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EC number: 201-972-0 | CAS number: 90-17-5
- 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
Toxicity to reproduction
Administrative data
- Endpoint:
- reproductive toxicity, other
- Remarks:
- reproductive organ toxicity study in rats
- 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 study report
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 014
- Report date:
- 2014
Materials and methods
Test guideline
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- other: OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents)
- Principles of method if other than guideline:
- The Repeated Dose 28-day Oral Toxicity study was designed and conducted to determine the reproductive toxicity profile of the test chemical when administered daily for 28 days in the Sprague Dawley rats.
- GLP compliance:
- yes
- Limit test:
- no
Test material
- Reference substance name:
- 2,2,2-trichloro-1-phenylethyl acetate
- EC Number:
- 201-972-0
- EC Name:
- 2,2,2-trichloro-1-phenylethyl acetate
- Cas Number:
- 90-17-5
- Molecular formula:
- C10H9Cl3O2
- IUPAC Name:
- 2,2,2-trichloro-1-phenylethyl acetate
- Details on test material:
- - Name of the test chemical: 2,2,2-TRICHLORO-1-PHENYLETHYL ACETATE
- Molecular formula: C10H9Cl3O2
- Molecular weight: 267.538 g/mol
- Substance type: Organic
Constituent 1
Test animals
- Species:
- rat
- Strain:
- Sprague-Dawley
- Details on species / strain selection:
- 1) In order to meet the regulatory requirement for testing in a rodent species
2) Widely used in as a species of choice for pre-clinical toxicological studies.
3) This strain is widely used throughout the industry in the non-clinical laboratory studies.
4) This study is intended to provide information on the health hazards likely to arise from exposure to the test item. - Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: National Institute of Biosciences, Pune
- Age at study initiation: 6 to 8 weeks
- Weight at study initiation:
Males: 132.8 g to 165.2 g
Females: 129.4 g to 156.4 g
- Fasting period before study: No Data
- Housing: The rats were housed in polycarbonate cages with paddy husk as bedding.
- Diet (e.g. ad libitum): Rodent feed supplied by the Nutrivet Life Sciences, Pune was provided ad libitum from individual feeders on cage top.
- Water (e.g. ad libitum): Water was provided ad libitum from individual bottles attached to the cages. Water was from a local source and passed through the reverse osmosis membrane before use.
- Acclimation period: 7 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): Room temperature was maintained at 22 ± 3°C (actual range, 19.6 °C to 21.4 °C).
- Humidity (%): Room humidity was maintained at 30% to 70% (actual range, 54.3% to 60.6%).
- Air changes (per hr): At least 10 Air changes per hour
- Photoperiod (hrs dark / hrs light): An artificial light and dark cycle of 12 hours each was provided to the room.
Administration / exposure
- Route of administration:
- oral: gavage
- Vehicle:
- polyethylene glycol
- Details on exposure:
- PREPARATION OF DOSING SOLUTIONS: The test item was suspended in Polyethylene Glycol-400 for preparation of suspension(s). The suspension(s) of test item was made at volumes suitable for daily use for 28 days. The solution(s) were prepared at concentrations of 0, 25, 50 and 100 mg/ml such that dosage of 0 (vehicle), 250, 500 and 1000 mg/kg body weight respectively were administered. The concentration of the test item was varied so as to maintain the dose volume constant at or upto 10 ml/kg body weight.
DIET PREPARATION
- Rate of preparation of diet (frequency): 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): Polyethylene glycol
- Concentration in vehicle: 0 (vehicle), 250, 500 and 1000 mg/kg body weight
- Amount of vehicle (if gavage): 10 mL/Kg
- Lot/batch no. (if required): No Data
- Purity: No Data - Details on mating procedure:
- No data available
- Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- The test item prepared formulation analysis for concentration, homogeneity and stability were conducted at subcontracted Laboratory.
- Duration of treatment / exposure:
- 28 Days consecutively and 2 week recovery period.
- Frequency of treatment:
- Once Daily
- Details on study schedule:
- No data available
Doses / concentrationsopen allclose all
- Dose / conc.:
- 0 mg/kg bw/day (actual dose received)
- Remarks:
- Control Group (G1)
- Dose / conc.:
- 250 mg/kg bw/day (actual dose received)
- Remarks:
- Low Dose Group (G2)
- Dose / conc.:
- 500 mg/kg bw/day (actual dose received)
- Remarks:
- Mid Dose Group (G3)
- Dose / conc.:
- 1 000 mg/kg bw/day (actual dose received)
- Remarks:
- High Dose Group (G4)
- No. of animals per sex per dose:
- Control (0 mg/kg bw/day): 6 males and 6 females
Control Recovery (0 mg/kg bw/day): 6 males and 6 females
Low dose (250 mg/Kg/Day): 6 males and 6 females.
