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EC number: 202-213-6 | CAS number: 93-04-9
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Repeated dose toxicity: oral
Administrative data
- Endpoint:
- sub-chronic toxicity: oral
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Justification for type of information:
- Data is from a study report
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 020
- Report date:
- 2020
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- other: OECD TG 443: Extended One-Generation Reproductive Toxicity Study
- Principles of method if other than guideline:
- According to OECD TG 443: Extended One-Generation Reproductive Toxicity Study
- GLP compliance:
- yes
- Limit test:
- no
Test material
- Reference substance name:
- Methyl 2-naphthyl ether
- EC Number:
- 202-213-6
- EC Name:
- Methyl 2-naphthyl ether
- Cas Number:
- 93-04-9
- Molecular formula:
- C11H10O
- IUPAC Name:
- 2-methoxynaphthalene
- Test material form:
- solid: crystalline
- Details on test material:
- Name: Methyl-2-naphthyl ether
CAS No.: 93-04-9
IUPAC Name: 2-Methoxynaphthalene
Molecular Weight: 158.19 g/mol
Molecular Formula: C11-H10-O
SMILES: COc1ccc2ccccc2c1
Constituent 1
Test animals
- Species:
- rat
- Strain:
- Wistar
- Details on species / strain selection:
- Rat is the commonly used species for toxicity studies and recommended by the regulatory guidelines and Wistar strain is a commonly used strain to assess the reproduction and developmental toxicity in the reproductive toxicity studies.
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: The test animals were procured from a CPCSEA approved vendor.
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: (P) 12-14 weeks at the start of treatment wks; (F1) 11-12 weeks at the start of treatment wks
- Weight at study initiation: (P) Males: 291-413 g; Females: 198-277 g; (F1) Males: x-x g; Females: x-x g
- Fasting period before study: No Data Available
- Housing: Two to four rats were housed in each polycarbonate cage. During mating, one male and one female rat were housed in a single cage. Pregnant and lactating females were housed 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.
- Use of restrainers for preventing ingestion (if dermal): no
- Diet (e.g. ad libitum): A conventional laboratory pellet diet from approved supplier was available ad libitum.
- Water (e.g. ad libitum): Aquaguard™ filtered drinking water was available ad libitum in regularly cleaned bottles.
- Acclimation period: Animals were acclimatized for at least 5 days prior to test item administration.
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.60 to 24.80°C
- Humidity (%): 34.40 to 66.70 %
- Air changes (per hr): At least 12 changes per hour
- Photoperiod (hrs dark / hrs light): 12 hrs dark / 12 hrs light
IN-LIFE DATES:
From: August 8, 2019
To: March 1, 2020
Administration / exposure
- Route of administration:
- oral: gavage
- Details on route of administration:
- While concentrations of the formulation were fixed for each experimental group, the dose volume for each animal was calculated on the basis of the most recently recorded body weight. Formulation was administered in a single dose through oral gavage at a constant volume to weight ratio of 4 mL/ kg body weight.
- Vehicle:
- corn oil
- Details on oral exposure:
- PREPARATION OF DOSING SOLUTIONS: The vehicle used for dose administration was corn oil. Required quantity for each concentration level of test chemical was weighed and triturated in a mortar-pestle, first without vehicle and then in small volume of the vehicle. After thorough trituration, the test chemical and vehicle were transferred quantitatively into a calibrated measuring cylinder and diluted using vehicle up to the required volume of the formulation, before being transferred to the final container. Formulations were prepared on the same day or one day prior to dose administration and stored at room temperature until usage as the test item stability in solution was proved up to 24 hours. At the time of dosing, dose formulations were kept on a magnetic stirrer for maintaining homogeneity of test formulation. The dose formulation samples [of doses viz; G1 (Control), G2 (Low dose), G3 (Mid dose), and G4 (High dose)] were analyzed for homogeneity and concentration. The dose formulation analysis was carried out once on Day 1 of dose administration to animals, on day 91 and on day 182.
DIET PREPARATION
- Rate of preparation of diet (frequency): No Data Available
- Mixing appropriate amounts with (Type of food): No Data Available
- Storage temperature of food: No Data Available
VEHICLE
- Justification for use and choice of vehicle (if other than water): Corn Oil was selected as a vehicle because the test chemical was found to be insoluble in water whereas corn oil provided adequate suspensibility and the formulation was found stable in this vehicle. Further, corn oil is widely used as vehicle in oral toxicity study and the selected concentration is well below the acceptable limit.
- Concentration in vehicle: 0 mg/kg bw: 0 mg/ml; 62.5 mg/kg bw: 15.63 mg/ml; 125 mg/kg bw: 31.25 mg/ml and 250 mg/kg bw: 62.5 mg/ml
- Amount of vehicle (if gavage): 4 mL/ kg
- Lot/batch no. (if required): Batch No.A1906001, N2191405, N2192016, O21904007
- Purity: No Data Available - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- The analytical method was validated with respect to the parameters covering specificity, linearity, limit of quantification (LOQ), precision (% RSD), Accuracy (% recovery) and Homogeneity. The analytical method was validated with respect to the following parameters as below:
1. Specificity: The result of specificity was reported either no interference observed based on visual comparison or the degree of interference does not contribute more than 20 % peak area of the target analyte at the LOQ Level.
2. Limit of Quantification: The LOQ of the method was the minimum concentration/quantity of a component, which could be quantified precisely and accurately.
The LOQ was carried out at the lowest sensitivity of the device under the same instrumental parameters used for the qualified specificity.
The LOQ of a compound for a specific method was determined by injecting in duplicate the working solution (s) made in suitable solvent fortified with test chemical / reference standard, until it produces a minimum signal to noise ratio ≥ 10:1 for precise quantification.
3. Linearity: The linearity was carried out by preparing and analyzing minimum five linear concentrations of reference standard with at least one determination. The linear calibration curve was established by plotting the analyte peak response against concentration (mg/l). The linearity was given in the form of equation of the resulting curve and correlation coefficient (r). The value of correlation coefficient (r) ≥ 0.99 was considered as an acceptable value.
4. Precision (% RSD), Accuracy (% Recovery) and Homogeneity: Precision, Accuracy was established at the level of test concentration used i.e., at low, mid and high dose level along with control. The active ingredient was diluted using sample diluent and injected onto HPLC using validated HPLC method. Minimum six determinations were made at each fortification level. The homogeneity was performed by analysing Top, Middle and Bottom layers of each fortification level. The active ingredient concentration in each replicate of each fortification level, mean of active ingredient concentration, accuracy (% recovery), SD and % RSD was calculated and reported.
The active ingredient concentration of test item in dose formulation was calculated using the following formula:
Y = bX + a
"AI Concentration (mg/L) = " "Y - a" /"b" " × D"
Where,
Y = Peak area of the sample
a = Intercept
b = Slope of the line
D = Dilution factor (if applicable)
X = A.I. Concentration (mg/L)
The accuracy (% Recovery) will be calculated using the following formula:
"Accuracy " ("% Recovery" )"= " "Obtained Concentration" /"Fortified concentration" " ×100"
The precision (% RSD) will be calculated using the formula provided as below:
"Precision (% RSD)= " "Standard Deviation" /"Mean Recovered Concentration" " ×100"
All the paramaters were according to and within the acceptance criteria of the above test. - Duration of treatment / exposure:
- Dosing in all animals was carried out on a 7-days/week basis from the treatment initiation until necropsy. All animals of both sexes were dosed 2 weeks prior to mating. Dosing was continued in both sexes during the mating period. Females were treated throughout gestation and lactation up to the day of termination after weaning. Males were treated in the same manner until termination. The parental animals were dosed until 7 weeks.
- Frequency of treatment:
- Once Daily
Doses / concentrationsopen allclose all
- Dose / conc.:
- 0 mg/kg bw/day (actual dose received)
- Remarks:
- Control Group (G1)
- Dose / conc.:
- 62.5 mg/kg bw/day (actual dose received)
- Remarks:
- Low Dose Group (G2)
- Dose / conc.:
- 125 mg/kg bw/day (actual dose received)
- Remarks:
- Mid Dose Group (G3)
- Dose / conc.:
- 250 mg/kg bw/day (actual dose received)
- Remarks:
- High Dose Group (G4)
- No. of animals per sex per dose:
- 30 animals per sex per dose
- Control animals:
- yes, concurrent vehicle
- Details on study design:
- - Dose selection rationale:
Doses were selected on the basis of previously performed GLP compliant repeated dose 28 days oral toxicity study. Two deaths were observed in the highest dose (500 mg/kg bw) unrelated to the gavage error of the OECD 407 study and therefore a subsequent lower dose of 250 mg/kg bw was selected as a highest dose in current study.
- Rationale for animal assignment (if not random): During the acclimatisation phase, male and female animals were randomized into four different experimental groups, based on the most recently recorded body weights, using the “Daniel’s XL Toolbar” (http://xltoolbox.sourceforge.net/) for MS Excel. After randomisation, it was ensured that individual body weights were within ± 20% of the respective group means. Details of the randomization were documented in the study raw data. The mean body weights were analysed by One-way ANOVA and it was found that the group means were statistically comparable to each other within each sex and group.
- Fasting period before blood sampling for clinical biochemistry: Animals were overnight fasted prior to blood collection.
- Rationale for selecting satellite groups: No Data Available
- Post-exposure recovery period in satellite groups: No Data Available
- Section schedule rationale (if not random): No Data Available
- Other: No Data Available - Positive control:
- No Data Available
Examinations
- Observations and examinations performed and frequency:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: All animals were observed at least twice daily for cage size observation, throughout the acclimatization and study periods.
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: General clinical observations of rats from all the groups were made once a day, preferably at the same time each day and considering the peak period of anticipated effects after dosing.
BODY WEIGHT: Yes
- Time schedule for examinations: Parental animals were weighed once on receipt, once during randomization, once at the start of treatment and once every week thereafter. In addition, pregnant females were weighed during gestation (GD 0, GD 7, GD 14 and GD 20) and lactation on the same days as the weighing of the pups in their litters.
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes, during the study, feed consumption was recorded weekly or on the same days as animal body weights (except during cohabitation).
- 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 EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: Yes
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Not specified
- Time schedule for examinations:
OPHTHALMOSCOPIC EXAMINATION: No
- Time schedule for examinations: No Data Available
- Dose groups that were examined: No Data Available
HAEMATOLOGY: Yes
- Time schedule for collection of blood: Prior to termination of the parental animals
- Anaesthetic used for blood collection: Yes (isofluorane)
- Animals fasted: Yes
- How many animals: 10 males and 10 females, randomly selected from each group of parental animals
- Parameters checked in table [1.1] were examined.
CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Prior to termination of the parental animals
- Animals fasted: Yes (isofluorane)
- How many animals: 10 males and 10 females, randomly selected from each group of parental animals
- Parameters checked in table [1.2] were examined.
URINALYSIS: Yes
- Time schedule for collection of urine: During the last week of the treatment, 10 males and 10 females, randomly selected from each group of parental animals. During the last week of the treatment, 10 males and 10 females, randomly selected from each group of parental animals.
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes
- Parameters checked in table [1.3] were examined.
NEUROBEHAVIOURAL EXAMINATION: No
- Time schedule for examinations: No Data Available
- Dose groups that were examined: No Data Available
- Battery of functions tested: No Data Available
IMMUNOLOGY: Not specified
- Time schedule for examinations: No Data Available
- How many animals: No Data Available
- Dose groups that were examined: No Data Available
OTHER: Hormone Analysis: From 10 males and 10 females, randomly selected from each group of parental animals , blood was collected in the morning; serum was separated and stored under appropriate conditions (-70°C ± 5°C). All collected blood samples were assessed for serum levels of T4, TSH, Estradiol, Progesterone and Testosterone using commercially available Rat ELISA kits. - Sacrifice and pathology:
- GROSS PATHOLOGY: Yes (see table 1.4)
HISTOPATHOLOGY: Yes (see table 1.4), full histopathology was carried out on the preserved organs and tissues of all parental animals in the control and high dose group of both sexes. Slides for low- and mid-dose groups were processed only if, test-chemical related pathologies were observed in the high-dose group sections. - Other examinations:
- Organ Weights: At the time of termination, organs marked with asterisk (*) in table 1.4 from all parental animals were trimmed of adherent tissue/ fat and weighed, prior to preservation in fixative.
