Registration Dossier
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EC number: 202-336-5 | CAS number: 94-47-3
Developmental toxicity / teratogenicity
Administrative data
- Endpoint:
- developmental toxicity
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Justification for type of information:
- Data is from study report.
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 016
- Report date:
- 2016
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- other: OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
- Principles of method if other than guideline:
- The objective of this study was to provide evaluations of general and reproduction/ developmental toxicity endpoints associated with administration of repeated doses of test material in Wistar rats.
- GLP compliance:
- yes
- Limit test:
- no
Test material
- Reference substance name:
- Methyl phenyl acetate
- IUPAC Name:
- Methyl phenyl acetate
- Reference substance name:
- Methyl phenylacetate
- EC Number:
- 202-940-9
- EC Name:
- Methyl phenylacetate
- Cas Number:
- 101-41-7
- Molecular formula:
- C9H10O2
- IUPAC Name:
- methyl phenylacetate
- Test material form:
- liquid
- Details on test material:
- - Name of test material (as cited in study report): Methyl phenylacetate
- Molecular formula : C9H10O2
- Molecular weight : 150.177 g/mol
- Substance type: Organic
- Physical state: Liquid
- Lot/batch No.:2012051201
- Storage condition of test material: Room temperature (20° - 30 °C)
- Other:
Handling and Disposal
Safety precautions : Aprons, masks, caps, gloves and goggles were used to ensure the health and safety of the personnel.
Disposal : The remaining unused test item was disposed as per internal SOPs of sa-FORD and the same was documented in the raw data.
Constituent 1
Constituent 2
Test animals
- Species:
- rat
- Strain:
- Wistar
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: In-House Bred at sa-FORD, Animal Facility (CPCSEA Registration No. 1256/bc/09/CPCSEA)
- Females (if applicable) nulliparous and non-pregnant: Yes
- Age at study initiation: 12 - 13 weeks at the start of Oestrous Cycle evaluation.
- Weight at study initiation: Male: Minimum: 240 g Maximum: 315 g
Female: Minimum: 210 g Maximum: 260 g
- Fasting period before study: No data
- Housing: 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.
- Diet (e.g. ad libitum): A conventional laboratory pelleted diet was offered ad libitum
- Water (e.g. ad libitum): Aqua guard filtered drinking water in bottles was offered ad libitum
- Acclimation period: 20 days
DETAILS OF FOOD AND WATER QUALITY: A conventional laboratory pelleted diet was offered. Aqua guard filtered drinking water in bottles was offered.
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18.30 to 22.70 °C
- Humidity (%): 43.90 to 67.60%
- Air changes (per hr): 12 times per hour
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark
IN-LIFE DATES: From: To: November 16, 2015 to March 26, 2016
Administration / exposure
- Route of administration:
- oral: gavage
- Vehicle:
- corn oil
- Details on exposure:
- 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.
DIET PREPARATION
- Rate of preparation of diet (frequency): No data
- Mixing appropriate amounts with (Type of food): No data
- Storage temperature of food: No data
VEHICLE
- Justification for use and choice of vehicle (if other than water): Corn oil. The test chemical was soluble in corn oil
- Concentration in vehicle:
- Amount of vehicle (if gavage): 0.5 ml/100g body weight
- Lot/batch no. (if required): MR301015, MR161215
- Purity: No data - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- The analytical method was validated with respect to the following parameters.
Specificity:
The specificity will be evaluated by analysing the solvent used, standard solution, and sample solution.
Linearity:
The linearity was carried out by preparing and analyzing the standard solutions of at least 6 concentrations (covering the target analyte concentration i.e. 5 ppm,10 ppm, 25 ppm, 50 ppm, 75 ppm and 100 ppm ). A plot was drawn between the concentration and the response. The correlation coefficient, slope and intercept was calculated.
Assay accuracy and precision:
Assay accuracy and precision was carried out by fortifying the standard in vehicle at two levels (covering the target analyte concentration i.e., 10 ppm & 100 ppm). Five preparations were carried out at each concentration level selected. Two controls along with the assay accuracy samples were analysed. The mean, SD, % RSD was calculated. Assay accuracy was reported as the mean % recovery whereas the precision was reported as % RSD.
Homogeneity:
The homogeneity of the dose formulation prepared was determined by sampling and analyzing the formulation at top, middle and bottom layers. Sampling was done in two replicates from each layer.
Stability:
The stability of the prepared dose formulation was determined 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. - Details on mating procedure:
- - M/F ratio per cage: One male and one female (1:1)
- Length of cohabitation: Female rats were housed with same male until pregnancy occurs or two weeks elapsed.
- Proof of pregnancy: Mating was confirmed by observation of sperm positive vaginal smear. The day of detection of sperm positive vaginal smear was considered as day "0" of gestation.
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility.
- Further matings after two unsuccessful attempts: Yes, Re-mating of unsuccessfully paired female was done with proven male of the same group.