Mid Dose (500 mg/Kg/Day): 6 males and 6 females
High Dose (1000 mg/Kg/Day): 6 males and 6 females
High Dose Recovery(1000 mg/Kg/Day): 6 males and 6 females - Control animals:
- yes, concurrent vehicle
- Details on study design:
- - Dose selection rationale: The doses were selected based on the results of the Dose Range Finder study conducted.The study was conducted at the dose levels of 0 mg/kg, 50 mg/kg, 100 mg/kg, 250 mg/kg, 500 mg/kg and 1000 mg/kg body weight.
1) All the male and female animals from control and all the treated dose groups up to 1000 mg/kg survived throughout the dosing period of 14 days.
2) Male and female animals from control and all the treated dose groups exhibited normal body weight gain at the end of the dosing period of 14 days.
3) Daily clinical observations did not reveal any signs of toxicity in male and female animals from different dose groups during the dosing period of 14 days.
4) Gross pathological examination did not reveal any abnormality attributable to the treatment.
Based on these results, the 28 day study dose levels were finalized as 0 mg/kg, 250 mg/kg, 500 mg/kg and 1000 mg/kg body weight and animals were exposed to the treatment, every day, for a period of 28 days.
- Rationale for animal assignment (if not random): Animals were randomized on the basis of sex and body weights. A total of 48 animals (24 males + 24 females) were selected and randomly distributed into four groups with 6 animals/sex/group and 3/sex/cage.
- Rationale for selecting satellite groups: No data
- Post-exposure recovery period in satellite groups: No data
- Section schedule rationale (if not random): No Data - Positive control:
- Not included
Examinations
- Parental animals: Observations and examinations:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Animals were observed twice daily for mortality. Rats were examined once daily for clinical signs.
- Cage side observations checked in table [No.?] were included. Viability, clinical signs such as skin and fur changes, eye and mucous membrane changes, respiratory, circulatory and general changes
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Detailed clinical observations were performed once before the start of dose administration and once a week thereafter until scheduled sacrifice.
a) Home Cage Observations:
In home cage, rats were observed for behavior, alterations, vocalizations, respiration and palpebral closer
1) Behavior in Home cage:
The behavior of the rat was observed in home cage upon initial approach by the observer and description of behavior in home cage was recorded as:
1 = apparently sleeping.
2 = Awake, but immobile; apparently normal posture.
3 = Engaged in apparently normal movement such as rearing, drinking, or grooming.
4 = Immobile, with unusual posture or tonic convulsion (lying on side with legs extended, flattened body or arched back. (opisthotonos or emprosthonos).
5 = Unusual behaviour or muscular patterns (stereotyped behaviour such as head bobbing or weaving, circling, repetitive licking or grooming, bizarre behaviour such as self mutilation, writhing or retropulsion, unusual muscular patterns such as tremors, spasms or clonic convulsion).
2) Alterations Home cage:
The alterations of the rat were observed in home cage upon initial approach by the observer and description of alterations in home cage was recorded as:
1 = No alterations in behaviour or posture (Normal).
2 = Stereotyped behaviour pattern (head bobbing or weaving, circling, repetitive licking or grooming).
3 = Bizarre behaviours such as self-mutilation, writhing or retropulsion.
4 = Twitches or tremors in the limbs or repetitive movements of the mouth or jaws.
5 = Whole body tremors or spasms.
6 = Unusual posture (opisthotonos, emprosthotonos, tonic extension, head tilt, straub tail).
7 = Tonic-clonic seizure.
3) Vocalizations:
The occurrence of spontaneous or unprovoked vocalization was recorded as:
1= No vocalization/ Normal
2= Vocalization noted
If vocalization observed the actual number was recorded.
4) Respiration:
The observations for respiration were recorded as:
1 = Normal
2 = Abnormal
The type of abnormal respiration e.g. bradypnea, hyperpnea, dyspnea, rals was recorded in the clinical signs.
5) Palpebral closer:
The degree of closure of the eyelids was recorded as:
1 = Eyelids wide open (Normal).
2 = Eyelids slightly closed.
3 = Eyelids dropping, approximately half-closed.
4 = Eyelids completely shut.
b) Handling Observations:
After completing home cage observations, the rat was observed for reaction to removal, reaction to handling, urination, defecation, prominence of eye, lacrimation, salivation, piloerection, examination of mucous membrane, examination of skin / fur, examination of natural orifices and animal appearance.
1) Reaction to Removal:
The reaction of the rat removed from home cage was recorded as:
1 = Sits quietly and is easily removed.
2 = Vocalization without resistance to being picked up.
3 = Runs around cage with or without vocalization, or freezes or rears, following the investigator’s hand.
4 = Tail and throat rattles, may attack.