- Statistics:
- The findings of this study were evaluated in terms of the observed effects, necropsy and microscopic findings. The evaluation will included the relationship between the dose of the test substance and the presence or absence, the incidence and severity, of abnormalities. Raw data was processed using statistical software. The mean and standard deviation was calculated using the software and all data was summarized in tabular form. All continuous data were checked for their homogeneity using Shapiro Wilk test. All homogenous data were analyzed using ANOVA and data showing significance in their variances were subjected to Dunnett’s t-test. All heterogeneous data was analysed using Kruskal-Wallis ANOVA on ranks and data showing significance in their variances were subjected to Dunn’s Test. P values of less than or equal to 0.05 were accepted as being statistically significant.
Results and discussion
Results of examinations
- Clinical signs:
- effects observed, treatment-related
- Description (incidence and severity):
- No treatment-related clinical signs or symptoms were observed in any of the parent animals up to 125 mg/kg body weight of the test chemical. However, mild salivation was observed in total 13 of 30 males of G4 group (250 mg/kg), starting from the 27th day of treatment and continuing till termination. All adult animals were found normal with respect to all the parameters examined during detailed clinical examinations through the course of the study. No treatment-related symptoms were found up to the dose level of 250 mg/kg body weight in either sex.
- Mortality:
- no mortality observed
- Description (incidence):
- No morbidities or mortality were recorded in adult animals of parent generation in any of the experimental groups throughout the duration of the experiment.
- Body weight and weight changes:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Mean body weights remained comparable among all the groups for every time point (days 8, 15, 22, 29, 36, 43, 50, 57, 64 and 71) recorded during the 70 days exposure period for males. No significant difference was found for the mean body weights of females during the two weeks pre-mating period, during mating, during gestation or during lactation period. However, the mean percent change in body weight on all the instances (days 8, 15, 22, 29, 36, 43, 50, 57, 64 and 71) with respect to the first day of exposure was significantly reduced for parental males of G4 group (250 mg/kg) (P≤0.05 to P≤0.001) as compared to control animals, whereas mean body weight change for G2 (62.5 mg/kg) and G3 (125 mg/kg) males remained comparable to control animals. Among the females, mean percent change in body weights remained statistically comparable among all groups during pre-mating, mating, gestation and lactation periods.
- Food consumption and compound intake (if feeding study):
- effects observed, non-treatment-related
- Description (incidence and severity):
- The mean feed consumed per animal per day significantly increased on day 8 of treatment for G2 (62.5 mg/kg) (P≤0.01) and G3 (125 mg/kg) (P≤0.05) males as compared to G1 (control) animals. However, it was found to be similar for all other time points across the groups. The mean feed consumption of treatment group (G2, G3 & G4) females remained statistically comparable to control group during pre-mating, mating, gestation and lactation period except for one instance, where mean feed consumption on day 8 of treatment was significantly reduced for G2 (62.5 mg/kg) (P≤0.01) as compared to control females. The differences found in mean feed consumption of male and female animals were neither dose-dependent nor consistent, hence it was deemed to be independent of test-chemical treatment.
- Food efficiency:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Feed efficiency for the G4 males had statistically reduced in the first two weeks of treatment, i.e. on days 8 and 15. It was also found to have increased significantly during the penultimate week of treatment, i.e. on day 63. For all other groups among the males, the feed efficiency was statistically comparable across the groups. Among the parental females, no statistical differences were observed for feed efficiency in any of the groups.
- 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):
- No significant difference was observed in any of the measured haematology parameters in all the groups of both the sexes, except in the WBC count, which was found to be significantly increased in males of G4 group (P≤0.01) as compared to control males.
- Clinical biochemistry findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Significant increase was observed in Phosphorus, Sodium and Chloride levels in G4 males (P≤0.05) as compared to control males. All the other clinical chemistry parameters in males and females were found comparable in control and treatment groups.
- Urinalysis findings:
- no effects observed
- Description (incidence and severity):
- The urine parameters tested did not show up any significant changes among the experimental groups in the parental generation of either sex.
- Behaviour (functional findings):
- not specified
- Immunological findings:
- not specified
- Organ weight findings including organ / body weight ratios:
- effects observed, treatment-related
- Description (incidence and severity):
- No significant difference was observed in absolute organ weight or organ weight relative to body weight for the entire set of measured organs in parent male and female animals. However, significant reduction was observed in absolute and relative liver weight in G4 females (P<0.05) as compared to control animals. In the current study, however, the decreased liver weights with respect to body weights in G4 females does not correlate with any histological observations made, making it inconsequential from the point of view of toxic effects of test-chemical.
- Gross pathological findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- External examination of all male and female rats of control and all treated groups did not reveal any lesion of pathological significance. Internal examination of the rats revealed reduced testicular size in two animals (Male: G2: 1/30, G4:1/30), increased prostate size in one animal (Male: G2: 1/30) and reduced prostate size in one animal (Male: G3: 1/30). Remaining animals of parent generation did not reveal any abnormality of pathological significance.
- Neuropathological findings:
- not specified
- Histopathological findings: non-neoplastic:
- effects observed, non-treatment-related
- Description (incidence and severity):
- No treatment-related histopathological findings are reported in this study that could arise out of test-item administration. All observed tissues were normal at the level of histology. Minimal to moderate and focal lymphocytic infiltrates were observed in some slides of liver, kidney, lung, heart, colon, thymus and adrenals with mild to moderate cyst in thyroid gland, which were distributed randomly across the experimental groups and did not show any pattern of either dose-dependency or sex-based selectivity.
- Histopathological findings: neoplastic:
- not specified
- Other effects:
- effects observed, treatment-related
- Description (incidence and severity):
- While TSH, Estradiol, Progesterone and Testosterone remained statistically comparable among the groups of parental males, the mean level of T4 was found reduced in G3 males (P<0.01) as compared to control. This is likely an incidental result, given that G4 was statistically comparable to control. The mean T4, TSH, Estradiol and testosterone levels remained comparable among the groups of parental females; whereas, Progesterone was found to be significantly reduced in G4 females (P<0.001). This could be a test-chemical related effect.
- Details on results:
- No treatment-related clinical signs or symptoms were observed in any of the parent animals up to 125 mg/kg body weight of the test chemical. However, mild salivation was observed in total 13 of 30 males of G4 group (250 mg/kg), starting from the 27th day of treatment and continuing till termination. All adult animals were found normal with respect to all the parameters examined during detailed clinical examinations through the course of the study. No treatment-related symptoms were found up to the dose level of 250 mg/kg body weight in either sex. No morbidities or mortality were recorded in adult animals of parent generation in any of the experimental groups throughout the duration of the experiment. Mean body weights remained comparable among all the groups for every time point (days 8, 15, 22, 29, 36, 43, 50, 57, 64 and 71) recorded during the 70 days exposure period for males. No significant difference was found for the mean body weights of females during the two weeks pre-mating period, during mating, during gestation or during lactation period. However, the mean percent change in body weight on all the instances (days 8, 15, 22, 29, 36, 43, 50, 57, 64 and 71) with respect to the first day of exposure was significantly reduced for parental males of G4 group (250 mg/kg) (P≤0.05 to P≤0.001) as compared to control animals, whereas mean body weight change for G2 (62.5 mg/kg) and G3 (125 mg/kg) males remained comparable to control animals. Among the females, mean percent change in body weights remained statistically comparable among all groups during pre-mating, mating, gestation and lactation periods. The mean feed consumed per animal per day significantly increased on day 8 of treatment for G2 (62.5 mg/kg) (P≤0.01) and G3 (125 mg/kg) (P≤0.05) males as compared to G1 (control) animals. However, it was found to be similar for all other time points across the groups. The mean feed consumption of treatment group (G2, G3 & G4) females remained statistically comparable to control group during pre-mating, mating, gestation and lactation period except for one instance, where mean feed consumption on day 8 of treatment was significantly reduced for G2 (62.5 mg/kg) (P≤0.01) as compared to control females. The differences found in mean feed consumption of male and female animals were neither dose-dependent nor consistent, hence it was deemed to be independent of test-item treatment. Feed efficiency for the G4 males had statistically reduced in the first two weeks of treatment, i.e. on days 8 and 15. It was also found to have increased significantly during the penultimate week of treatment, i.e. on day 63. For all other groups among the males, the feed efficiency was statistically comparable across the groups. Among the parental females, no statistical differences were observed for feed efficiency in any of the groups. No significant difference was observed in any of the measured haematology parameters in all the groups of both the sexes, except in the WBC count, which was found to be significantly increased in males of G4 group (P≤0.01) as compared to control males. Significant increase was observed in Phosphorus, Sodium and Chloride levels in G4 males (P≤0.05) as compared to control males. All the other clinical chemistry parameters in males and females were found comparable in control and treatment groups. The urine parameters tested did not show up any significant changes among the experimental groups in the parental generation of either sex. No significant difference was observed in absolute organ weight or organ weight relative to body weight for the entire set of measured organs in parent male and female animals. However, significant reduction was observed in absolute and relative liver weight in G4 females (P<0.05) as compared to control animals. In the current study, however, the decreased liver weights with respect to body weights in G4 females does not correlate with any histological observations made, making it inconsequential from the point of view of toxic effects of test-chemical. External examination of all male and female rats of control and all treated groups did not reveal any lesion of pathological significance. Internal examination of the rats revealed reduced testicular size in two animals (Male: G2: 1/30, G4:1/30), increased prostate size in one animal (Male: G2: 1/30) and reduced prostate size in one animal (Male: G3: 1/30). Remaining animals of parent generation did not reveal any abnormality of pathological significance. No treatment-related histopathological findings are reported in this study that could arise out of test-item administration. All observed tissues were normal at the level of histology. Minimal to moderate and focal lymphocytic infiltrates were observed in some slides of liver, kidney, lung, heart, colon, thymus and adrenals with mild to moderate cyst in thyroid gland, which were distributed randomly across the experimental groups and did not show any pattern of either dose-dependency or sex-based selectivity. While TSH, Estradiol, Progesterone and Testosterone remained statistically comparable among the groups of parental males, the mean level of T4 was found reduced in G3 males (P<0.01) as compared to control. This is likely an incidental result, given that G4 was statistically comparable to control. The mean T4, TSH, Estradiol and testosterone levels remained comparable among the groups of parental females; whereas, Progesterone was found to be significantly reduced in G4 females (P<0.001). This could be a test-chemical related effect.
Effect levels
open allclose all
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 125 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- body weight and weight gain
- clinical biochemistry
- clinical signs
- food consumption and compound intake
- food efficiency
- gross pathology
- haematology
- histopathology: non-neoplastic
- mortality
- organ weights and organ / body weight ratios
- urinalysis
- other: Hormone Analysis
- Remarks on result:
- not determinable due to absence of adverse toxic effects
- Dose descriptor:
- LOAEL
- Effect level:
- 250 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- clinical signs
- organ weights and organ / body weight ratios
- other: Hormone Analysis
- Remarks on result:
- other: Please see 'remarks'
- Remarks:
- There were treatment related changes observed during clinical signs examination, organ weights and progesterone (hormonal) levels.