- After successful mating each pregnant female was caged (how): No data
- Any other deviations from standard protocol: No data - Duration of treatment / exposure:
- Total days: 64
All animals of both sexes were dosed 2 weeks prior to mating. Dosing was continued in both sexes during the mating period. Males were further dosed till 47th day . Females were dosed during pregnancy and upto day 4 post partum. - Frequency of treatment:
- Daily
- Duration of test:
- 64 days
Doses / concentrationsopen allclose all
- Dose / conc.:
- 0 mg/kg bw/day
- Remarks:
- G1 (Control Group)
- Dose / conc.:
- 308 mg/kg bw/day
- Remarks:
- G2 (Low Dose Group)
- Dose / conc.:
- 556 mg/kg bw/day
- Remarks:
- G3 (Mid Dose Group)
- Dose / conc.:
- 1 000 mg/kg bw/day
- Remarks:
- G4 (High Dose Group)
- Dose / conc.:
- 0 mg/kg bw/day
- Remarks:
- G1R (Control Recovery Group)
- Dose / conc.:
- 1 000 mg/kg bw/day
- Remarks:
- G4R (Recovery High Dose Group)
- No. of animals per sex per dose:
- Total: 124 ( 104 Test animals + 20 recovery animals)
Test animals:
0 mg/Kg bw: 13 males and 13 females
308 mg/Kg bw: 13 males and 13 females
556 mg/Kg bw: 13 males and 13 females
1000 mg/Kg bw: 13 males and 13 females
Recovery animals:
0 mg/Kg bw: 5 males and 5 females
1000 mg/Kg bw: 5 males and 5 females - Control animals:
- yes, concurrent vehicle
- Details on study design:
- - Dose selection rationale: The dose levels were selected based on the information provided by Sponsor.
- 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 validated software or the ‘Group Allocation’ function in the MS Excel Add-in “Daniel’s XL Toolbar” (http://xltoolbox.sourceforge.net/). Individual body weights will be considered within ± 20% of the groups mean.
- Other: No data
Examinations
- Maternal examinations:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily throughout the acclimatization and study period
- Cage side observations checked in table [No.?] were included. Mortality and morbidity
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: General clinical observations of animals of all groups were made once a day. Detailed clinical examinations were carried out once before the first treatment (to allow for within-subject comparisons) and weekly thereafter.
Observations included, but not be limited to, changes in skin, fur, eyes, mucous membranes, occurrence of secretions and excretions and autonomic activity (e.g. lacrimation, piloerection, pupil size, and unusual respiratory pattern). Changes in gait, posture and response to handling as well as the presence of clonic or tonic movements, stereotypies (e.g. excessive grooming, repetitive circling) or bizarre behaviour (e.g. self-mutilation, walking backwards).
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, During pre-mating, pregnancy and lactation, feed consumption were measured at least weekly. Feed consumption was not measured during mating period.
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Not specified
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): Not specified
- Time schedule for examinations: Not specified
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: just prior to necropsy at the end of the treatment and recovery periods
- Anaesthetic used for blood collection: Yes, Isoflurane anaesthesia
- Animals fasted: Yes, Animals were fasted overnight (approximately 16-18 hr) prior to blood collection
- How many animals: 5 males and 5 females
- 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), Activated Partial Thromboplastin time (aPTT).
CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: just prior to necropsy at the end of the treatment and recovery periods
- Animals fasted: Yes, Animals were fasted overnight (approximately 16-18 hr) prior to blood collection
- How many animals: 5 males and 5 females
- 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, Bile acids
URINALYSIS: Not specified
- Time schedule for collection of urine: Not specified
- Metabolism cages used for collection of urine: Not specified
- Animals fasted: Not specified
- Parameters checked in table [No.?] were examined. Not specified
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:
Functional Battery Observations: Sensory reactivity to stimuli, assessment of grip strength, hind limb foot splay and motor activity assessment were conducted for five males and five females from control and treatment groups, during the last week of treatment and that of recovery groups, in the last week of recovery period.
Animals were subjected to examinations of various functional parameters which included; motor activity measurements using OPTO–VARIMEX 4, an automated animal activity measuring system; fore limb and hind limb grip strength, using grip strength meter; hind limb foot splay record and sensory reactivity measurements. - Ovaries and uterine content:
- The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: No data
- Number of early resorptions: No data
- Number of late resorptions: No data
- Other: - Fetal examinations:
- - External examinations: Yes: all per litter
- Soft tissue examinations: No data
- Skeletal examinations: Yes:
- Head examinations: No data - Statistics:
- Raw data was analysed using statistical software “Sigma Plot 11.0”. 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
- Indices:
- Pregnancy index/fertility index was determined
- Historical control data:
- No data
Results and discussion
Results: maternal animals
General toxicity (maternal animals)
- Clinical signs:
- effects observed, non-treatment-related
- Description (incidence and severity):
- 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. - Dermal irritation (if dermal study):
- not specified
- Mortality:
- no mortality observed
- Description (incidence):
- No mortality or morbidity was observed in any animal of the control and treatment groups throughout the study period.
- Body weight and weight changes:
- effects observed, non-treatment-related
- Description (incidence and severity):
- 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):
- effects observed, non-treatment-related
- Description (incidence and severity):
- 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:
- no effects observed
- 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):
- 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 - Clinical biochemistry findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- 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):
- 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 respective 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):
- 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:
- effects observed, non-treatment-related
- Description (incidence and severity):
- External Findings: 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.
Internal Findings: Visceral examination of the rats of control and other treated groups did not reveal any pathological abnormality. - Neuropathological findings:
- not specified
- Histopathological findings: non-neoplastic:
- effects observed, non-treatment-related
- Description (incidence and severity):
- 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).
Microscopic examination of thyroid of male and female pups of control group and treated group did not revealed any lesion of pathological significance.