2) Reaction to handling:
The reaction of the rat to handling from home cage was recorded as:
1 = Quiet with no resistance.
2 = Vocalization without resistance.
3 = Tense or rigid.
4 = Squirming and twisting, may attempt to bite.
3) Urination:
The frequency of urination was recorded as:
0 = No urination during the observation period.
1 = Urine present; quantity is not excessive.
2 = the amount of urination is excessive.
4) Defecation:
The frequency of defecation was recorded as:
0 = No defecation during the observation period.
1 = Fecal boluses have normal consistency.
2 = Soft or liquid feces.
5) Prominence of Eye:
The eyes were examined for prominence of eye and observation was recorded as:
1 = Normal
2 = Exophthalmos
3 = Enophthalmos
6) Lacrimation:
The degree of lacrimation was recorded as:
1 = No excess lacrimation (normal).
2 = Excess moisture at the margin of the eyelid.
3 = Persistent dampness at the margin of the eyelid
4 = Dampness extends beyond the margin of the eyelid.
7) Salivation:
The degree of salivation was recorded as:
1 = No excess salivation (normal).
2 = Margin of mouth wet.
3 = Wet zone ¼ to ½ of submandibular area.
4 = Wet zone extends to the entire submandibular area.
8) Piloerection:
Piloerection was differentiated from a scruffy or ungroomed coat by patting the back of the animal in a rostral to caudal direction. Piloerection was considered in case the animal hairs were erect after patting. The observation for piloerection was recorded as:
0 = Absent
1 = Present
9) Examination of Mucous Membrane:
The observation for visual mucous membrane was recorded as:
¬¬¬1 = Normal
2 = Abnormal (discolouration)
10) Examination of Skin / Fur:
The observation for skin / fur examination was recorded as:
1 = Normal
2 = Abnormal
11) Examination of Natural Orifices:
The observation for natural orifices examination was recorded as:
1 = Normal
2 = Abnormal
12) Animal Appearance:
The observation for appearance of animal was recorded as:
1 = Clean and groomed.
2 = Unkempt (with scruffy and ungroomed coat)
3 = Stained by urine and/or feces.
c) Open field observation:
The animal was placed in the open field and its appearance and behavior were observed. The following observations were made and recorded:
1) Stereotype Behaviour:
The stereotype behaviour can be defined as the pronounced repetition of specific gesture or movements i.e. presence of excessive or repetitive behaviour that appears purposeless to the observer. The observation was recorded as:
0 = Absent
1 = Excessive grooming / licking / head bobbing or weaving
2 = Circling movements
2) Bizarre Behaviour:
The bizarre behaviour includes any unusual behaviour that will not be normally observed in the test species. The observation was recorded as:
0 = Absent
1 = Retropulsion
2 = Biting of cage
3 = Biting to other animal(s)
4 = Self destructive biting or mutilation
3) Rearing (Rears):
The number of times the rat raises both forelimbs off the surfaces is considered as rearing. The number of these actions was counted and total number of rearing was recorded.
4) Movements:
In the open field, each animal was observed for presence of clonic and tonic movements:
4.1) Clonic Movements:
The observation for clonic movement was recorded as:
0 = None /Normal
1 = repetitive mouth/jaw motion, such as, Chewing, clones of jaw
2 = Mild clonic tremors of whole body
3 = Repetitive clonic tremors/ seizure of whole body
4.2) Tonic Movements:
The observation for tonic movement was recorded as:
0 = None /Normal
1 = Contractions of limbs
2 =Unusual posture (Opisthotonos, Emprosthotonous, tonic extension, head tilt, straub tail).
3= seizure
5) Gait Pattern:
The gait pattern was evaluated by observing the movement of the rat in the open field and the observation was recorded as:
1 = Normal
2 = Ataxic
3 = Hind limbs or forelimbs show exaggerated or overcompensated movements, drag or appear splayed.
4 = Spastic
5 = Duck walk
6 = Scissor
7 = Hunched back
6) Mobility Score:
A measure of the ability of the animal to locomote despite gait abnormalities was recorded. The ranking of the degree of impairment of locomotion was recorded as:
1 = Normal
2 = Slightly impaired
3 = Totally impaired
7) Severity of Gait:
The severity of the gait abnormalities is graded and documented as follows:
1 = Slight gait abnormality
2 = Moderate gait abnormality
3 = Extreme gait abnormality
8) Pupillary response:
The animal eyes were briefly covered for 30 seconds with hand/cloth and then the penlight was pointed and the response to penlight was recorded as:
1 = Response
2 = No response
BODY WEIGHT: Yes
- Time schedule for examinations: Body weights were recorded on the day of randomization, day of first dosing, weekly thereafter and a fasting body weight at scheduled sacrifice on day 29 and day 43.