Target system / organ toxicity
- Critical effects observed:
- no
Any other information on results incl. tables
Table 2.1: Mortality and Morbidity Record
|
Sex: Male |
|||
Group |
Dose (mg/kg) |
No. of Animals |
Observation |
|
G1 |
0 |
30 |
No mortality or morbidity observed |
|
G2 |
62.5 |
30 |
No mortality or morbidity observed |
|
G3 |
125 |
30 |
No mortality or morbidity observed |
|
G4 |
250 |
30 |
No mortality or morbidity observed |
|
|
Sex: Female |
|||
Group |
Dose (mg/kg) |
No. of Animals |
Observation |
|
G1 |
0 |
30 |
No mortality or morbidity observed |
|
G2 |
62.5 |
30 |
No mortality or morbidity observed |
|
G3 |
125 |
30 |
No mortality or morbidity observed |
|
G4 |
250 |
30 |
No mortality or morbidity observed |
|
Table 2.2: Clinical Signs and Symptoms Record
|
Sex: Male |
|||
Group |
Dose (mg/kg) |
No. of Animals |
Observation |
|
G1 |
0 |
30 |
Normal |
|
G2 |
62.5 |
30 |
Normal |
|
G3 |
125 |
30 |
Normal |
|
G4 |
250 |
30 |
Normal |
|
|
Sex: Female |
|||
Group |
Dose (mg/kg) |
No. of Animals |
Observation |
|
G1 |
0 |
30 |
Normal |
|
G2 |
62.5 |
30 |
Normal |
|
G3 |
125 |
30 |
Normal |
|
G4 |
250 |
30 |
*Normal |
Table 2.3 Mean Body weights (g)
Sex: Male |
||||||||||||||
Group |
Dose (mg/Kg) |
|
Day of Exposure |
|||||||||||
1 |
8 |
15 |
22 |
29 |
36 |
43 |
50 |
57 |
64 |
71 |
||||
G1 |
0 |
Mean |
351.73 |
359.60 |
371.80 |
384.63 |
400.93 |
419.17 |
426.90 |
431.53 |
441.13 |
444.70 |
452.53 |
|
SD |
26.71 |
26.09 |
25.43 |
27.51 |
28.22 |
30.60 |
31.14 |
30.71 |
29.88 |
33.09 |
32.31 |
|||
N |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
|||
G2 |
62.5 |
Mean |
357.10 |
365.17 |
377.03 |
387.97 |
406.10 |
426.53 |
434.03 |
439.53 |
448.87 |
451.17 |
459.23 |
|
SD |
26.62 |
24.64 |
22.51 |
23.92 |
26.57 |
28.79 |
29.40 |
29.89 |
29.65 |
31.22 |
32.62 |
|||
N |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
|||
G3 |
125 |
Mean |
356.17 |
363.80 |
373.40 |
385.77 |
402.57 |
419.73 |
428.73 |
435.50 |
441.70 |
447.23 |
454.03 |
|
SD |
25.19 |
24.28 |
24.93 |
28.35 |
26.99 |
30.13 |
33.92 |
32.82 |
31.43 |
34.62 |
38.82 |
|||
N |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
|||
G4 |
250 |
Mean |
356.17 |
359.17 |
365.40 |
377.27 |
392.43 |
407.17 |
413.50 |
417.67 |
427.13 |
435.77 |
441.33 |
|
SD |
26.31 |
25.80 |
25.49 |
26.75 |
31.22 |
31.45 |
32.22 |
34.58 |
39.78 |
41.59 |
42.78 |
|||
N |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
|||
Pre-mating |
Sex: Female |
||||||
Group |
Dose (mg/Kg) |
|
Day of Exposure |
||||
1 |
8 |
15 |
22 |
||||
G1 |
0 |
Mean |
238.07 |
242.10 |
246.67 |
256.50 |
|
SD |
17.01 |
16.63 |
16.35 |
14.89 |
|||
N |
30 |
30 |
30 |
4 |
|||
G2 |
62.5 |
Mean |
237.60 |
242.53 |
248.17 |
272.50 |
|
SD |
20.80 |
20.77 |
19.25 |
16.26 |
|||
N |
30 |
30 |
30 |
2 |
|||
G3 |
125 |
Mean |
241.83 |
244.50 |
247.47 |
244.25 |
|
SD |
19.86 |
20.38 |
21.86 |
14.45 |
|||
N |
30 |
30 |
30 |
4 |
|||
G4 |
250 |
Mean |
240.87 |
241.93 |
245.80 |
258.00 |
|
SD |
16.30 |
15.80 |
14.70 |
22.63 |
|||
N |
30 |
30 |
30 |
2 |
|||
Keys:N: number; SD: standard deviation
Gestation |
Sex: Female |
||||||
Group |
Dose (mg/Kg) |
|
Day of Gestation |
||||
0 |
7 |
14 |
20 |
||||
G1 |
0 |
Mean |
253.72 |
280.45 |
310.76 |
374.66 |
|
SD |
17.43 |
19.74 |
22.43 |
29.95 |
|||
N |
29 |
29 |
29 |
29 |
|||
G2 |
62.5 |
Mean |
253.36 |
282.14 |
312.32 |
372.75 |
|
SD |
21.86 |
23.49 |
24.03 |
30.58 |
|||
N |
28 |
28 |
28 |
28 |
|||
G3 |
125 |
Mean |
250.93 |
278.55 |
310.45 |
372.38 |
|
SD |
20.57 |
20.32 |
24.60 |
30.02 |
|||
N |
29 |
29 |
29 |
29 |
|||
G4 |
250 |
Mean |
247.04 |
275.42 |
306.04 |
364.27 |
|
SD |
15.24 |
17.44 |
20.50 |
27.77 |
|||
N |
26 |
26 |
26 |
26 |
|||
Lactation |
Sex: Female |
|||||||
Group |
Dose (mg/Kg) |
|
Day of Lactation |
|||||
0 |
4 |
7 |
14 |
21 |
||||
G1 |
0 |
Mean |
293.83 |
294.10 |
294.90 |
299.31 |
296.38 |
|
SD |
19.69 |
21.02 |
21.57 |
25.65 |
23.89 |
|||
N |
29 |
29 |
29 |
29 |
29 |
|||
G2 |
62.5 |
Mean |
295.23 |
300.73 |
303.13 |
308.03 |
307.90 |
|
SD |
29.28 |
24.37 |
22.64 |
24.24 |
25.74 |
|||
N |
28 |
28 |
28 |
28 |
28 |
|||
G3 |
125 |
Mean |
293.48 |
293.21 |
297.86 |
305.03 |
304.83 |
|
SD |
27.80 |
26.55 |
25.42 |
27.10 |
26.87 |
|||
N |
29 |
29 |
29 |
29 |
29 |
|||
G4 |
250 |
Mean |
293.88 |
292.58 |
299.12 |
304.77 |
302.19 |
|
SD |
26.71 |
27.01 |
28.33 |
26.21 |
26.98 |
|||
N |
26 |
26 |
26 |
26 |
26 |
|||
Keys:N: number; SD: standard deviation
Non-Parturated |
Sex: Female |
|||||||||
Group |
Dose (mg/Kg) |
|
Day of Gestation |
|||||||
0 |
7 |
14 |
20 |
27 |
30 |
34 |
||||
G1 |
0 |
Mean |
260.00 |
261.00 |
271.00 |
274.00 |
271.00 |
./. |
270.00 |
|
SD |
./. |
./. |
./. |
./. |
./. |
./. |
./. |
|||
N |
1 |
1 |
1 |
1 |
1 |
0 |
1 |
|||
G2 |
62.5 |
Mean |
271.50 |
274.50 |
279.50 |
278.50 |
281.50 |
289.00 |
264.00 |
|
SD |
41.72 |
37.48 |
12.02 |
16.26 |
21.92 |
./. |
./. |
|||
N |
2 |
2 |
2 |
2 |
2 |
1 |
1 |
|||
G3 |
125 |
Mean |
251.00 |
252.00 |
257.00 |
255.00 |
255.00 |
./. |
258.00 |
|
SD |
./. |
./. |
./. |
./. |
./. |
./. |
./. |
|||
N |
1 |
1 |
1 |
1 |
1 |
0 |
1 |
|||
G4 |
250 |
Mean |
254.25 |
268.50 |
272.25 |
270.50 |
274.00 |
./. |
271.00 |
|
SD |
17.69 |
12.66 |
20.85 |
20.44 |
21.23 |
./. |
20.85 |
|||
N |
4 |
4 |
4 |
4 |
4 |
0 |
4 |
|||
Keys:N: number; SD: standard deviation;./.= Not applicable
Table 2.4 Mean Body weight Change (%)
Sex: Male |
|||||||||||||||||||
Group |
Dose (mg/Kg) |
|
Day of Exposure |
||||||||||||||||
8 |
15 |
22 |
29 |
36 |
43 |
50 |
57 |
64 |
71 |
|
|||||||||
G1 |
0 |
Mean |
2.28 |
5.81 |
9.46 |
14.11 |
19.26 |
21.47 |
22.83 |
25.59 |
26.57 |
28.83 |
|
||||||
SD |
2.19 |
3.17 |
3.81 |
4.23 |
3.92 |
4.07 |
4.89 |
4.99 |
5.55 |
5.66 |
|
||||||||
N |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
|
||||||||
G2 |
62.5 |
Mean |
2.34 |
5.75 |
8.79 |
13.84 |
19.57 |
21.69 |
23.23 |
25.88 |
26.49 |
28.74 |
|
||||||
SD |
2.39 |
3.62 |
3.29 |
3.39 |
4.00 |
4.62 |
4.85 |
5.24 |
5.26 |
5.44 |
|
||||||||
N |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
|
||||||||
G3 |
125 |
Mean |
2.20 |
4.92 |
8.35 |
13.13 |
17.89 |
20.38 |
22.32 |
24.10 |
25.62 |
27.49 |
|
||||||
SD |
2.35 |
3.59 |
3.79 |
4.14 |
3.85 |
4.37 |
4.57 |
4.89 |
5.22 |
6.30 |
|
||||||||
N |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
|
||||||||
G4 |
250 |
Mean |
0.87↓ |
2.65↓↓↓ |
5.99↓↓↓ |
10.20↓↓↓ |
14.38↓↓↓ |
16.17↓↓↓ |
17.32↓↓↓ |
19.96↓↓↓ |
22.36↓↓ |
23.93↓↓ |
|
||||||
SD |
1.38 |
2.43 |
2.86 |
3.68 |
4.38 |
4.73 |
5.21 |
7.03 |
7.22 |
7.63 |
|
||||||||
N |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
|
||||||||
Pre-mating |
Sex: Female |
|
|||||||||||||||||
Group |
Dose (mg/Kg) |
|
Day of Exposure |
|
|||||||||||||||
8 |
15 |
22 |
|
||||||||||||||||
G1 |
0 |
Mean |
1.73 |
3.71 |
8.14 |
|
|||||||||||||
SD |
2.12 |
3.96 |
2.73 |
|
|||||||||||||||
N |
30 |
30 |
4 |
|
|||||||||||||||
G2 |
62.5 |
Mean |
2.12 |
4.60 |
11.64 |
|
|||||||||||||
SD |
2.46 |
4.08 |
4.00 |
|
|||||||||||||||
N |
30 |
30 |
2 |
|
|||||||||||||||
G3 |
125 |
Mean |
1.11 |
2.33 |
7.00 |
|
|||||||||||||
SD |
1.84 |
3.07 |
4.96 |
|
|||||||||||||||
N |
30 |
30 |
4 |
|
|||||||||||||||
G4 |
250 |
Mean |
0.46 |
2.12 |
5.38 |
|
|||||||||||||
SD |
1.05 |
2.25 |
4.44 |
|
|||||||||||||||
N |
30 |
30 |
2 |
|
Key: Body weight change (%) with respect to Day 1 of treatment; ↓: decreased as compared to control (P<0.05); ↓↓: decreased as compared to control (P<0.01); ↓↓↓: decreased as compared to control (P<0.001)