From the patho-morphological results presented, it is concluded that, the treatment of Methyl Phenyl acetate at 308, 556 and 1000 mg/kg body weight in male and female rats did not affect adversely and no alteration of pathological significance was observed in any of the organs including reproductive organs. - Histopathological findings: neoplastic:
- effects observed, non-treatment-related
- Description (incidence and severity):
- 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. Further these observed lesions are common in occurrence in rodents during toxicological studies. Hence, occurrence of these lesions could be considered as spontaneous or incidental in nature and not to be attributed to the administration of the Test Item.
- Other effects:
- not specified
Maternal developmental toxicity
- Number of abortions:
- not specified
- Pre- and post-implantation loss:
- not specified
- Total litter losses by resorption:
- not specified
- Early or late resorptions:
- not specified
- Dead fetuses:
- not specified
- Changes in pregnancy duration:
- not specified
- Description (incidence and severity):
- Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): not specified - Changes in number of pregnant:
- not specified
- Other effects:
- effects observed, treatment-related
- Description (incidence and severity):
- Pregnancy index was found to be 92.31, 84.62, 84.62 and 61.54 in G1, G2, G3 and G4 respectively. Marked decrease in Pregnancy index / Fertility index in G4(1000 mg/kg body weight) was considered to be treatment related.
Effect levels (maternal animals)
- Dose descriptor:
- NOAEL
- Effect level:
- 556 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Basis for effect level:
- behaviour (functional findings)
- body weight and weight gain
- clinical biochemistry
- clinical signs
- food consumption and compound intake
- food efficiency
- gross pathology
- haematology
- histopathology: neoplastic
- histopathology: non-neoplastic
- mortality
- organ weights and organ / body weight ratios
- Remarks on result:
- other: decrease in pregancy index was observed in 1000mg/kg bw dose group
Maternal abnormalities
- Abnormalities:
- not specified
- Localisation:
- not specified
Results (fetuses)
- Fetal body weight changes:
- no effects observed
- Description (incidence and severity):
- There was no statistically significant difference between the control (G1) and treatment groups for pups weight at birth and PND4 and weight gain at PND4.
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): not specified - Reduction in number of live offspring:
- no effects observed
- Changes in sex ratio:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Pups sex ratio (Male/Female) was found to be 55/57, 33/40, 43/58, and 21/26 in G1, G2, G3 and G4 respectively.
- Changes in litter size and weights:
- not specified
- Changes in postnatal survival:
- not specified
- External malformations:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Pups died during course of study revealed various lesions among the control and treated groups viz.,
External examination
Emaciated carcass (Male: G1:2/55, G2:1/44, G3:5/35; Female: G1:3/56, G2:1/30, G3:6/54);
Cannibalism (Male: G1:3/55, G3:2/35; Female: G1:2/56, G3:3/54);
Tearing of Neck Muscle (Female: G3:1/54; G4:1/18) and
Internal examination:
Absence of milk in stomach (Male: G1: 6/55, G2: 6/44, G3: 12/35, G4: 3/16; Female: G1: 8/56, G3: 14/54, G4: 2/18);
Blood clot in thoracic cavity (Male: G1: 2/55, G2: 3/44, G3: 1/35; Female: G1: 1/56, G3: 1/54, G4: 1/18);
Reddish discoloration of brain (Male: G1: 1/55, G2: 1/44, G3: 1/35; Female: G1: 1/56, G3: 3/54, G4: 1/18);
Reddish discoloration of lungs (Male: G1: 5/55, G2: 5/44, G3: 7/35, G4: 1/16; Female: G2: 1/30, G3: 10/54, G4: 2/18);
Paleness of liver (Male: G1: 1/55, G2: 2/44, G3: 1/35; Female: G3: 4/54, G4: 2/18);
Congested intestine (Female: G1: 1/56, G3: 1/54);