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes, The quantity of feed consumed by control and different treatment groups was recorded on commencement of treatment and weekly thereafter until scheduled sacrifice and the feed consumption per animal was calculated for each group.
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data
FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No data
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data
- Time schedule for examinations: No Data
OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: The eyes of all the animals were examined prior to the initiation of the dosing and at scheduled sacrifice.
- Dose groups that were examined: All animals were examined.
HAEMATOLOGY: Yes
- Time schedule for collection of blood: Blood samples were collected at terminations.
- Anaesthetic used for blood collection: No data
- Animals fasted: Yes, Food was withheld overnight from all rats prior to sampling.
- How many animals: Blood samples were collected from all rats from each group.
- Parameters checked in table [No.?] were examined. Hb, RBC, HCT, MCV, MCH, MCHC, WBC, N, L, E, M, B, Pt
CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Blood samples were collected at respective terminations.
- Animals fasted: Yes, Food was withheld overnight from all rats prior to sampling.
- How many animals: Blood samples were collected from all rats from each group.
- Parameters checked in table [No.?] were examined. Total Protein , Blood Urea Nitrogen , Urea Nitrogen, ALT, AST, ALP, GGT, Glucose, Ca, P, Albumin, Total bilirubin, creatinine, total cholesterol, triglycerides, globulin, Na, K, Cl
URINANALYSIS: No data
- Time schedule for collection of urine: No data
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data
- Parameters checked in table [No.?] were examined. No data
NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: Towards the end of the exposure period of 28 days and towards the end of the recovery period on day 42.
- Dose groups that were examined: All animals from all groups.
- Battery of functions tested:
Sensory activity: Towards the end of the exposure period, sensory reactivity to stimuli of different types (e.g., auditory, visual and proprioceptive stimuli) was conducted for all the animals.
1) Arousal level:
The activity/arousal level of the animal was described during the observation period and is documented as follows:
1 = Very low (stuporous, comatose)
2 = Low (sluggish, some exploratory movement possible).
3 = Apparently normal (alert with exploratory movement).
4 = High (sudden startle, darting or freezing without apparent stimuli).
5 = Very high (sudden bouts of running, freezing with spontaneous vocalization).
2) Sensory Activity:
Sensory activity was assessed by following methods:
2.1) Assessment of Visual Response: A blunt probe was held approximately 4 cm away from front of the face / eye and moved away steadily and reaction was documented.
2.2) Touch Response: Avoiding the animal’s field of vision, the rump was gently touched with blunt probe. The animals reaction to this stimulus was observed and was documented.
2.3) Auditory Response: Using a clicker approximately 5 cm above the back of the animal sudden sound was made. The animals reaction to this stimulus was observed and was documented.
2.4) Tail Pinch Response: The reaction to a tail pinch was rated (The tail pinch was applied by the blunt forceps at approximately 3 cm from the tip of tail. The reaction to tail pinch was observed and was documented.
All above four responses were graded as follows:
1 = No reaction.
2 = orientating response: Slowly turns towards the stimulus or walks away.
3 = Startle response or freezing reaction.
4 = More energetic response than “2” or “3”.
5 = Jumps at or away, attack or bites.
3) Visual Placing Response:
The animal was removed from its cage and held at the base of the tail (holding the tail more distally can strip off the skin) then slowly lowered forward towards the edge of the observation area or another raised edge (such as the rim of an overturned cage). The visual placing response is graded and documented:
1 = Early extension of forelimbs to reach for the screen.
2 = Extends limbs only after contact with the vibrissae or nose.
3 = No extension even after contact with nose.
4) Air righting response:
Holding the animal in a supine position, it was dropped from approximately 30cm and the righting response was rated:
1 = Lands with all feet on the ground.
2 = Uncoordinated landing or lands on side.
3 = Lands on back.
Grip strength: Grip strength of fore limbs was measured with a digital grip strength meter (Columbus Instruments International Corporation, Ohio, USA) to determine the ability of the rat to grasp and hold on the mesh platform. The grip strength of each rat was measured in Kilogram (Kg) for 3 consecutive times and average of the three grip strength values was calculated.
Motor activity: Motor activity of each animal was monitored using an automated animal activity measuring system (Columbus Instruments, OPTO-M3, Ohio, USA). Animals were monitored for three consecutive 10 minutes intervals allowing for examination of both exploratory and acclimation activity levels. During this period, total and ambulatory activity of the animal was recorded. Stereotypic activity was calculated by subtracting ambulatory activity from total activity.
Other: No Data - Postmortem examinations (parental animals):
- SACRIFICE: The rats were sacrificed by CO2 asphyxiation, after 28 consecutive days of oral administration, all surviving study rats were sacrificed on day 29 (Group I, III, IV, V).
GROSS PATHOLOGY: Yes, All the rats survived through the dosing period of 28 days and were sacrificed and gross lesions were noted.