Gestation |
Sex: Female |
|||||
Group |
Dose (mg/Kg) |
|
Day of Gestation |
|||
7 |
14 |
20 |
||||
G1 |
0 |
Mean |
10.63 |
22.69 |
47.92 |
|
SD |
5.04 |
7.67 |
10.38 |
|||
N |
29 |
29 |
29 |
|||
G2 |
62.5 |
Mean |
11.42 |
23.41 |
47.30 |
|
SD |
2.70 |
3.34 |
6.02 |
|||
N |
28 |
28 |
28 |
|||
G3 |
125 |
Mean |
11.14 |
23.83 |
48.53 |
|
SD |
3.65 |
4.62 |
6.04 |
|||
N |
29 |
29 |
29 |
|||
G4 |
250 |
Mean |
11.50 |
23.92 |
47.47 |
|
SD |
2.09 |
4.45 |
7.00 |
|||
N |
26 |
26 |
26 |
Key: Body weight change (%) with respect to Day 0 of Gestation
Lactation |
Sex: Female |
||||||
Group |
Dose (mg/Kg) |
|
Day of Lactation |
||||
4 |
7 |
14 |
21 |
||||
G1 |
0 |
Mean |
0.18 |
0.46 |
1.95 |
1.07 |
|
SD |
4.9 |
5.5 |
6.85 |
7.89 |
|||
N |
29 |
29 |
29 |
29 |
|||
G2 |
62.5 |
Mean |
2.15 |
3.04 |
4.73 |
4.67 |
|
SD |
4.83 |
5.44 |
6.86 |
6.86 |
|||
N |
28 |
28 |
28 |
28 |
|||
G3 |
125 |
Mean |
0.04 |
1.71 |
4.17 |
4.09 |
|
SD |
4.35 |
5.34 |
6.45 |
6.2 |
|||
N |
29 |
29 |
29 |
29 |
|||
G4 |
250 |
Mean |
-0.33 |
1.93 |
3.97 |
3.22 |
|
SD |
5.12 |
6.24 |
6.91 |
8.61 |
|||
N |
26 |
26 |
26 |
26 |
Key: Body weight change (%) with respect to Day 0 of Lactation
Non-Parturated |
Sex: Female |
||||||||
Group |
Dose (mg/Kg) |
|
Day of Gestation |
||||||
7 |
14 |
20 |
27 |
30 |
34 |
||||
G1 |
0 |
Mean |
0.38 |
4.23 |
5.38 |
4.23 |
./. |
3.85 |
|
SD |
./. |
./. |
./. |
./. |
./. |
./. |
|||
N |
1 |
1 |
1 |
1 |
0 |
1 |
|||
G2 |
62.5 |
Mean |
1.24 |
3.83 |
3.34 |
4.29 |
-3.99 |
9.09 |
|
SD |
1.75 |
11.53 |
9.89 |
7.95 |
./. |
./. |
|||
N |
2 |
2 |
2 |
2 |
1 |
1 |
|||
G3 |
125 |
Mean |
0.40 |
2.39 |
1.59 |
1.59 |
./. |
2.79 |
|
SD |
./. |
./. |
./. |
./. |
./. |
./. |
|||
N |
1 |
1 |
1 |
0 |
1 |
0 |
|||
G4 |
250 |
Mean |
5.74 |
7.41 |
6.49 |
7.76 |
./. |
6.61 |
|
SD |
3.00 |
10.47 |
5.99 |
3.05 |
./. |
4.03 |
|||
N |
4 |
4 |
4 |
4 |
0 |
4 |
Table 2.5 Summary of Feed Consumption (gram/animal/day)
Sex: Male |
||||||||||||
Group |
Dose (mg/Kg) |
|
Day of Exposure |
|||||||||
1 to 8 |
8 to 15 |
28 to 36 |
36 to 43 |
43 to50 |
50 to 57 |
57 to 64 |
64 to 71 |
|||||
G1 |
0 |
Mean |
15.82 |
20.01 |
19.78 |
19.10 |
19.74 |
19.57 |
20.54 |
21.00 |
||
SD |
2.90 |
1.52 |
1.66 |
1.33 |
1.47 |
1.10 |
1.78 |
1.58 |
||||
N |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
||||
G2 |
62.5 |
Mean |
19.09↑↑ |
19.36 |
20.10 |
19.36 |
19.88 |
19.81 |
20.04 |
21.13 |
||
SD |
1.89 |
1.59 |
1.30 |
1.27 |
1.48 |
1.23 |
1.92 |
1.55 |
||||
N |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
||||
G3 |
125 |
Mean |
18.73↑ |
19.18 |
19.71 |
19.32 |
20.31 |
20.31 |
21.37 |
21.90 |
||
SD |
1.48 |
1.73 |
1.48 |
1.53 |
1.78 |
1.23 |
2.11 |
2.10 |
||||
N |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
||||
G4 |
250 |
Mean |
18.03 |
18.88 |
19.04 |
19.07 |
19.59 |
20.02 |
20.91 |
20.08 |
||
SD |
1.39 |
1.42 |
1.73 |
1.28 |
1.33 |
1.76 |
1.83 |
2.95 |
||||
N |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
Pre-mating |
Sex: Female |
|
|||||
Group |
Dose (mg/Kg) |
|
Day of Exposure |
||||
1 to 8 |
8 to 15 |
||||||
G1 |
0 |
Mean |
15.00 |
13.89 |
|||
SD |
1.42 |
1.26 |
|||||
N |
30 |
30 |
|||||
G2 |
62.5 |
Mean |
13.20↓↓ |
13.88 |
|||
SD |
2.04 |
1.72 |
|||||
N |
30 |
30 |
|||||
G3 |
125 |
Mean |
14.14 |
14.61 |
|||
SD |
1.47 |
1.56 |
|||||
N |
30 |
30 |
|||||
G4 |
250 |
Mean |
13.69 |
13.68 |
|||
SD |
0.76 |
1.33 |
|||||
N |
30 |
30 |
Key: SD: Standard Deviation; N: Number of Animals; ↑: increased as compared to control (P<0.05); ↑↑: increased as compared to control (P<0.01); ↓↓: decreased as compared to control (P<0.01).
Gestation |
Sex: Female |
|||||||
Group |
Dose (mg/Kg) |
|
Day of Gestation |
|||||
0 to 7 |
7 to 14 |
14 to 20 |
20 to 22 |
20 to 23 |
||||
G1 |
0 |
Mean |
16.71 |
19.13 |
22.30 |
25.43 |
23.00 |
|
SD |
2.57 |
3.95 |
3.35 |
9.27 |
./. |
|||
N |
29 |
29 |
29 |
28 |
1 |
|||
G2 |
62.5 |
Mean |
15.81 |
19.32 |
22.10 |
21.83 |
18.00 |
|
SD |
2.50 |
1.49 |
3.84 |
7.37 |
1.41 |
|||
N |
28 |
28 |
28 |
26 |
2 |
|||
G3 |
125 |
Mean |
16.93 |
19.68 |
21.95 |
26.39 |
17.33 |
|
SD |
1.97 |
1.98 |
3.13 |
11.30 |
1.41 |
|||
N |
29 |
29 |
29 |
27 |
2 |
|||
G4 |
250 |
Mean |
16.30 |
19.15 |
20.62 |
25.65 |
./. |
|
SD |
2.01 |
1.76 |
3.19 |
7.90 |
./. |
|||
N |
26 |
26 |
26 |
26 |
0 |
Lactation |
Sex: Female |
||||||
Group |
Dose (mg/Kg) |
|
Day of Lactation |
||||
0 to 4 |
4 to 7 |
7 to 14 |
14 to 21 |
||||
G1 |
0 |
Mean |
29.47 |
38.51 |
48.31 |
56.82 |
|
SD |
9.17 |
9.11 |
9.07 |
9.02 |
|||
N |
29 |
29 |
29 |
29 |
|||
G2 |
62.5 |
Mean |
27.44 |
39.04 |
47.22 |
57.88 |
|
SD |
5.28 |
7.85 |
10.01 |
6.89 |
|||
N |
29 |
29 |
29 |
29 |
|||
G3 |
125 |
Mean |
26.88 |
40.29 |
47.83 |
57.73 |
|
SD |
5.33 |
10.03 |
7.88 |
9.19 |
|||
N |
29 |
29 |
29 |
29 |
|||
G4 |
250 |
Mean |
24.72 |
35.53 |
47.18 |
55.68 |
|
SD |
6.47 |
8.32 |
6.59 |
10.46 |
|||
N |
26 |
26 |
26 |
26 |
Key: SD: Standard Deviation; N: Number of Animals
Non-Parturated |
Sex: Female |
||||||||
Group |
Dose (mg/Kg) |
|
Day of Gestation |
||||||
0 to 7 |
7 to 14 |
14 to 20 |
20 to 27 |
27 to 30 |
27 to 34 |
||||
G1 |
0 |
Mean |
14.86 |
16.29 |
18.5 |
17.71 |
./. |
17.29 |
|
SD |
./. |
./. |
./. |
./. |
./. |
./. |
|||
N |
1 |
1 |
1 |
1 |
./. |
1 |
|||
G2 |
62.5 |
Mean |
15.22 |
12.5 |
20.42 |
10.5 |
14 |
11.57 |
|
SD |
7.38 |
3.34 |
5.3 |
9.8 |
./. |
./. |
|||
N |
2 |
2 |
2 |
2 |
1 |
1 |
|||
G3 |
125 |
Mean |
10.43 |
12.57 |
12.83 |
11.86 |
./. |
11.57 |
|
SD |
./. |
./. |
./. |
./. |
./. |
./. |
|||
N |
1 |
1 |
1 |
1 |
./. |
1 |
|||
G4 |
250 |
Mean |
15.46 |
15.79 |
13.96 |
18.54 |
./. |
16 |
|
SD |
1.96 |
1.97 |
2.87 |
2.94 |
./. |
1.65 |
|||
N |
4 |
4 |
4 |
4 |
./. |
4 |
Key: SD: Standard Deviation; N: Number of Animals; ./. Not applicable
Table 2.6 Feed efficiency
Sex: Male |
|||||||||||
Group |
Dose (mg/kg) |
|
Feed Efficiency |
||||||||
8 |
15 |
36 |
43 |
50 |
57 |
64 |
71 |
||||
G1 |
0 |
Mean |
0.074 |
0.088 |
0.132 |
0.057 |
0.033 |
0.071 |
0.023 |
0.053 |
|
SD |
0.067 |
0.049 |
0.046 |
0.043 |
0.047 |
0.047 |
0.059 |
0.031 |
|||
N |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
|||
G2 |
62.5 |
Mean |
0.061 |
0.086 |
0.144 |
0.055 |
0.039 |
0.067 |
0.015 |
0.054 |
|
SD |
0.063 |
0.055 |
0.047 |
0.043 |
0.039 |
0.040 |
0.053 |
0.040 |
|||
N |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
|||
G3 |
125 |
Mean |
0.058 |
0.071 |
0.122 |
0.065 |
0.049 |
0.044 |
0.036 |
0.044 |
|
SD |
0.06 |
0.051 |
0.087 |
0.061 |
0.037 |
0.036 |
0.046 |
0.066 |
|||
N |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
|||
G4 |
250 |
Mean |
0.023↓↓ |
0.047↓↓ |
0.110 |
0.047 |
0.030 |
0.064 |
0.059↑ |
0.039 |
|
SD |
0.037 |
0.035 |
0.066 |
0.044 |
0.038 |
0.088 |
0.057 |
0.061 |
|||
N |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
Sex: Female |
|||||
Group |
Dose (mg/kg) |
|
Feed Efficiency |
||
0 |
15 |
||||
G1 |
0 |
Mean |
0.039 |
0.047 |
|
SD |
0.047 |
0.056 |
|||
N |
30 |
30 |
|||
G2 |
62.5 |
Mean |
0.057 |
0.059 |
|
SD |
0.070 |
0.057 |
|||
N |
30 |
30 |
|||
G3 |
125 |
Mean |
0.026 |
0.028 |
|
SD |
0.044 |
0.035 |
|||
N |
30 |
30 |
|||
G4 |
250 |
Mean |
0.011 |
0.039 |
|
SD |
0.026 |
0.037 |
|||
N |
30 |
30 |
Key: SD: Standard Deviation; N: Number of Animals;↑: increased as compared to control (P<0.05);↓↓: decreased as compared to control (P<0.01).