Autolytic changes (Female: G2: 1/30, G3: 2/54, G4: 1/18) - Skeletal malformations:
- not specified
- Visceral malformations:
- not specified
- Other effects:
- not specified
Effect levels (fetuses)
- Dose descriptor:
- NOAEL
- Effect level:
- 556 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- reduction in number of live offspring
- changes in sex ratio
- fetal/pup body weight changes
- external malformations
- Remarks on result:
- other: No developmental toxic effects was observed
Fetal abnormalities
- Abnormalities:
- not specified
- Localisation:
- other: not specified
Overall developmental toxicity
- Developmental effects observed:
- not specified
- Treatment related:
- not specified
- Relation to maternal toxicity:
- not specified
- Dose response relationship:
- not specified
- Relevant for humans:
- not specified
Any other information on results incl. tables
Table 1 Mortality and Morbidity
Sex: Male
Group | Treatment | Dose (mg/kg b.wt.) | No. of Animals/ Group | Observation During Study Period |
G1 | Control | 0 | 13 | NMM |
G2 | Low | 308 | 13 | NMM |
G3 | Mid | 556 | 13 | NMM |
G4 | High | 1000 | 13 | NMM |
G1-R | Control- Recovery | 0 | 5 | NMM |
G4-R | High- Recovery | 1000 | 5 | NMM |
Sex: Female
Group | Treatment | Dose (mg/kg b.wt.) | No. of Animals/ Group | Observation During Study Period |
G1 | Control | 0 | 13 | NMM |
G2 | Low | 308 | 13 | NMM |
G3 | Mid | 556 | 13 | NMM |
G4 | High | 1000 | 13 | NMM |
G1-R | Control -Recovery | 0 | 5 | NMM |
G4-R | High- Recovery | 1000 | 5 | NMM |
Keys:NMM = No mortality and morbidity observed, No.= Num
Table 2 Clinical Signs and Symptoms
Sex: Male
Group | Treatment | Dose (mg/kg b.wt.) | No. of Animals/ Group | Clinical Sign | Incidences During Study period |
G1 | Control | 0 | 13 | Normal | 13/13 |
G2 | Low | 308 | 13 | Normal | 13/13 |
G3 | Mid | 556 | 13 | Normal | 13/13 |
G4 | High | 1000 | 13 | Normal | 13/13 |
G1-R | Control -Recovery | 0 | 5 | Normal | 5/5 |
G4-R | High- Recovery | 1000 | 5 | Normal | 5/5 |
Sex: Female
Group | Treatment | Dose (mg/kg b.wt.) | No. of Animals/ Group |
Clinical Sign | Incidences During Study period |
G1 | Control | 0 | 13 | Normal | 13/13 |
G2 | Low | 308 | 13 | Normal | 13/13 |
G3 | Mid | 556 | 13 | Normal | 13/13 |
G4 | High | 1000 | 13 | Normal | 13/13 |
G1-R | Control -Recovery | 0 | 5 | Normal | 5/5 |
G4-R | High- Recovery | 1000 | 5 | Normal | 5/5 |
Key:No.= Number
Table 3 Summary of Detailed Clinical Examinations
Week:Pre-Treatment Sex:Male
Group (N) | G1 (13) | G2 (13) | G3 (13) | G4 (13) | G1-R (5) | G4-R (5) | |
Dose (mg/kg body weight) | 0 | 308 | 556 | 1000 | 0 | 1000 | |
Parameter | Observation | No. of Animals Showing Observation | |||||
Posture | Curled up, often asleep | 1 | 3 | 2 | 3 | 1 | 1 |
Sitting B | 0 | 2 | 2 | 2 | 0 | 0 | |
Sitting C | 1 | 2 | 1 | 1 | 2 | 1 | |
Sitting A | 6 | 2 | 2 | 3 | 1 | 2 | |
Rearing | 5 | 4 | 6 | 4 | 1 | 1 | |
Convulsions | Absent | 13 | 13 | 13 | 13 | 5 | 5 |
Ease of removing from the cage | Very easy | 11 | 11 | 12 | 10 | 4 | 5 |
Easy | 2 | 2 | 1 | 3 | 1 | 0 | |
Moderately difficult | 0 | 0 | 0 | 0 | 0 | 0 | |
Handling reactivity | Easy | 13 | 13 | 13 | 13 | 5 | 5 |
Moderately easy | 0 | 0 | 0 | 0 | 0 | 0 | |
Palpebral closure | Eyelids wide open | 13 | 13 | 13 | 13 | 5 | 5 |
Lacrimation | None (No lacrimation) | 13 | 13 | 13 | 13 | 5 | 5 |
Eye Examination | Absent | 13 | 13 | 13 | 13 | 5 | 5 |
Piloerection | Absent | 13 | 13 | 13 | 13 | 5 | 5 |
Skin Examination | Absent | 13 | 13 | 13 | 13 | 5 | 5 |
Salivation | None | 13 | 13 | 13 | 13 | 5 | 5 |
Gait | Normal | 13 | 13 | 13 | 13 | 5 | 5 |
Mobility | Normal | 13 | 13 | 13 | 13 | 5 | 5 |
Arousal | Normal | 13 | 13 | 13 | 13 | 5 | 5 |
Respiration | Normal | 13 | 13 | 13 | 13 | 5 | 5 |
Tonic Movement | Absent | 13 | 13 | 13 | 13 | 5 | 5 |
Clonic Movement | Absent | 13 | 13 | 13 | 13 | 5 | 5 |
Stereotypy | Absent | 13 | 13 | 13 | 13 | 5 | 5 |
Bizzare Behaviour | Absent | 13 | 13 | 13 | 13 | 5 | 5 |