ORGAN WEIGHTS: Liver, Kidneys, Adrenals, Epididymides, Prostate + Seminal Vesicle with Coagulation gland as whole, Thymus, Spleen, Brain, Heart, Lungs, Uterus, Testes/Ovaries were dissected free of fat and weighed. The paired organs were weighed together.
HISTOPATHOLOGY: Yes, From each rat, samples or the whole of the tissues listed below were preserved. All tissues were fixed in 10% neutral buffered formalin except, eyes and testes of all animals were preserved in Davidson’s solution for 24 hours and transferred to 10% neutral buffered formalin. Procedure for preparation of slides of tissues of various organs from the rats of various dose groups were performed.
Following tissue samples of organs from control and animals treated at different dose groups were preserved and those from control and treated at the highest dose level of 1000 mg/kg were subjected to histopathological examination:
Adrenals, Aorta, Brain (cerebrum, cerebellum and pons), Caecum, Cervix, Colon, Duodenum, Epididymides, Eyes, Heart, Ileum, Jejunum, Kidneys, Liver, Lungs, Mesenteric Lymphnodes, Oesophagus, Ovaries, Pancreas, Pituitary gland, Pharyngeal Lymphnodes, Prostate, Rectum, Skeletal Muscles, Skin with Mammary Gland, Spleen, Sternum with bone marrow, Sciatic Nerve, Spinal Cord (Cervical, mid thoracic and lumbar), Stomach, Seminal Vesicles with Coagulation Gland, Testes, Thymus, Thyroid, Trachea, Vagina, Urinary Bladder, Uterus. - Statistics:
- Raw data was processed and analyzed for reporting group means and standard deviations with significance between the controls and treated groups, using in-house developed and validated MS-Excel 2003 based statistical software. All the parameters characterized by continuous data such as body weight, per cent body weight change, feed consumption, organ weight, relative organ weight, haematological and clinical chemistry data were subjected to Bartlett’s test to meet the homogeneity of variance before conducting Analyses of Variance (ANOVA) and Dunnett’s t-test. Where the data did not meet the homogeneity of variance, Student’s t-test was performed to calculate significance.
Significance was calculated at 1% as well as 5% level and indicated in the summary tables as follows:
* = Significant than control at 95% level of confidence (p≤ 0.05).
** = Significant than control at 99% level of confidence (p≤ 0.01).
Results and discussion
Results: P0 (first parental generation)
General toxicity (P0)
- Clinical signs:
- no effects observed
- Description (incidence and severity):
- No clinical signs of toxicity were observed in the animals throughout the dosing period of 28 days
- Dermal irritation (if dermal study):
- not examined
- Mortality:
- no mortality observed
- Description (incidence):
- All the animals from control and all the treated dose groups survived throughout the dosing period of 28 days.
- Body weight and weight changes:
- no effects observed
- Description (incidence and severity):
- Animals from control and different dose groups exhibited normal body weight gain throughout the dosing period of 28 days.
- Food consumption and compound intake (if feeding study):
- no effects observed
- Description (incidence and severity):
- Animals from control and all treated dose groups exhibited normal feed consumption at the end of the dosing period of 28 days.
- Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- not examined
- Ophthalmological findings:
- not examined
- Haematological findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Haematological investigations conducted at the end of dosing period on day 29, revealed following significant changes in the values of different parameters studied when compared with that of respective controls.
Male :
MCV (p≤0.01), MCH (p≤0.01) and Total WBC (P≤0.05) : Increased values were obtained for animals from 250 mg/kg dose group,
Platelets : Increased values were obtained for animals from 250 mg/kg and 1000 mg/kg dose groups (p≤0.05),
Hb and HCT : Decreased values were obtained for animals from 250 mg/kg (p≤0.05), 500 mg/kg (p≤0.01) and 1000 mg/kg (p≤0.01) dose groups,
Total RBC : Decreased values were obtained for animals from 250 mg/kg and 500 mg/kg dose groups (p≤0.01) and
MCV and MCH : Decreased values were obtained for animals from 1000 mg/kg dose group (p≤0.01).
Female :
Platelets : Increased values were obtained for animals from 500 mg/kg dose group (p≤0.01),
Hb : Decreased values were obtained for animals from 250 mg/kg (p≤0.05) and 500 mg/kg (p≤0.01) dose groups,
Total RBC (p≤0.01), HCT (p≤0.05) and Total WBC (P≤0.05) : Decreased values were obtained for animals from 500 mg/kg dose group,
MCV : Decreased values were obtained for animals from 1000 mg/kg dose group (p≤0.01) and
MCH and MCHC : Decreased values were obtained for animals from 250 mg/kg (p≤0.05) and 1000 mg/kg (p≤0.01) dose groups.