Gestation |
Sex: Female |
||||||
Group |
Dose (mg/kg) |
|
Feed Efficiency |
||||
0 |
15 |
20 |
|||||
G1 |
0 |
Mean |
0.229 |
0.209 |
0.480 |
||
SD |
0.102 |
0.127 |
0.070 |
||||
N |
30 |
30 |
30 |
||||
G2 |
62.5 |
Mean |
0.264 |
0.223 |
0.465 |
||
SD |
0.063 |
0.054 |
0.095 |
||||
N |
28 |
28 |
28 |
||||
G3 |
125 |
Mean |
0.236 |
0.231 |
0.478 |
||
SD |
0.078 |
0.058 |
0.084 |
||||
N |
28 |
28 |
28 |
||||
G4 |
250 |
Mean |
0.251 |
0.226 |
0.476 |
||
SD |
0.048 |
0.066 |
0.093 |
||||
N |
26 |
26 |
26 |
Non Parturated Dam |
Sex: Female |
|||||||
Group |
Dose (mg/kg) |
Feed Efficiency |
||||||
0 |
15 |
20 |
27 |
|||||
G1 |
0 |
Mean |
0.010 |
0.088 |
0.027 |
-0.024 |
||
SD |
./. |
./. |
./. |
./. |
||||
N |
1 |
1 |
1 |
1 |
||||
G2 |
62.5 |
Mean |
0.043 |
0.019 |
-0.004 |
0.009 |
||
SD |
0.061 |
0.286 |
0.034 |
0.069 |
||||
N |
2 |
2 |
2 |
2 |
||||
G3 |
125 |
Mean |
0.014 |
0.057 |
-0.026 |
0.000 |
||
SD |
./. |
./. |
./. |
./. |
||||
N |
1 |
1 |
1 |
1 |
||||
G4 |
250 |
Mean |
0.129 |
0.024 |
-0.029 |
0.024 |
||
SD |
0.045 |
0.160 |
0.153 |
0.069 |
||||
N |
4 |
4 |
4 |
4 |
Key: SD: Standard Deviation; N: Number of Animals; ./.: not applicable
Table 2.7 Detailed clinical examination
|
|
Sex: Male |
|
|||||||||||||||||||
Group |
Dose (mg/Kg) |
Day of Exposure |
|
|||||||||||||||||||
1 |
8 |
15 |
22 |
29 |
36 |
43 |
50 |
57 |
64 |
71 |
|
|||||||||||
G1 |
0 |
NAD |
NAD |
NAD |
NAD |
NAD |
NAD |
NAD |
NAD |
NAD |
NAD |
NAD |
|
|||||||||
G2 |
62.5 |
NAD |
NAD |
NAD |
NAD |
NAD |
NAD |
NAD |
NAD |
NAD |
NAD |
NAD |
|
|||||||||
G3 |
125 |
NAD |
NAD |
NAD |
NAD |
NAD |
NAD |
NAD |
NAD |
NAD |
NAD |
NAD |
|
|||||||||
G4 |
250 |
NAD |
NAD |
NAD |
NAD |
NAD |
NAD |
NAD |
NAD |
NAD |
NAD |
NAD |
|
|||||||||
|
Pre-mating |
Sex: Female |
||||||||||||||||||||
Group |
Dose (mg/Kg) |
Day of Exposure |
||||||||||||||||||||
1 |
8 |
15 |
22 |
|||||||||||||||||||
G1 |
0 |
NAD |
NAD |
NAD |
NAD |
|||||||||||||||||
G2 |
62.5 |
NAD |
NAD |
NAD |
NAD |
|||||||||||||||||
G3 |
125 |
NAD |
NAD |
NAD |
NAD |
|||||||||||||||||
G4 |
250 |
NAD |
NAD |
NAD |
NAD |
|
Gestation |
Sex: Female |
|||||||||||
Group |
Dos (mg/Kg) |
Day of Gestation |
|||||||||||
0 |
7 |
14 |
20 |
||||||||||
G1 |
0 |
NAD |
NAD |
NAD |
NAD |
||||||||
G2 |
62.5 |
NAD |
NAD |
NAD |
NAD |
||||||||
G3 |
125 |
NAD |
NAD |
NAD |
NAD |
||||||||
G4 |
250 |
NAD |
NAD |
NAD |
NAD |
Key: NAD: no abnormality detected
|
Lactation |
Sex: Female |
||||||
Group |
Dose (mg/Kg) |
Day of Lactation |
||||||
0 |
4 |
7 |
14 |
21 |
||||
G1 |
0 |
NAD |
NAD |
NAD |
NAD |
NAD |
||
G2 |
62.5 |
NAD |
NAD |
NAD |
NAD |
NAD |
||
G3 |
125 |
NAD |
NAD |
NAD |
NAD |
NAD |
||
G4 |
250 |
NAD |
NAD |
NAD |
NAD |
NAD |
|
Non-Parturated |
Sex: Female |
||||||||
Group |
Dose (mg/Kg) |
Day of Gestation |
||||||||
0 |
7 |
14 |
20 |
27 |
30 |
34 |
||||
G1 |
0 |
NAD |
NAD |
NAD |
NAD |
NAD |
NAD |
NAD |
||
G2 |
62.5 |
NAD |
NAD |
NAD |
NAD |
NAD |
NAD |
NAD |
||
G3 |
125 |
NAD |
NAD |
NAD |
NAD |
NAD |
NAD |
NAD |
||
G4 |
250 |
NAD |
NAD |
NAD |
NAD |
NAD |
NAD |
NAD |
Table 2.8 Mean Hormone levels (T4, TSH, Estradiol, Progesterone, Testesterone)
|
|
Sex: Male |
|||||||
Group |
Dose (mg/kg) |
|
TSH(UI/ml) |
Estradiol (pg/ml) |
Progesterone (ng/ml) |
Tetsesterone (pg/ml) |
T4 (ng/ml) |
||
G1 |
0 |
Mean |
0.17 |
114.02 |
3.74 |
37.18 |
81.36 |
||
SD |
0.06 |
24.85 |
0.86 |
10.58 |
18.84 |
||||
N |
10 |
10 |
10 |
10 |
10 |
||||
G2 |
62.5 |
Mean |
0.15 |
100.65 |
3.46 |
39.36 |
64.34 |
||
SD |
0.03 |
24.63 |
0.50 |
15.81 |
13.55 |
||||
N |
10 |
10 |
10 |
10 |
10 |
||||
G3 |
125 |
Mean |
0.14 |
97.43 |
3.47 |
32.89 |
53.49↓↓ |
||
SD |
0.07 |
27.86 |
1.12 |
12.84 |
11.01 |
||||
N |
10 |
10 |
10 |
10 |
10 |
||||
G4 |
250 |
Mean |
0.12 |
104.89 |
3.97 |
32.22 |
69.79 |
||
SD |
0.03 |
19.89 |
1.03 |
16.61 |
21.42 |
||||
N |
10 |
10 |
10 |
9 |
10 |
|
|
Sex: Female |
|||||||
Group |
Dose (mg/kg) |
|
TSH(UI/ml) |
Estradiol (pg/ml) |
Progesterone (ng/ml) |
Tetsesterone (pg/ml) |
T4 (ng/ml) |
||
G1 |
0 |
Mean |
0.12 |
104.81 |
6.26 |
52.54 |
49.27 |
||
SD |
0.02 |
27.48 |
1.34 |
10.91 |
8.33 |
||||
N |
10 |
10 |
10 |
10 |
10 |
||||
G2 |
62.5 |
Mean |
0.10 |
97.80 |
5.76 |
65.59 |
51.60 |
||
SD |
0.03 |
24.62 |
2.07 |
16.44 |
6.71 |
||||
N |
10 |
10 |
10 |
10 |
10 |
||||
G3 |
125 |
Mean |
0.12 |
103.23 |
5.10 |
54.71 |
56.61 |
||
SD |
0.03 |
22.84 |
1.56 |
16.05 |
8.09 |
||||
N |
10 |
10 |
10 |
10 |
10 |
||||
G4 |
250 |
Mean |
0.10 |
84.89 |
3.99↓↓↓ |
45.70 |
47.63 |
||
SD |
0.02 |
25.31 |
1.56 |
9.51 |
11.29 |
||||
N |
10 |
10 |
10 |
10 |
10 |
Key: N: number of animals, SD: standard deviation; ↓↓: decreased as compared to control (P<0.01); ↓↓↓: decreased as compared to control (P<0.001).
Table 2.9 Hematology
Sex: Male |
||||||||||
Group (N) |
G1 (10) |
G2 (10) |
G3 (10) |
G4 (10) |
||||||
Dose (mg/kg) |
0 |
62.5 |
125 |
250 |
||||||
Parameters |
Mean |
SD |
Mean |
SD |
Mean |
SD |
Mean |
SD |
|
|
Total Erythrocyte Count (RBC) (X 1012/L) |
8.62 |
0.49 |
8.82 |
0.41 |
8.47 |
0.39 |
8.76 |
0.48 |
|
|
Hematocrit (HCT) (%) |
43.61 |
1.61 |
44.22 |
2.05 |
42.28 |
2.43 |
43.41 |
1.85 |
|
|
Mean Corpuscular Volume (MCV) (fL) |
50.69 |
2.78 |
50.11 |
1.37 |
49.90 |
1.39 |
49.57 |
1.97 |
|
|
Hemoglobin (HGB) (g/dL) |
15.62 |
0.69 |
15.84 |
0.69 |
15.19 |
0.90 |
15.56 |
0.60 |
|
|
Mean Corpuscular Hemoglobin (MCH) (pg) |
18.16 |
0.91 |
17.95 |
0.40 |
17.92 |
0.47 |
17.77 |
0.68 |
|
|
Mean Corpuscular Hemoglobin Concentration (MCHC) (g/dL) |
35.84 |
0.45 |
35.88 |
0.39 |
35.93 |
0.25 |
35.88 |
0.66 |
|
|
Platelet Count (PLT) (X 109/L) |
676.30 |
63.08 |
682.20 |
86.96 |
686.90 |
71.09 |
705.80 |
49.65 |
|
|
Total Leukocyte count (WBC) (X 109/L) |
9.33 |
0.99 |
10.29 |
1.84 |
10.27 |
3.32 |
12.50↑↑ |
1.93 |
|
|
Neutrophil (%) |
18.20 |
2.35 |
18.50 |
3.24 |
17.70 |
2.58 |
18.00 |
2.67 |
|
|
Lymphocyte (%) |
81.30 |
2.26 |
81.00 |
3.16 |
82.00 |
2.40 |
81.10 |
2.42 |
|
|
Monocyte (%) |
0.20 |
0.42 |
0.30 |
0.48 |
0.30 |
0.48 |
0.60 |
0.52 |
|
|
Eosinophil (%) |
0.30 |
0.48 |
0.20 |
0.42 |
0.00 |
0.00 |
0.30 |
0.48 |
|
|
Basophil (%) |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
|
|
Prothrombin Time (PT) (Sec.) |
22.22 |
2.43 |
23.31 |
1.93 |
22.98 |
2.01 |
22.00 |
1.11 |
|
|
Activated Partial thromboplastin Time (aPTT) (Sec.) |
28.45 |
1.76 |
27.44 |
1.12 |
28.33 |
1.58 |
27.07 |
1.72 |
|
|
Reticulocytes (%) |
1.00 |
0.07 |
1.00 |
0.05 |
1.05 |
0.10 |
1.00 |
0.05 |
|
Keys: SD = Standard Deviation; N = Number of animals; ↑↑: increased as compared to control (P<0.01).