Number of Rears | Mean | 11.5 | 10.2 | 7.5↓ | 7.2↓ | 7.2 | 6.4 |
Vocalization Count | Mean | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 |
No. of urine pools | Mean | 1.2 | 0.9 | 3.5↑ | 4.2↑ | 3.6 | 3.8 |
No. of faecal bolus | Mean | 1.2 | 0.5 | 3.8↑ | 1.8 | 2.4 | 1.4 |
Keys:N = Number of animals in group, No. = Number,↓ = Statistically Significant Decrease (atp < 0.05),↑=Statistically Significant Increase (atp < 0.05)
Table 3 Summary of Detailed Clinical Examinations Continued
Week:Pre-Treatment Sex:Female
Group (N) | G1 (13) | G2 (13) | G3 (13) | G4 (13) | G1-R (5) | G4-R (5) | |
Dose (mg/kg body weight) | 0 | 308 | 556 | 1000 | 0 | 1000 | |
Parameter | Observation | No. of Animals Showing Observation | |||||
Posture | Curled up, often asleep | 3 | 4 | 2 | 2 | 1 | 1 |
Sitting B | 2 | 1 | 4 | 2 | 1 | 1 | |
Sitting C | 2 | 0 | 1 | 1 | 1 | 0 | |
Sitting A | 2 | 2 | 3 | 1 | 1 | 1 | |
Rearing | 4 | 6 | 3 | 7 | 1 | 2 | |
Convulsions | Absent | 13 | 13 | 13 | 13 | 5 | 5 |
Ease of removing from the cage | Very easy | 10 | 12 | 12 | 11 | 4 | 4 |
Easy | 3 | 1 | 1 | 2 | 1 | 1 | |
Moderately difficult | 0 | 0 | 0 | 0 | 0 | 0 | |
Handling reactivity | Easy | 13 | 13 | 13 | 13 | 5 | 5 |
Moderately easy | 0 | 0 | 0 | 0 | 0 | 0 | |
Palpebral closure | Eyelids wide open | 13 | 13 | 13 | 13 | 5 | 5 |
Lacrimation | None (No lacrimation) | 13 | 13 | 13 | 13 | 5 | 5 |
Eye Examination | Absent | 13 | 13 | 13 | 13 | 5 | 5 |
Piloerection | Absent | 13 | 13 | 13 | 13 | 5 | 5 |
Skin Examination | Absent | 13 | 13 | 13 | 13 | 5 | 5 |
Salivation | None | 13 | 13 | 13 | 13 | 5 | 5 |
Gait | Normal | 13 | 13 | 13 | 13 | 5 | 5 |
Mobility | Normal | 13 | 13 | 13 | 13 | 5 | 5 |
Arousal | Normal | 13 | 13 | 13 | 13 | 5 | 5 |
Respiration | Normal | 13 | 13 | 13 | 13 | 5 | 5 |
Tonic Movement | Absent | 13 | 13 | 13 | 13 | 5 | 5 |
Clonic Movement | Absent | 13 | 13 | 13 | 13 | 5 | 5 |
Stereotypy | Absent | 13 | 13 | 13 | 13 | 5 | 5 |
Bizzare Behaviour | Absent | 13 | 13 | 13 | 13 | 5 | 5 |
Number of Rears | Mean | 6.2 | 10.0↑ | 11.2↑ | 10.5↑ | 10.4 | 11.2 |
Vocalization Count | Mean | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 |
No. of urine pools | Mean | 1.0 | 1.3 | 1.2 | 1.1 | 1.2 | 1.8 |
No. of faecal bolus | Mean | 0.9 | 0.8 | 1.2 | 1.2 | 1.2 | 1.0 |
Keys:N = Number of animals in group, No.= Number,↑= Statistically Significant Increase (atp < 0.05)
Table 3 Summary of Detailed Clinical Examinations Continued
Week:1st Sex:Male
Group (N) | G1 (13) | G2 (13) | G3 (13) | G4 (13) | G1-R (5) | G4-R (5) | |
Dose (mg/kg body weight) | 0 | 308 | 556 | 1000 | 0 | 1000 | |
Parameter | Observation | No. of Animals Showing Observation | |||||
Posture | Curled up, often asleep | 3 | 3 | 6 | 5 | 2 | 2 |
Sitting B | 8 | 8 | 6 | 7 | 3 | 2 | |
Sitting C | 1 | 2 | 1 | 1 | 0 | 1 | |
Sitting A | 1 | 0 | 0 | 0 | 0 | 0 | |
Rearing | 0 | 0 | 0 | 0 | 0 | 0 | |
Convulsions | Absent | 13 | 13 | 13 | 13 | 5 | 5 |
Ease of removing from the cage | Very easy | 11 | 12 | 13 | 10 | 5 | 5 |
Easy | 2 | 0 | 0 | 3 | 0 | 0 | |
Moderately difficult | 0 | 1 | 0 | 0 | 0 | 0 | |
Handling reactivity | Easy | 13 | 12 | 12 | 13 | 5 | 5 |
Moderately easy | 0 | 1 | 1 | 0 | 0 | 0 | |
Palpebral closure | Eyelids wide open | 13 | 13 | 13 | 13 | 5 | 5 |
Lacrimation | None (No lacrimation) | 13 | 13 | 13 | 13 | 5 | 5 |
Eye Examination | Absent | 13 | 13 | 13 | 13 | 5 | 5 |
Piloerection | Absent | 13 | 13 | 13 | 13 | 5 | 5 |
Skin Examination | Absent | 13 | 13 | 13 | 13 | 5 | 5 |
Salivation | None | 13 | 13 | 13 | 13 | 5 | 5 |
Gait | Normal | 13 | 13 | 13 | 13 | 5 | 5 |
Mobility | Normal | 13 | 13 | 13 | 13 | 5 | 5 |
Arousal | Normal | 13 | 13 | 13 | 13 | 5 | 5 |
Respiration | Normal | 13 | 13 | 13 | 13 | 5 | 5 |
Tonic Movement | Absent | 13 | 13 | 13 | 13 | 5 | 5 |
Clonic Movement | Absent | 13 | 13 | 13 | 13 | 5 | 5 |
Stereotypy | Absent | 13 | 13 | 13 | 13 | 5 | 5 |
Bizzare Behaviour | Absent | 13 | 13 | 13 | 13 | 5 | 5 |
