveal any pathological abnormality except mild splenic enlargement of one male rat at 1000 mg/kg bw male and mild reduction of testis in one at 125 mg/kg bw male rat.  Multifocal minimal to mild Neutrophil/lymphocyte infiltration in male (G1:2/5, G4:1/5) and female (G1/5), focal to multifocal mild degeneration in male (G1:1/5) and female (G4/5) in liver, focal to multifocal mild lymphocyte infiltration in male (G1:2/5, G4:1/5) and female (G1/5, G4:1/5) in Kidneys, multifocal minimal to mild lymphocyte infiltration in male (G1:1/5, G4:1/5) and Female (G1/5, G4:1/5) in Lungs, focal to multifocal minimal lymphocyte infiltration in male (G1:1/5) and Female (G4:1/5) in Heart, multifocal mild Extramedullary haematopoiesis in male (G1:1/5) in Spleen, multifocal mild cytoplasmic vacuolation in male (G1:1/5), Unilateral accessory adrenocortical tissue in male (G1:2/5, G4:2/5) and Female (G1:1/5) in Adrenals, focal mild seminiferous tubule degeneration in male (G1: 2/13, G2:2/13, G4: 2/13, G1-R: 1/5) in Testes; focal minimal to mild retention of mature sperm in male (G1: 1/13, G2:1/13, G3: 1/13, G4: 1/13); focal minimal to mild sloughing of round spermatid/pachytene spermatocyte in Male ( G1: 1/13, G2:1/13).  In control group G1 and treatment group G2, G3 and G4 all the females showed regular cyclicity i.e. 3-5 days estrous cycle. In cohabitation or mating period, all females from control and treated groups showed evidence of copulation i.e. sperm positive vaginal smear. All females showed precoital interval less than 5 days, except 4 females at 75 mg/kg bw which showed precoital interval more than 5 days. Therefore, NOAEL was considered to be 1250 mg/kg bw for P and F1 generation Wistar male and female rat treated with test chemical orally by gavage for more than 63 days.

 

Thus based on the above studies, the No Observed Adverse Effect Level (NOAEL) is considered to be 1250 mg / kg body weight and hence is not likely to classify as per the criteria mentioned in CLP regulation.

 

Repeated dose toxicity: Inhalation

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. Also, the given test chemical has very low vapor pressure 0.00351 Pa (2.63e-5 mmHg), so the potential for the generation of inhalable vapors 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 inhalatory route will be highly unlikely and therefore this end point for repeated inhalation toxicity was considered for waiver.

 

Repeated dose toxicity: Dermal

A short-term toxicity study does not need to be conducted because exposure of humans via dermal in production and/or use is not likely as 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. 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.

 

Justification for classification or non-classification

Based on the data available and applying the weight of evidence approach, the given test chemical does not exhibit toxic nature upon repeated exposure by oral route of exposure. Hence, it is not likely to classify as toxic as per the criteria mentioned in CLP regulation. For repeated inhalation and repeated dermal toxicity wavier was added so, not possible to classify.