Sex: Female |
||||||||||
Group (N) |
G1 (10) |
G2 (10) |
G3 (10) |
G4 (10) |
||||||
Dose (mg/kg) |
0 |
62.5 |
125 |
250 |
||||||
Parameters |
Mean |
SD |
Mean |
SD |
Mean |
SD |
Mean |
SD |
|
|
Total Erythrocyte Count (RBC) (X 1012/L) |
7.91 |
0.52 |
7.77 |
0.50 |
7.99 |
0.36 |
7.85 |
0.22 |
|
|
Hematocrit (HCT) (%) |
42.17 |
2.70 |
41.33 |
2.51 |
42.93 |
1.55 |
42.41 |
1.61 |
|
|
Mean Corpuscular Volume (MCV) (fL) |
53.34 |
2.41 |
53.25 |
1.78 |
53.71 |
2.08 |
54.01 |
1.54 |
|
|
Hemoglobin (HGB) (g/dL) |
14.97 |
0.91 |
14.79 |
0.85 |
15.29 |
0.43 |
15.04 |
0.55 |
|
|
Mean Corpuscular Hemoglobin (MCH) (pg) |
18.95 |
0.66 |
19.06 |
0.49 |
19.14 |
0.61 |
19.16 |
0.61 |
|
|
Mean Corpuscular Hemoglobin Concentration (MCHC) (g/dL) |
35.52 |
0.71 |
35.84 |
0.55 |
35.67 |
0.62 |
35.46 |
0.27 |
|
|
Platelet Count (PLT) (X 109/L) |
718.00 |
82.00 |
739.20 |
82.18 |
718.70 |
81.04 |
719.50 |
58.33 |
|
|
Total Leukocyte count (WBC) (X 109/L) |
9.36 |
3.28 |
11.50 |
5.01 |
12.88 |
3.70 |
13.14 |
3.52 |
|
|
Neutrophil (%) |
18.30 |
3.30 |
17.60 |
2.84 |
19.40 |
1.78 |
16.40 |
2.01 |
|
|
Lymphocyte (%) |
81.30 |
2.98 |
81.80 |
2.62 |
80.00 |
1.56 |
83.00 |
1.83 |
|
|
Monocyte (%) |
0.20 |
0.42 |
0.30 |
0.48 |
0.40 |
0.52 |
0.40 |
0.52 |
|
|
Eosinophil (%) |
0.20 |
0.42 |
0.30 |
0.48 |
0.20 |
0.42 |
0.20 |
0.42 |
|
|
Basophil (%) |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
|
|
Prothrombin Time (PT) (Sec.) |
20.83 |
1.70 |
21.08 |
1.56 |
21.69 |
1.53 |
22.45 |
2.89 |
|
|
Activated Partial thromboplastin Time (aPTT) (Sec.) |
28.39 |
1.53 |
27.18 |
1.24 |
27.17 |
1.37 |
28.25 |
1.42 |
|
|
Reticulocytes (%) |
1.01 |
0.10 |
1.03 |
0.12 |
1.08 |
0.14 |
1.08 |
0.12 |
|
Table 3.1 Clinical Chemistry
Sex: Male |
||||||||||
Group (N) |
G1 (10) |
G2 (10) |
G3 (10) |
G4 (10) |
||||||
Dose (mg/kg) |
0 |
62.5 |
125 |
250 |
||||||
Parameters |
Mean |
SD |
Mean |
SD |
Mean |
SD |
Mean |
SD |
|
|
ALT (U/l) |
49.95 |
6.63 |
44.29 |
5.47 |
46.29 |
7.59 |
55.30 |
5.71 |
|
|
AST (U/l) |
176.42 |
22.54 |
171.89 |
35.76 |
178.80 |
34.14 |
192.26 |
32.39 |
|
|
Albumin (g/dl) |
3.85 |
0.14 |
4.00 |
0.17 |
4.00 |
0.20 |
4.07 |
0.36 |
|
|
ALP (U/l) |
171.20 |
15.92 |
150.88 |
11.15 |
201.88 |
31.82 |
176.10 |
21.19 |
|
|
T.Protein (g/dl) |
6.32 |
0.63 |
6.54 |
0.59 |
6.45 |
0.47 |
6.67 |
0.52 |
|
|
Urea (mg/dl) |
49.76 |
7.90 |
49.48 |
3.88 |
48.13 |
12.59 |
51.20 |
10.64 |
|
|
Bile Acid (µmol/l) |
21.62 |
11.11 |
25.12 |
10.76 |
27.38 |
9.23 |
23.67 |
11.07 |
|
|
Cholesterol (mg/dl) |
53.21 |
7.92 |
55.99 |
7.83 |
60.08 |
11.93 |
58.38 |
8.37 |
|
|
Calcium (mg/dl) |
10.11 |
0.31 |
10.12 |
0.60 |
10.30 |
0.25 |
10.37 |
0.51 |
|
|
CK- Nac (U/I) |
780.10 |
183.19 |
813.90 |
209.05 |
803.40 |
228.85 |
733.80 |
177.29 |
|
|
Creatinine (mg/dl) |
0.53 |
0.05 |
0.53 |
0.02 |
0.51 |
0.04 |
0.51 |
0.04 |
|
|
Glucose (mg/dl) |
88.59 |
11.28 |
84.55 |
9.86 |
88.30 |
5.54 |
92.58 |
12.36 |
|
|
GGT(U/I) |
4.93 |
1.81 |
5.60 |
2.33 |
5.12 |
2.85 |
5.14 |
2.60 |
|
|
LDH(U/I) |
1131.39 |
225.25 |
1042.27 |
286.03 |
1028.03 |
238.64 |
1146.85 |
248.37 |
|
|
Phosphorus(mg/dl) |
6.06 |
0.83 |
5.62 |
0.74 |
6.15 |
0.65 |
7.04↑ |
1.08 |
|
|
Triglyceride (mg/dl) |
65.59 |
15.66 |
65.37 |
10.46 |
60.29 |
6.70 |
60.00 |
9.70 |
|
|
Total Bilirubin (mg/dl) |
0.33 |
0.16 |
0.25 |
0.08 |
0.29 |
0.11 |
0.32 |
0.05 |
|
|
BUN (mg/dl) |
23.25 |
3.69 |
23.10 |
1.84 |
22.50 |
5.88 |
23.45 |
4.92 |
|
|
A/G Ratio |
1.65 |
0.39 |
1.67 |
0.50 |
1.68 |
0.30 |
1.63 |
0.42 |
|
|
Globulin (g/dl) |
2.46 |
0.57 |
2.55 |
0.61 |
2.45 |
0.41 |
2.60 |
0.50 |
|
|
Na (mmol/l) |
145.00 |
2.05 |
142.40 |
4.84 |
147.20 |
2.44 |
149.60↑ |
4.95 |
|
|
K (mmol/l) |
5.87 |
0.79 |
5.99 |
0.93 |
5.85 |
0.52 |
5.76 |
0.44 |
|
|
Cl (mmol/l) |
99.30 |
2.36 |
99.10 |
4.33 |
101.70 |
2.45 |
104.00↑ |
3.94 |
|
Sex: Female |
||||||||||
Group (N) |
G1 (10) |
G2 (10) |
G3 (10) |
G4 (10) |
||||||
Dose (mg/kg) |
0 |
62.5 |
125 |
250 |
||||||
Parameters |
Mean |
SD |
Mean |
SD |
Mean |
SD |
Mean |
SD |
|
|
ALT (U/l) |
80.68 |
11.99 |
87.19 |
18.28 |
79.58 |
13.05 |
88.76 |
17.95 |
|
|
AST (U/l) |
184.60 |
23.52 |
192.07 |
21.92 |
186.66 |
23.20 |
192.09 |
20.06 |
|
|
Albumin (g/dl) |
3.94 |
0.35 |
3.78 |
0.31 |
3.88 |
0.28 |
3.78 |
0.33 |
|
|
ALP (U/l) |
148.25 |
29.22 |
132.24 |
17.25 |
136.98 |
20.93 |
144.91 |
21.30 |
|
|
T.Protein (g/dl) |
7.15 |
0.66 |
7.02 |
0.52 |
6.92 |
0.67 |
6.95 |
0.75 |
|
|
Urea (mg/dl) |
70.14 |
7.13 |
65.41 |
10.72 |
64.40 |
7.58 |
65.18 |
12.35 |
|
|
Bile Acid (µmol/l) |
55.72 |
13.92 |
52.76 |
19.48 |
50.90 |
20.35 |
50.62 |
17.47 |
|
|
Cholesterol (mg/dl) |
74.43 |
10.30 |
78.96 |
10.13 |
80.41 |
13.27 |
77.15 |
12.86 |
|
|
Calcium (mg/dl) |
9.52 |
0.30 |
9.68 |
0.51 |
10.02 |
0.41 |
9.58 |
0.51 |
|
|
CK- Nac (U/I) |
647.00 |
216.82 |
660.10 |
183.83 |
691.60 |
180.56 |
689.30 |
186.26 |
|
|
Creatinine (mg/dl) |
1.29 |
0.26 |
1.37 |
0.25 |
1.31 |
0.25 |
1.23 |
0.26 |
|
|
Glucose (mg/dl) |
89.82 |
8.90 |
96.87 |
11.33 |
95.40 |
12.38 |
91.79 |
13.60 |
|
|
GGT(U/I) |
9.49 |
2.31 |
8.08 |
2.90 |
8.43 |
1.69 |
8.01 |
1.51 |
|
|
LDH(U/I) |
1083.00 |
232.85 |
988.32 |
258.29 |
1108.26 |
244.02 |
1120.21 |
279.06 |
|
|
Phosphorus(mg/dl) |
8.01 |
1.03 |
7.70 |
1.08 |
7.87 |
1.35 |
7.88 |
0.58 |
|
|
Triglyceride (mg/dl) |
61.52 |
19.01 |
65.80 |
12.71 |
63.31 |
9.60 |
67.74 |
10.02 |
|
|
Total Bilirubin (mg/dl) |
0.50 |
0.09 |
0.64 |
0.16 |
0.64 |
0.26 |
0.56 |
0.17 |
|
|
BUN (mg/dl) |
32.77 |
3.34 |
30.58 |
5.01 |
30.00 |
3.49 |
30.46 |
5.77 |
|
|
A/G Ratio |
1.20 |
0.16 |
1.19 |
0.18 |
1.29 |
0.15 |
1.23 |
0.21 |
|
|
Globulin (g/dl) |
3.28 |
0.56 |
3.23 |
0.43 |
3.04 |
0.45 |
3.17 |
0.62 |
|
|
Na (mmol/l) |
139.80 |
1.48 |
139.60 |
0.70 |
139.30 |
2.21 |
138.70 |
2.00 |
|
|
K (mmol/l) |
6.54 |
0.77 |
6.21 |
0.73 |
6.57 |
0.79 |
6.29 |
0.69 |
|
|
Cl (mmol/l) |
99.00 |
2.21 |
99.50 |
2.46 |
100.60 |
2.01 |
100.30 |
2.58 |
|
Table 3.2 Urinalysis
Sex: Male |
||||||||||
Group (N) |
G1 (10) |
G2 (10) |
G3 (10) |
G4 (10) |
||||||
Dose (mg/kg) |
0 |
62.5 |
125 |
250 |
||||||
Parameters |
Mean |
SD |
Mean |
SD |
Mean |
SD |
Mean |
SD |
|
|
Blood / Blood Cell (RBC/µL) |
neg |
./. |
neg |
./. |
neg |
./. |
neg |
./. |
|
|
Bilirubin (mg/dL) |
neg |
./. |
neg |
./. |
neg |
./. |
neg |
./. |
|
|
Urobilinogen (mg/dL) |
norm |
./. |
norm |
./. |
norm |
./. |
norm |
./. |
|
|
Ketone (mg/dL) |
neg |
./. |
neg |
./. |
neg |
./. |
neg |
./. |
|
|
Protein (mg/dL) |
neg |
./. |
neg |
./. |
neg |
./. |
neg |
./. |
|
|
Nitrite |
neg |
./. |
neg |
./. |
neg |
./. |
neg |
./. |
|
|
Glucose (mg/dL) |
neg |
./. |
neg |
./. |
neg |
./. |
neg |
./. |
|
|
pH |
5.90 |
0.74 |
5.85 |
0.47 |
5.80 |
0.48 |
6.00 |
0.62 |
|
|
Specific Gravity |
1.022 |
0.005 |
1.017 |
0.006 |
1.020 |
0.006 |
1.019 |
0.007 |
|
|
Leukocytes (WBC/µL) |
neg |
./. |
neg |
./. |
neg |
./. |
neg |
./. |
|
|
Physical Observations |
||||||||||
Volume (mL) |
16.40 |
3.86 |
16.80 |
3.05 |
15.20 |
2.90 |
16.80 |
3.79 |
|
|
Colour |
Pale Yellow |
./. |
Pale Yellow |
./. |
Pale Yellow |
./. |
Pale Yellow |
./. |
|
|
Appearance |
Clear |
./. |
Clear |
./. |
Clear |
./. |
Clear |
./. |
|
Keys: neg:negative; norm: normal
Sex: Female |
||||||||||
Group (N) |
G1 (10) |
G2 (10) |
G3 (10) |
G4 (10) |
||||||
Dose (mg/kg) |
0 |
62.5 |
125 |
250 |
||||||
Parameters |
Mean |
SD |
Mean |
SD |
Mean |
SD |
Mean |