Number of Rears | Mean | 5.7 | 6.3 | 5.5 | 7.3 | 6.0 | 6.2 |
Vocalization Count | Mean | 0.0 | 0.1 | 0.0 | 0.0 | 0.0 | 0.0 |
No. of urine pools | Mean | 2.9 | 2.8 | 3.2 | 2.7 | 3.0 | 4.6 |
No. of faecal bolus | Mean | 1.5 | 1.4 | 2.4 | 1.8 | 1.4 | 1.6 |
Keys:N = Number of animals in group, No.= Number
Table 3 Summary of Detailed Clinical Examinations Continued
Week:1st Sex:Female
Group (N) | G1 (13) | G2 (13) | G3 (13) | G4 (13) | G1-R (5) | G4-R (5) | |
Dose (mg/kg body weight) | 0 | 308 | 556 | 1000 | 0 | 1000 | |
Parameter | Observation | No. of Animals Showing Observation | |||||
Posture | Curled up, often asleep | 0 | 5 | 8 | 6 | 0 | 1 |
Sitting B | 5 | 5 | 4 | 5 | 3 | 2 | |
Sitting C | 4 | 2 | 1 | 1 | 2 | 2 | |
Sitting A | 4 | 1 | 0 | 1 | 0 | 0 | |
Rearing | 0 | 0 | 0 | 0 | 0 | 0 | |
Convulsions | Absent | 13 | 13 | 13 | 13 | 5 | 5 |
Ease of removing from the cage | Very easy | 11 | 13 | 11 | 13 | 5 | 4 |
Easy | 2 | 0 | 2 | 0 | 0 | 1 | |
Moderately difficult | 0 | 0 | 0 | 0 | 0 | 0 | |
Handling reactivity | Easy | 11 | 13 | 10 | 13 | 5 | 4 |
Moderately easy | 2 | 0 | 3 | 0 | 0 | 1 | |
Palpebral closure | Eyelids wide open | 13 | 13 | 13 | 13 | 5 | 5 |
Lacrimation | None (No lacrimation) | 13 | 13 | 13 | 13 | 5 | 5 |
Eye Examination | Absent | 13 | 13 | 13 | 13 | 5 | 5 |
Piloerection | Absent | 13 | 13 | 13 | 13 | 5 | 5 |
Skin Examination | Absent | 13 | 13 | 13 | 13 | 5 | 5 |
Salivation | None | 13 | 13 | 13 | 13 | 5 | 5 |
Gait | Normal | 13 | 13 | 13 | 13 | 5 | 5 |
Mobility | Normal | 13 | 13 | 13 | 13 | 5 | 5 |
Arousal | Normal | 13 | 13 | 13 | 13 | 5 | 5 |
Respiration | Normal | 13 | 13 | 13 | 13 | 5 | 5 |
Tonic Movement | Absent | 13 | 13 | 13 | 13 | 5 | 5 |
Clonic Movement | Absent | 13 | 13 | 13 | 13 | 5 | 5 |
Stereotypy | Absent | 13 | 13 | 13 | 13 | 5 | 5 |
Bizzare Behaviour | Absent | 13 | 13 | 13 | 13 | 5 | 5 |
Number of Rears | Mean | 10.5 | 9.9 | 10.2 | 10.4 | 10.2 | 10.2 |
Vocalization Count | Mean | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 |
No. of urine pools | Mean | 2.0 | 1.2 | 1.5 | 1.3 | 2.2 | 1.4 |
No. of faecal bolus | Mean | 1.1 | 1.2 | 1.1 | 1.4 | 1.6 | 1.2 |
Keys:N = Number of animals in group, No.= Number
Table 3 Summary of Detailed Clinical Examinations Continued
Week:7th
Sex | Male | Female | |||
Group (N) | G1-R (5) | G4-R (5) | G1-R (5) | G4-R (5) | |
Dose (mg/kg body weight) | 0 | 1000 | 0 | 1000 | |
Parameter | Observation | No. of Animals Showing Observation | |||
Posture | Curled up, often asleep | 2 | 3 | 2 | 3 |
Sitting B | 1 | 1 | 2 | 1 | |
Sitting C | 1 | 0 | 1 | 0 | |
Sitting A | 1 | 1 | 0 | 1 | |
Rearing | 0 | 0 | 0 | 0 | |
Convulsions | Absent | 5 | 5 | 5 | 5 |
Ease of removing from the cage | Very easy | 4 | 3 | 4 | 2 |
Easy | 1 | 2 | 1 | 3 | |
Moderately difficult | 0 | 0 | 0 | 0 | |
Handling reactivity | Easy | 5 | 4 | 3 | 3 |
Moderately easy | 0 | 1 | 2 | 2 | |
Palpebral closure | Eyelids wide open | 5 | 5 | 5 | 5 |
Lacrimation | None (No lacrimation) | 5 | 5 | 5 | 5 |
Eye Examination | Absent | 5 | 5 | 5 | 5 |
Piloerection | Absent | 5 | 5 | 5 | 5 |
Skin Examination | Absent | 5 | 5 | 5 | 5 |
Salivation | None | 5 | 5 | 5 | 5 |
Gait | Normal | 5 | 5 | 5 | 5 |
Mobility | Normal | 5 | 5 | 5 | 5 |
Arousal | Normal | 5 | 5 | 5 | 5 |
Respiration | Normal | 5 | 5 | 5 | 5 |
Tonic Movement | Absent | 5 | 5 | 5 | 5 |
Clonic Movement | Absent | 5 | 5 | 5 | 5 |
Stereotypy | Absent | 5 | 5 | 5 | 5 |
Bizzare Behaviour | Absent | 5 | 5 | 5 | 5 |
Number of Rears | Mean | 4.2 | 5.6 | 9.6 | 11.2 |
Vocalization Count | Mean | 0.0 | 0.0 | 0.0 | 0.0 |
No. of urine pools | Mean | 2.0 | 0.6 | 2.0 | 3.2 |
No. of faecal bolus | Mean | 2.8 | 1.2 | 1.2 | 1.4 |
Keys:N = Number of animals in group, No.= Number
Table 4 Mean Body Weight (g)
Sex: Male
Group (N) | G1 (13) | G2 (13) | G3 (13) | G4 (13) | ||||
Dose (mg/kg b. wt.) | 0 | 308 | 556 | 1000 | ||||
Day | Mean | SD | Mean | SD | Mean | SD | Mean | SD |