The observed changes in hematology either lacked dose-dependency, consistency between sexes, or gross or histopathological correlations and were therefore considered incidental or non-adverse. - Clinical biochemistry findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Biochemical investigations conducted at the end of dosing period on day 29, revealed following significant changes in the values of different parameters studied when compared with that of respective controls.
Male :
Total Protein : Elevated levels were observed in animals from 500 mg/kg dose group (p≤0.05),
Creatinine : Elevated levels were observed in animals from 1000 mg/kg dose group (p≤0.05),
Cholesterol : Elevated levels were observed in animals from 500 mg/kg and 1000 mg/kg dose groups (p≤0.05),
Bilirubin : Decreased levels were observed in animals from 250 mg/kg (p≤0.05) and 500 mg/kg (p≤0.01) dose groups and
Chloride : Decreased levels were observed in animals from 500 mg/kg dose group (p≤0.01).
Female :
Sodium : Elevated levels were observed in animals from 250 mg/kg, 500 mg/kg and 1000 mg/kg dose groups (p≤0.01),
Alkaline Phosphatase : Decreased levels were observed in animals from 500 mg/kg dose group (p≤0.01) and
Chloride : Decreased levels were observed in animals from 1000 mg/kg dose group (p≤0.05).
The observed changes in clinical chemistry either lacked dose-dependency, consistency between sexes, or gross or histopathological correlations and were therefore considered incidental or non-adverse. - Urinalysis findings:
- not examined
- Behaviour (functional findings):
- effects observed, non-treatment-related
- Description (incidence and severity):
- Sensory Reactivity Observations:
All animals from control and different dose groups showed normal arousal level, visual response, touch response, auditory response, tail pinch response and visual placing response. Normal air righting reflex was observed in all animals from control and different dose groups in week 4.
Grip Strength:
Grip strength values observed in male and female animals for control and different dose groups were comparable.
Motor Activity:
Higher values were observed in male animals from 250 mg/kg, 500 mg/kg and 1000 mg/kg dose groups for second interval (p≤0.05). Lower values were observed in female animals from 1000 mg/kg dose group for first interval (p≤0.01) and for second interval (p≤0.05).
These changes were within laboratory range and were considered to be of no toxicological importance - Immunological findings:
- not examined
- Organ weight findings including organ / body weight ratios:
- effects observed, non-treatment-related
- Histopathological findings: non-neoplastic:
- no effects observed
- Description (incidence and severity):
- No treatment related histopathological changes were evident in male and female rats from control and high dose groups.
Histopathological examination revealed minimal focal to multifocal periportal mononuclear cell infiltration in the liver; minimal interstitial haemorrhages and/or tubular dilatation in the kidneys; minimal alveolar haemorrhages and/or alveolar histiocytosis in the lungs; minimal multifocal haemosiderosis and/or diffused congestion in spleen; minimal eosinophilic cells infiltration and/or luminal dilatation and/or endometrial gland dilatation in uterus; minimal luminal seminal coagulum in the urinary bladder; minimal dilatation of zona reticularis and/or vacuolation in zona fasiculata in adrenals; minimal multifocal haemorrhages in thymus; presence of ultimobranchial cysts in thyroid; in male or female animals from control and high dose group. All the changes observed in the control and high dose treatment group animals were similar, incidental and mode of death related, physiological and are covered in the facility historical data of the histopathology findings. - Histopathological findings: neoplastic:
- no effects observed
Effect levels (P0)
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 1 000 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- clinical signs
- mortality
- body weight and weight gain
- food consumption and compound intake
- haematology
- clinical biochemistry
- organ weights and organ / body weight ratios
- gross pathology
- histopathology: non-neoplastic
- histopathology: neoplastic
Target system / organ toxicity (P0)
- Critical effects observed:
- not specified
- Organ:
- not specified
Results: F1 generation
Effect levels (F1)
- Remarks on result:
- not measured/tested
Target system / organ toxicity (F1)
- Critical effects observed:
- no
Overall reproductive toxicity
- Reproductive effects observed:
- not specified
- Treatment related:
- not specified
Any other information on results incl. tables
VIABILITY
|
Dose |
Mortality |
||||
Group |
mg/kg |
Males |
Females |
|||
Number |
Male |
Female |
Absolute |
Relative % |
Absolute |
Relative % |
I |
0 |
0 |
0/6 |
0 |
0/6 |
0 |
II |
250 |
250 |
0/6 |
0 |
0/6 |
0 |
III |
500 |
500 |
0/6 |
0 |
0/6 |
0 |
IV |
1000 |
1000 |
0/6 |
0 |
0/6 |
0 |
GROUP MEAN BODY WEIGHT (g)
Sex - Male
Group |
Dose |
|
Day |
|||||
Number |
mg/kg bw |
|
0 |
1 |
8 |
15 |
22 |
28 |
I |
0 |
Mean |
154.23 |
159.17 |
214.55 |
266.05 |
299.30 |
323.63 |
±SD |
11.21 |
11.28 |
14.09 |
19.19 |
20.96 |
19.45 |
||
II |
250 |
Mean |
152.37 |
157.73 |
207.88 |
261.47 |
292.90 |
316.53 |
±SD |
12.54 |
12.53 |
12.50 |
15.07 |
14.41 |
14.22 |
||
III |
500 |
Mean |
153.25 |
159.88 |
201.80 |
248.33 |
285.63 |
306.90 |
±SD |
11.25 |
11.82 |
11.18 |
11.82 |
13.66 |
13.97 |
||
IV |
1000 |
Mean |
155.10 |
161.08 |
205.98 |
260.42 |
291.13 |
311.83 |
±SD |
9.84 |
9.88 |
7.48 |
16.30 |
12.05 |
10.81 |
Sex - Female
Group |
Dose |
|
Day |
|||||
Number |
mg/kg bw |
|
0 |
1 |
8 |
15 |
22 |
28 |
I |
0 |
Mean |
138.18 |
141.05 |
179.03 |
204.52 |
220.50 |
234.18 |
±SD |
9.51 |
9.69 |
8.67 |
10.74 |
11.85 |
11.00 |
||
II |
250 |
Mean |
138.95 |
142.18 |
169.42 |
192.18 |
210.32 |
224.70 |
±SD |
8.80 |
8.94 |
4.88 |
7.42 |
15.75 |
15.72 |
||
III |
500 |
Mean |
139.28 |
142.22 |
169.40 |
189.75 |
206.50 |
221.45 |
±SD |
9.47 |
9.97 |
5.81 |
5.78 |
12.09 |
12.83 |
||
IV |
1000 |
Mean |
140.22 |
143.20 |
169.85 |
187.55 |
202.20 |
218.48 |
±SD |
10.30 |
10.14 |
7.09 |
6.25 |
10.41 |
8.98 |
GROUP MEAN FEED CONSUMPTION (g/animal/day)
Sex - Male
Group |
Dose |
|
WEEK |
||||
Number |
mg/kg |
|
0 Day |
1 |
2 |
3 |
4 |
I |
0 |
Mean |
15.73 |
17.73 |
19.65 |
21.50 |
24.18 |
II |
250 |
Mean |
15.46 |
17.63 |
19.43 |
21.82 |
23.62 |
III |
500 |
Mean |
15.60 |
17.96 |
18.58 |
20.97 |
23.38 |
IV |
1000 |
Mean |
15.55 |
17.68 |
19.32 |
21.02 |
23.32 |
Sex - Female
Group |
Dose |
|
WEEK |
||||
Number |
mg/kg |
|
0 Day |
1 |
2 |
3 |
4 |
I |
0 |
Mean |
11.76 |
13.38 |
15.53 |
16.85 |
19.62 |
II |
250 |
Mean |
11.88 |
13.55 |
15.38 |
16.70 |
19.21 |
III |
500 |
Mean |
11.62 |
13.22 |
15.03 |
16.48 |
18.62 |
IV |
1000 |
Mean |
12.03 |
13.58 |
15.22 |
16.22 |
18.58 |