SD |
|
|
Blood / Blood Cell (RBC/µL) |
neg |
./. |
neg |
./. |
neg |
./. |
neg |
./. |
|
|
Bilirubin (mg/dL) |
neg |
./. |
neg |
./. |
neg |
./. |
neg |
./. |
|
|
Urobilinogen (mg/dL) |
norm |
./. |
norm |
./. |
norm |
./. |
norm |
./. |
|
|
Ketone (mg/dL) |
neg |
./. |
neg |
./. |
neg |
./. |
neg |
./. |
|
|
Protein (mg/dL) |
neg |
./. |
neg |
./. |
neg |
./. |
neg |
./. |
|
|
Nitrite |
neg |
./. |
neg |
./. |
neg |
./. |
neg |
./. |
|
|
Glucose (mg/dL) |
neg |
./. |
neg |
./. |
neg |
./. |
neg |
./. |
|
|
pH |
5.35 |
0.47 |
5.30 |
0.42 |
5.45 |
0.50 |
5.25 |
0.42 |
|
|
Specific Gravity |
1.024 |
0.002 |
1.025 |
0.000 |
1.024 |
0.002 |
1.024 |
0.002 |
|
|
Leukocytes (WBC/µL) |
neg |
./. |
neg |
./. |
neg |
./. |
neg |
./. |
|
|
Physical Observations |
||||||||||
Volume (mL) |
14.10 |
3.60 |
14.80 |
2.25 |
15.60 |
1.71 |
16.30 |
3.16 |
|
|
Colour |
Pale Yellow |
./. |
Pale Yellow |
./. |
Pale Yellow |
./. |
Pale Yellow |
./. |
|
|
Appearance |
Clear |
./. |
Clear |
./. |
Clear |
./. |
Clear |
./. |
|
Keys: neg: negative; norm: normal
Table 3.3 Absolute organ weights
Mean organ weights (g) |
Sex: Male |
||||||||||||||||
Group |
Dose |
BW |
Brain |
Adrenals |
P & SV |
Testes |
Ep |
Heart |
Liver |
Kidneys |
Spleen |
Thymus |
Pituitary |
T & PT |
|||
G1 |
0 |
Mean |
432.13 |
2.1733 |
0.0653 |
3.8409 |
3.2878 |
1.5106 |
1.3489 |
13.2371 |
2.8793 |
0.8518 |
0.3846 |
0.0148 |
0.0225 |
||
SD |
26.85 |
0.0938 |
0.0115 |
0.4526 |
0.2009 |
0.1184 |
0.1111 |
1.4264 |
0.4488 |
0.4390 |
0.0820 |
0.0021 |
0.0031 |
||||
N |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
||||
G2 |
62.5 |
Mean |
439.10 |
2.1894 |
0.0659 |
3.7032 |
3.3069 |
1.5295 |
1.3957 |
13.6968 |
2.9693 |
0.7618 |
0.4097 |
0.0146 |
0.0219 |
||
SD |
27.80 |
0.1205 |
0.0104 |
0.3724 |
0.2647 |
0.1288 |
0.1500 |
1.7457 |
0.1780 |
0.0994 |
0.0887 |
0.0017 |
0.0041 |
||||
N |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
||||
G3 |
125 |
Mean |
432.57 |
2.2122 |
0.0688 |
3.5891 |
3.4090 |
1.5136 |
1.3780 |
13.9272 |
3.0300 |
0.8931 |
0.4029 |
0.0145 |
0.0224 |
||
SD |
32.71 |
0.0822 |
0.0088 |
0.5126 |
0.2741 |
0.1457 |
0.1242 |
1.3925 |
0.2507 |
0.4175 |
0.0688 |
0.0020 |
0.0038 |
||||
N |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
||||
G4 |
250 |
Mean |
417.10 |
2.1821 |
0.0653 |
3.5503 |
3.2489 |
1.4888 |
1.3824 |
13.3019 |
2.9758 |
0.7588 |
0.3749 |
0.0138 |
0.0220 |
||
SD |
36.89 |
0.0900 |
0.0101 |
0.4892 |
0.4676 |
0.1595 |
0.1867 |
1.7709 |
0.3089 |
0.0928 |
0.0990 |
0.0016 |
0.0048 |
||||
N |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
Mean organ weights (g) |
Sex: Female |
|||||||||||||||
Group |
Dose |
BW |
Brain |
Adrenals |
Ovaries |
Uterus |
Heart |
Liver |
Kidneys |
Spleen |
Thymus |
Pituitary |
T & PT |
|||
G1 |
0 |
Mean |
277.13 |
2.0061 |
0.0883 |
0.1506 |
0.4639 |
1.0677 |
15.8516 |
2.1307 |
0.6678 |
0.2184 |
0.0137 |
0.0191 |
||
SD |
22.36 |
0.0991 |
0.0131 |
0.0368 |
0.1418 |
0.1691 |
3.0913 |
0.2136 |
0.1410 |
0.1030 |
0.0021 |
0.0039 |
||||
N |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
||||
G2 |
62.5 |
Mean |
280.60 |
2.0174 |
0.0861 |
0.1476 |
0.4310 |
1.0576 |
16.0515 |
2.0773 |
0.6261 |
0.2262 |
0.0139 |
0.0179 |
||
SD |
22.07 |
0.1135 |
0.0114 |
0.0252 |
0.1315 |
0.0815 |
3.4609 |
0.1922 |
0.0792 |
0.0813 |
0.0019 |
0.0042 |
||||
N |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
||||
G3 |
125 |
Mean |
278.37 |
1.9954 |
0.0873 |
0.1484 |
0.4800 |
1.0546 |
16.2290 |
2.0188 |
0.7056 |
0.2027 |
0.0137 |
0.0187 |
||
SD |
24.92 |
0.1270 |
0.0106 |
0.0308 |
0.1354 |
0.1175 |
3.4161 |
0.3792 |
0.4207 |
0.0728 |
0.0020 |
0.0036 |
||||
N |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
||||
G4 |
250 |
Mean |
271.83 |
1.9932 |
0.0875 |
0.1493 |
0.4865 |
1.0543 |
13.7392↓ |
2.0776 |
0.6942 |
0.2307 |
0.0146 |
0.0194 |
||
SD |
21.92 |
0.0826 |
0.0117 |
0.0227 |
0.1652 |
0.1286 |
2.8704 |
0.3441 |
0.2913 |
0.0681 |
0.0023 |
0.0032 |
||||
N |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
Key: BW = Body Weight; Ep = Epididymis; N = Number of samples; P & SV = Prostate and Seminal Vesicle; SD = Standard deviation; T & PT = Thyroid & Parathyroid. Dose levels are in terms of mg/kg body weight; ↓: decreased as compared to control (P<0.05).
Table 3.4 Relative organ weights
Mean organ weight relative body weight (%) |
Sex: Male |
|||||||||||||||
Group |
Dose |
Brain |
Adrenals |
P & SV |
Testes |
Ep |
Heart |
Liver |
Kidneys |
Spleen |
Thymus |
Pituitary |
T & PT |
|||
G1 |
0 |
Mean |
0.5046 |
0.0152 |
0.8920 |
0.7624 |
0.3504 |
0.3126 |
3.0619 |
0.6669 |
0.1975 |
0.0892 |
0.0034 |
0.0052 |
||
SD |
0.0354 |
0.0028 |
0.1175 |
0.0478 |
0.0291 |
0.0239 |
0.2536 |
0.1008 |
0.1037 |
0.0192 |
0.0004 |
0.0008 |
||||
N |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
||||
G2 |
62.5 |
Mean |
0.4999 |
0.0151 |
0.8449 |
0.7541 |
0.3487 |
0.3178 |
3.1250 |
0.6776 |
0.1740 |
0.0932 |
0.0033 |
0.0050 |
||
SD |
0.0321 |
0.0027 |
0.0845 |
0.0549 |
0.0255 |
0.0270 |
0.3947 |
0.0416 |
0.0239 |
0.0191 |
0.0004 |
0.0010 |
||||
N |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
||||
G3 |
125 |
Mean |
0.5141 |
0.0160 |
0.8327 |
0.7901 |
0.3509 |
0.3190 |
3.2271 |
0.7014 |
0.2081 |
0.0935 |
0.0034 |
0.0052 |
||
SD |
0.0411 |
0.0022 |
0.1216 |
0.0609 |
0.0336 |
0.0226 |
0.3076 |
0.0434 |
0.1036 |
0.0167 |
0.0005 |
0.0010 |
||||
N |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
||||
G4 |
250 |
Mean |
0.5266 |
0.0157 |
0.8536 |
0.7834 |
0.3589 |
0.3318 |
3.1888 |
0.7142 |
0.1831 |
0.0903 |
0.0033 |
0.0053 |
||
SD |
0.0451 |
0.0023 |
0.1126 |
0.1228 |
0.0442 |
0.0393 |
0.3091 |
0.0526 |
0.0258 |
0.0245 |
0.0004 |
0.0013 |
||||
N |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
Mean organ weight relative body weight (%) |
Sex: Female |
||||||||||||||
Group |
Dose |
Brain |
Adrenals |
Ovaries |
Uterus |
Heart |
Liver |
Kidneys |
Spleen |
Thymus |
Pituitary |
T & PT |
|||
G1 |
0 |
Mean |
0.7279 |
0.0321 |
0.0545 |
0.1686 |
0.3855 |
5.7142 |
0.7716 |
0.2431 |
0.0796 |
0.0049 |
0.0069 |
||
SD |
0.0621 |
0.0055 |
0.0136 |
0.0532 |
0.0518 |
0.9615 |
0.0797 |
0.0590 |
0.0388 |
0.0007 |
0.0015 |
||||
N |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
||||
G2 |
62.5 |
Mean |
0.7224 |
0.0308 |
0.0525 |
0.1529 |
0.3777 |
5.7142 |
0.7414 |
0.2233 |
0.0809 |
0.0050 |
0.0064 |
||
SD |
0.0596 |
0.0040 |
0.0067 |
0.0424 |
0.0239 |
1.0950 |
0.0554 |
0.0246 |
0.0299 |
0.0008 |
0.0014 |
||||
N |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
||||
G3 |
125 |
Mean |
0.7208 |
0.0315 |
0.0535 |
0.1743 |
0.3793 |
5.8108 |
0.7278 |
0.2525 |
0.0731 |
0.0050 |
0.0067 |
||
SD |
0.0619 |
0.0037 |
0.0109 |
0.0534 |
0.0310 |
1.0410 |
0.1324 |
0.1423 |
0.0277 |
0.0009 |
0.0011 |
||||
N |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
||||
G4 |
250 |
Mean |
0.7371 |
0.0324 |
0.0552 |
0.1794 |
0.3877 |
5.0395↓ |
0.7652 |
0.2558 |
0.0848 |
0.0054 |
0.0072 |
||
SD |
0.0590 |
0.0053 |
0.0088 |
0.0600 |
0.0325 |
0.8677 |
0.1217 |
0.1115 |
0.0238 |
0.0008 |
0.0013 |
||||
N |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
30 |
Key: Ep = Epididymis; N = Number of samples; P & SV = Prostate and Seminal Vesicle; SD = Standard deviation; T & PT = Thyroid & Parathyroid. Dose levels are in terms of mg/kg body weight; ↓: decreased as compared to control (P<0.05).