Day 1 | 247.38 | 11.20 | 262.38 | 20.32 | 261.62 | 18.86 | 261.92 | 17.03 |
Day 8 | 282.31 | 9.33 | 294.92 | 19.40 | 289.31 | 20.12 | 278.69 | 19.98 |
Day 14 | 315.00 | 13.00 | 324.38 | 21.35 | 316.15 | 20.45 | 299.15 | 21.37 |
Day 21 | 333.15 | 15.35 | 339.46 | 25.50 | 336.23 | 22.65 | 316.23 | 18.79 |
Day 28 | 350.08 | 18.06 | 362.62 | 30.37 | 358.54 | 27.49 | 334.15 | 20.60 |
Day 30 | 355.77 | 18.46 | 368.00 | 30.44 | 364.54 | 27.22 | 331.62↓ | 19.88 |
Day 37 | 370.77 | 22.36 | 381.92 | 30.46 | 381.77 | 31.58 | 351.62 | 24.13 |
Day 44 | 393.31 | 26.08 | 407.69 | 34.24 | 402.23 | 34.23 | 368.23 | 26.45 |
Day 46 | 397.23 | 24.79 | 414.77 | 34.07 | 405.38 | 34.10 | 370.92 | 26.71 |
Day 47 (Fasting) | 372.00 | 25.57 | 391.08 | 33.83 | 383.38 | 33.84 | 350.15 | 26.15 |
Period: Pre-mating Sex: Female
Group (N) | G1 (13) | G2 (13) | G3 (13) | G4 (13) | ||||
Dose (mg/kg b. wt.) | 0 | 308 | 556 | 1000 | ||||
Day | Mean | SD | Mean | SD | Mean | SD | Mean | SD |
Day 1 | 226.62 | 6.50 | 231.54 | 7.34 | 230.15 | 6.73 | 228.77 | 7.90 |
Day 8 | 229.85 | 8.37 | 233.46 | 7.85 | 233.54 | 9.00 | 227.92 | 7.39 |
Day 14 | 232.77 | 8.32 | 236.92 | 8.23 | 237.00 | 9.65 | 232.62 | 8.01 |
Keys:N = Number of animals in group, g= gram, SD = Standard deviation,↓= Statistically Significant Decrease (atp<0.05).
Table 4 Mean Body Weight (g) Continued
Period: Gestation Sex: Female
Group (N) | G1 (12) | G2 (11) | G3 (11) | G4 (8) | ||||
Dose (mg/kg b. wt.) | 0 | 308 | 556 | 1000 | ||||
Day | Mean | SD | Mean | SD | Mean | SD | Mean | SD |
Day 0 | 232.83 | 10.14 | 236.36 | 9.15 | 237.64 | 8.02 | 231.13 | 6.98 |
Day 7 | 245.25 | 18.20 | 249.91 | 8.83 | 251.36 | 9.75 | 237.13 | 8.56 |
Day 14 | 268.58 | 26.15 | 268.55 | 7.10 | 275.36 | 15.50 | 248.50 | 9.89 |
Day 20 | 302.00 | 37.56 | 302.45 | 13.13 | 310.64 | 27.21 | 264.50↓ | 11.74 |
Period: Post-Partum Sex: Female
Group (N) | G1 (12) | G2 (11) | G3 (11) | G4 (8) | ||||
Dose (mg/kg b. wt.) | 0 | 308 | 556 | 1000 | ||||
Day | Mean | SD | Mean | SD | Mean | SD | Mean | SD |
Day 0/1 | 240.83 | 25.04 | 254.36 | 26.47 | 252.27 | 19.08 | 228.63 | 15.00 |
Day 4 | 242.58 | 24.39 | 251.27 | 26.88 | 256.27 | 22.95 | 225.50 | 15.12 |
Day 5 (Fasting) | 226.83 | 20.94 | 236.18 | 24.64 | 240.00 | 20.97 | 214.25 | 13.44 |
Keys:N= Number of animals in group, g= gram, SD = Standard deviation,↓= Statistically Significant Decrease (atp<0.05)
Table 6 Mean Feed Consumption (g/day/animal)
Sex: Male
Group (N) | G1 (13) | G2 (13) | G3 (13) | G4 (13) | ||||
Dose (mg/kg b. wt.) | 0 | 308 | 556 | 1000 | ||||
Day | Mean | SD | Mean | SD | Mean | SD | Mean | SD |
Day 1-8 | 23.01 | 1.29 | 22.97 | 1.66 | 22.33 | 1.67 | 20.99 | 1.93 |
Day 8-14 | 24.39 | 1.69 | 23.48 | 0.90 | 22.81 | 1.55 | 22.52 | 1.10 |
Day 30-37 | 23.71 | 1.20 | 23.19 | 1.22 | 23.50 | 1.64 | 23.52 | 1.38 |
Day 37-44 | 29.33 | 3.03 | 28.84 | 4.00 | 28.07 | 4.45 | 26.89 | 4.34 |
Day 44-46 | 26.63 | 1.27 | 25.45 | 1.73 | 25.18 | 0.98 | 25.15 | 1.92 |
Period: Pre-mating Sex: Female
Group (N) | G1 (13) | G2 (13) | G3 (13) | G4 (13) | ||||
Dose (mg/kg b. wt.) | 0 | 308 | 556 | 1000 | ||||
Day | Mean | SD | Mean | SD | Mean | SD | Mean | SD |
Day 1-8 | 12.76 | 1.21 | 13.08 | 0.41 | 13.15 | 0.85 | 11.80 | 0.68 |
Day 8-14 | 11.23 | 1.04 | 11.10 | 0.67 | 11.80 | 0.48 | 10.71 | 0.96 |
Keys:N = Number of animals in group, g= gram, SD = Standard deviation,
Table 6 Mean Feed Consumption (g/day/animal) Continued
Period: Gestation Sex: Female
Group (N) | G1 (12) | G2 (11) | G3 (11) | G4 (8) | ||||
Dose (mg/kg b. wt.) | 0 | 308 | 556 | 1000 | ||||
Day | Mean | SD | Mean | SD | Mean | SD | Mean | SD |
Day 0-7 | 15.93 | 3.64 | 16.18 | 2.29 | 16.16 | 3.06 | 13.55 | 1.59 |
Day 7-14 | 17.96 | 3.53 | 18.26 | 1.76 | 18.05 | 2.50 | 15.29 | 1.97 |
Day 14-20 | 19.46 | 4.85 | 17.44 | 1.98 | 17.23 | 3.35 | 13.71↓ | 1.35 |
Period: Gestation and Post-Partum Sex: Female
Group (N) | G1 (12) | G2 (11) | G3 (11) | G4 (8) | ||||
Dose (mg/kg b. wt.) | 0 | 308 | 556 | 1000 | ||||
Day | Mean | SD | Mean | SD | Mean | SD | Mean | SD |
Day 20 -Day 5 PP | 18.40 | 5.22 | 18.82 | 4.69 | 18.25 | 5.56 | 13.12 | 2.06 |
Keys:g= gram, SD = Standard deviation, N = Number of animals in group, PP= Post Partum↓= Statistically Significant Decrease (atp<0.05)
Applicant's summary and conclusion
- Conclusions:
- No Observed Adverse Effect Level (NOAEL) is considered to be 556 mg/kg bw. When male and female wistar rats were treated with test material orally.
- Executive summary:
Combined repeated dose and reproductive-developmental toxicity study was to provide evaluations of general and reproduction/ developmental toxicity endpoints associated with administration of repeated doses of the material in Wistar rats. The animals were randomly allocated to the four main groups (13/sex/group) and two recovery groups (5/sex/group). The doses selected for main groups were; 0 (G1-control),308 mg/kg body weight (G2), 556 mg/kg body weight (G3) and 1000 mg/kg body weight (G4) daily for 64 days. The recovery groups G1-R and G4-R were dosed with similar doses of respective main groups.Vehicle corn oil to G1 and G1-R and test item to G2, G3, G4 and G4-R animals were administered by oral gavage route each day during the dosing period. No mortality and morbidity were observed any of the groups of animals throughout the study period. Animals of all dose groups were observed for Clinical signs/ symptoms daily once during the experimental period. 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. 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 were comparable and no statistically significant changes were revealed in animals of treatment 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 as compare to the repective control groups. Motor activity measurements were comparable and no changes were revealed in any of the animals of all treated groups of both the sexes as compared to control group. Estrous cycle was evaluated for checking the regularity during treatment period and in cohabitation for confirmation of pregnancy.
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. All hematological and clinical chemistry parameters in animals of different treated groups of both the sexes were comparable to their respective control groups. No treatment related changes were observed in any of the treatment groups. 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 the respective control group rats. External and visceral examination of all male and female rats of control and all treated groups including recovery groups did not reveal any abnormality of pathological significance. 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. Pups that died among the control and treated groups during the course of study, revealed various lesions when examined externally and internally but the observations were not considered treatment related. From the patho-morphological results presented, it is concluded that, the treatment of Methyl Phenyl acetate at 308, 556 and 1000 mg/kg body weight in male and female rats did not affect adversely and no alteration of pathological significance was observed in any of the organs including reproductive organs. 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. Further these observed lesions are common in occurrence in rodents during toxicological studies. Hence, occurrence of these lesions could be considered as spontaneous or incidental in nature and not to be attributed to the administration of the Test Item.
Based on the findings of Repeated Dose Oral Toxicity Study in combination with Reproduction/ Developmental Toxicity of test material in Wistar Rats with 14 days recovery, where in 0, 308, 556 and 1000 mg/kg body weight, doses were tested;No Observed Adverse Effect Level (NOAEL) is considered to be 556 mg/kg bw. When male and female wistar rats were treated with test material orally.
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