Table 3.5 Gross Pathology
|
Sex: Male |
||||
Group |
G1 |
G2 |
G3 |
G4 |
|
Dose (mg/kg) |
0 |
62.5 |
125 |
250 |
|
Total No. of Animals Observed |
30 |
30 |
30 |
30 |
|
No. of animals with Abnormalities |
0 |
2 |
1 |
1 |
|
No. of animals with No Abnormalities |
30 |
28 |
29 |
29 |
|
Organ & Lesion |
|||||
Testes |
|
|
|
|
|
Reduced sizeTotal |
0 |
1 |
0 |
1 |
|
Mild |
0 |
0 |
0 |
1 |
|
Moderate |
0 |
1 |
0 |
0 |
|
Prostate |
|
|
|
|
|
Increased sizeTotal |
0 |
1 |
0 |
0 |
|
Minimal |
0 |
1 |
0 |
0 |
|
Reduced sizeTotal |
0 |
0 |
1 |
0 |
|
Moderate |
0 |
0 |
1 |
0 |
|
Sex: Female |
||||
Group |
G1 |
G2 |
G3 |
G4 |
|
Dose (mg/kg) |
0 |
62.5 |
125 |
250 |
|
Total No. of Animals Observed |
30 |
30 |
30 |
30 |
|
No. of animals with Abnormalities |
0 |
0 |
0 |
0 |
|
No. of animals with No Abnormalities |
30 |
30 |
30 |
30 |
Table 3.6 Individual Animal Microscopic Observations
|
Sex: Male |
||||
Group |
G1 |
G2 |
G3 |
G4 |
|
Dose (mg/kg) |
0 |
62.5 |
125 |
250 |
|
Total No. of Animals Observed |
30 |
30 |
30 |
30 |
|
No. of animals with Abnormalities |
11 |
X |
X |
5 |
|
No. of animals with No Abnormalities |
19 |
X |
X |
25 |
|
Organ & Lesion |
|||||
Liver |
|
||||
Lymphocyte infiltrationTotal |
2 |
X |
X |
1 |
|
Focal Minimal |
0 |
X |
X |
1 |
|
Focal Mild |
2 |
X |
X |
0 |
|
Kidneys |
|
||||
Tubular dilation Total |
3 |
X |
X |
0 |
|
Focal Mild |
2 |
X |
X |
0 |
|
Focal Moderate |
1 |
X |
X |
0 |
|
Lymphocyte infiltration Total |
1 |
X |
X |
0 |
|
Focal Mild |
1 |
X |
X |
0 |
|
Lungs |
|
||||
Lymphocyte infiltration Total |
3 |
X |
X |
2 |
|
Multifocal Mild |
2 |
X |
X |
1 |
|
Multifocal Moderate |
1 |
X |
X |
1 |
|
Aorta |
|
||||
Aneurysm Total |
1 |
X |
X |
1 |
|
Focal Mild |
1 |
X |
X |
0 |
|
Focal Moderate |
0 |
X |
X |
1 |
|
Colon |
|
||||
Lymphocyte infiltration Total |
1 |
X |
X |
0 |
|
Multifocal Mild |
1 |
X |
X |
0 |
|
Thymus |
|
||||
LymphophagocytosisTotal |
1 |
X |
X |
0 |
|
Multifocal Minimal |
1 |
X |
X |
0 |
|
Thyroid |
|
||||
Ultimobranchial cystTotal |
1 |
X |
X |
1 |
|
Focal Mild |
1 |
X |
X |
1 |
|
Adrenals |
|
||||
Cytoplasmic vacuolationTotal |
1 |
X |
X |
0 |
|
Diffuse Mild |
1 |
X |
X |
0 |
|
Accessory adrenocortical tissueTotal |
1 |
X |
X |
0 |
|
Unilateral |
1 |
X |
X |
0 |
|
Sex: Female |
||||
Group |
G1 |
G2 |
G3 |
G4 |
|
Dose (mg/kg) |
0 |
62.5 |
125 |
250 |
|
Total No. of Animals Observed |
30 |
30 |
30 |
30 |
|
No. of animals with Abnormalities |
12 |
X |
X |
8 |
|
No. of animals with No Abnormalities |
18 |
X |
X |
22 |
|
Organ & Lesion |
|||||
Liver |
|
||||
Lymphocyte infiltration Total |
0 |
X |
X |
1 |
|
Focal Minimal |
0 |
X |
X |
1 |
|
Kidneys |
|
||||
Tubular dilation Total |
1 |
X |
X |
0 |
|
Multifocal Mild |
1 |
X |
X |
0 |
|
Lymphocyte infiltration Total |
2 |
X |
X |
1 |
|
Focal Minimal |
0 |
X |
X |
1 |
|
Focal Mild |
1 |
X |
X |
0 |
|
Multifocal Mild |
1 |
X |
X |
0 |
|
Lungs |
|
||||
Lymphocyte infiltration Total |
1 |
X |
X |
2 |
|
Focal Mild |
0 |
X |
X |
1 |
|
Multifocal Mild |
1 |
X |
X |
1 |
|
Alveolar histiocytosis Total |
1 |
X |
X |
1 |
|
Focal Mild |
0 |
X |
X |
1 |
|
Multifocal Mild |
1 |
X |
X |
0 |
|
Aorta |
|
||||
Aneurysm Total |
1 |
X |
X |
0 |
|
Focal Mild |
1 |
X |
X |
0 |
|
Thymus |
|
||||
LymphophagocytosisTotal |
1 |
X |
X |
1 |
|
Multifocal Mild |
1 |
X |
X |
1 |
|
Atrophy Total |
1 |
X |
X |
0 |
|
Focal Mild |
1 |
X |
X |
0 |
|
Thyroid |
|
||||
Ultimobranchial cystTotal |
2 |
X |
X |
1 |
|
Focal Mild |
2 |
X |
X |
0 |
|
Focal Moderate |
0 |
X |
X |
1 |
|
Adrenals |
|
|
|
|
|
Accessory adrenocortical tissueTotal |
2 |
X |
X |
2 |
|
Unilateral |
2 |
X |
X |
2 |
Applicant's summary and conclusion
- Conclusions:
- Based on all the available data, it was observed that there were no effects observed on male and female animals up to 125 mg/kg bw (Mid Dose-G3) of the test chemical administration. However, at 250 mg/kg bw (High Dose-G4) dose group, clinical signs, effects on weights of liver and effects on hormonal levels was observed, due to test chemical administration. Therefore, it was concluded that the NOAEL and LOAEL of the test chemical was 125 mg/kg bw and 250 mg/kg bw, respectively.
- Executive summary:
An Extended One-Generation Reproductive Toxicity Study was performed according to OECD TG 443 to evaluate the general, reproductive and developmental toxicological endpoints associated with the repeated oral administration of graduated doses of the test chemical in Wistar rats. A total of 240 wistar rats (120 males and 120 females) were randomized into 4 experimental groups each containing 30 animals per sex per group. The groups, viz., G1, G2, G3 and G4 received 0, 62.5, 125 and 250 mg/kg body weight of test chemical respectively. Dose administration was through an oral gavage and the vehicle used in this study was corn oil. All animals of both sexes were dosed 2 weeks prior to mating. Dosing was continued in both sexes during the mating period. Females were treated throughout gestation and lactation up to the day of termination after weaning. Males were treated in the same manner until termination. During observations, it was seen that no treatment-related clinical signs or symptoms were observed in any of the parent animals up to 125 mg/kg body weight of the test chemical. However, mild salivation was observed in total 13 of 30 males of G4 group (250 mg/kg), starting from the 27th day of treatment and continuing till termination. All adult animals were found normal with respect to all the parameters examined during detailed clinical examinations through the course of the study. No treatment-related symptoms were found up to the dose level of 250 mg/kg body weight in either sex. No morbidities or mortality were recorded in adult animals of parent generation in any of the experimental groups throughout the duration of the experiment. Mean body weights remained comparable among all the groups for every time point (days 8, 15, 22, 29, 36, 43, 50, 57, 64 and 71) recorded during the 70 days exposure period for males. No significant difference was found for the mean body weights of females during the two weeks pre-mating period, during mating, during gestation or during lactation period. However, the mean percent change in body weight on all the instances (days 8, 15, 22, 29, 36, 43, 50, 57, 64 and 71) with respect to the first day of exposure was significantly reduced for parental males of G4 group (250 mg/kg) (P≤0.05 to P≤0.001) as compared to control animals, whereas mean body weight change for G2 (62.5 mg/kg) and G3 (125 mg/kg) males remained comparable to control animals. Among the females, mean percent change in body weights remained statistically comparable among all groups during pre-mating, mating, gestation and lactation periods. The mean feed consumed per animal per day significantly increased on day 8 of treatment for G2 (62.5 mg/kg) (P≤0.01) and G3 (125 mg/kg) (P≤0.05) males as compared to G1 (control) animals. However, it was found to be similar for all other time points across the groups. The mean feed consumption of treatment group (G2, G3 & G4) females remained statistically comparable to control group during pre-mating, mating, gestation and lactation period except for one instance, where mean feed consumption on day 8 of treatment was significantly reduced for G2 (62.5 mg/kg) (P≤0.01) as compared to control females. The differences found in mean feed consumption of male and female animals were neither dose-dependent nor consistent, hence it was deemed to be independent of test-item treatment. Feed efficiency for the G4 males had statistically reduced in the first two weeks of treatment, i.e. on days 8 and 15. It was also found to have increased significantly during the penultimate week of treatment, i.e. on day 63. For all other groups among the males, the feed efficiency was statistically comparable across the groups. Among the parental females, no statistical differences were observed for feed efficiency in any of the groups. No significant difference was observed in any of the measured hematology parameters in all the groups of both the sexes, except in the WBC count, which was found to be significantly increased in males of G4 group (P≤0.01) as compared to control males. Significant increase was observed in Phosphorus, Sodium and Chloride levels in G4 males (P≤0.05) as compared to control males. All the other clinical chemistry parameters in males and females were found comparable in control and treatment groups. The urine parameters tested did not show up any significant changes among the experimental groups in the parental generation of either sex. No significant difference was observed in absolute organ weight or organ weight relative to body weight for the entire set of measured organs in parent male and female animals. However, significant reduction was observed in absolute and relative liver weight in G4 females (P<0.05) as compared to control animals. In the current study, however, the decreased liver weights with respect to body weights in G4 females does not correlate with any histological observations made, making it inconsequential from the point of view of toxic effects of test-chemical. External examination of all male and female rats of control and all treated groups did not reveal any lesion of pathological significance. Internal examination of the rats revealed reduced testicular size in two animals (Male: G2: 1/30, G4:1/30), increased prostate size in one animal (Male: G2: 1/30) and reduced prostate size in one animal (Male: G3: 1/30). Remaining animals of parent generation did not reveal any abnormality of pathological significance. No treatment-related histopathological findings are reported in this study that could arise out of test-item administration. All observed tissues were normal at the level of histology. Minimal to moderate and focal lymphocytic infiltrates were observed in some slides of liver, kidney, lung, heart, colon, thymus and adrenals with mild to moderate cyst in thyroid gland, which were distributed randomly across the experimental groups and did not show any pattern of either dose-dependency or sex-based selectivity. While TSH, Estradiol, Progesterone and Testosterone remained statistically comparable among the groups of parental males, the mean level of T4 was found reduced in G3 males (P<0.01) as compared to control. This is likely an incidental result, given that G4 was statistically comparable to control. The mean T4, TSH, Estradiol and testosterone levels remained comparable among the groups of parental females; whereas, Progesterone was found to be significantly reduced in G4 females (P<0.001). This could be a test-chemical related effect. Based on all the available data, it was observed that there were no effects observed on male and female animals up to 125 mg/kg bw (Mid Dose-G3) of the test chemical administration. However, at 250 mg/kg bw (High Dose-G4) dose group, clinical signs, effects on weights of liver and effects on hormonal levels was observed, due to test chemical administration. Therefore, it was concluded that the NOAEL and LOAEL of the test chemical was 125 mg/kg bw and 250 mg/kg bw, respectively.
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