SUMMARY OF CLINICAL OBSERVATIONS
AND GENERAL APPEARANCE
Sex : Male
Group Number |
Dose mg/kg |
Observed Signs |
Total Number of Animals |
Animal Nos. |
Period of signs in days from - to |
Mortality |
I |
0 |
Nil |
6 |
1 - 6 |
1 - 28 |
0/6 |
II |
250 |
Nil |
6 |
13 - 18 |
1 - 28 |
0/6 |
III |
500 |
Nil |
6 |
25 - 30 |
1 - 28 |
0/6 |
IV |
1000 |
Nil |
6 |
37 - 42 |
1 - 28 |
0/6 |
Sex : Female
Group Number |
Dose mg/kg |
Observed Signs |
Total Number of Animals |
Animal Nos. |
Period of signs in days from - to |
Mortality |
I |
0 |
Nil |
6 |
7 - 12 |
1 - 28 |
0/6 |
II |
250 |
Nil |
6 |
19 - 24 |
1 - 28 |
0/6 |
III |
500 |
Nil |
6 |
31 - 36 |
1 - 28 |
0/6 |
IV |
1000 |
Nil |
6 |
43 - 48 |
1 - 28 |
0/6 |
GROUP MEAN HAEMATOLOGY
Sex : Male
Day : 29
Group |
Dose |
|
Hb |
Total RBC |
HCT |
MCV |
MCH |
MCHC |
Number |
mg/kg |
|
(g/dL) |
(x 106/µL) |
(%) |
(fL) |
(pg) |
(g/dL) |
I |
0 |
Mean |
17.62 |
8.05 |
43.97 |
54.63 |
21.92 |
40.07 |
±SD |
0.31 |
0.08 |
0.55 |
0.45 |
0.38 |
0.42 |
||
II |
250 |
Mean |
16.75* |
7.40** |
41.75* |
56.48** |
22.68** |
40.12 |
±SD |
0.48 |
0.28 |
1.31 |
0.79 |
0.18 |
0.52 |
||
III |
500 |
Mean |
15.62** |
7.21** |
39.40** |
54.77 |
21.72 |
39.68 |
±SD |
0.74 |
0.51 |
2.13 |
2.84 |
1.10 |
0.28 |
||
IV |
1000 |
Mean |
16.17** |
7.91 |
40.68** |
51.48** |
20.43** |
39.70 |
±SD |
0.47 |
0.33 |
1.27 |
0.89 |
0.46 |
0.40 |
Group |
Dose |
|
Platelets |
Total WBC |
Differential % |
Pt. |
||||
Number |
mg/kg |
|
(x 103/ µL) |
(x 103/µL) |
N |
L |
E |
M |
B |
(Sec.) |
I |
0 |
Mean |
305.33 |
8.08 |
19.33 |
78.00 |
1.67 |
1.00 |
0.00 |
14.67 |
±SD |
7.15 |
1.19 |
2.73 |
3.63 |
0.82 |
0.89 |
0.00 |
3.56 |
||
II |
250 |
Mean |
321.00* |
13.20* |
18.67 |
79.00 |
1.17 |
1.17 |
0.00 |
15.83 |
±SD |
15.58 |
4.02 |
2.34 |
1.90 |
1.17 |
0.75 |
0.00 |
2.86 |
||
III |
500 |
Mean |
341.50 |
7.92 |
18.67 |
79.00 |
1.33 |
1.00 |
0.00 |
15.67 |
±SD |
45.31 |
3.91 |
2.34 |
2.97 |
1.21 |
0.89 |
0.00 |
3.14 |
||
IV |
1000 |
Mean |
338.17* |
9.07 |
18.67 |
79.17 |
1.17 |
1.00 |
0.00 |
13.50 |
±SD |
32.50 |
1.77 |
2.16 |
2.79 |
0.75 |
0.89 |
0.00 |
1.87 |
Hb : Hemoglobin RBC : Red Blood Corpuscules
HCT : Hematocrit MCV : Mean Corpuscular Volume
MCH : Mean Corpuscular Hemoglobin MCHC : Mean Corpuscular Hemoglobin Concentration
WBC : White Blood Corpuscles Pt. : Prothrombin time
N : Neutrophils L : Lymphocytes
E : Eosinophils M : Monocytes
B : Basophils
* = Significant at 95% level of confidence (p≤0.05)
** = Significant at 99% level of confidence (p≤0.01)
Sex : Female
Day : 29
Group |
Dose |
|
Hb |
Total RBC |
HCT |
MCV |
MCH |
MCHC |
Number |
mg/kg |
|
(g/dL) |
(x 106/µL) |
(%) |
(fL) |
(pg) |
(g/dL) |
I |
0 |
Mean |
17.43 |
8.00 |
43.10 |
53.85 |
21.80 |
40.47 |
±SD |
0.31 |
0.30 |
1.14 |
0.72 |
0.49 |
0.41 |
||
II |
250 |
Mean |
16.40* |
8.07 |
41.30 |
51.32 |
20.40* |
39.78* |
±SD |
0.79 |
0.39 |
2.03 |
3.53 |
1.45 |
0.22 |
||
III |
500 |
Mean |
15.18** |
6.94** |
37.90* |
54.58 |
21.90 |
40.10 |
±SD |
0.54 |
0.25 |
1.43 |
0.37 |
0.17 |
0.54 |
||
IV |
1000 |
Mean |
16.63 |
8.12 |
42.25 |
52.07** |
20.48** |
39.38** |
±SD |
0.85 |
0.52 |
2.32 |
0.96 |
0.37 |
0.35 |
Group |
Dose |
|
Platelets |
Total WBC |
Differential % |
Pt. |
||||
Number |
mg/kg |
|
(x 103/ µL) |
(x 103/µL) |
N |
L |
E |
M |
B |
(Sec.) |
I |
0 |
Mean |
295.83 |
8.57 |
17.83 |
80.33 |
1.17 |
0.67 |
0.00 |
15.33 |
±SD |
30.90 |
2.43 |
2.32 |
2.34 |
0.75 |
0.52 |
0.00 |
3.08 |
||
II |
250 |
Mean |
308.17 |
11.80 |
17.83 |
79.50 |
1.67 |
1.00 |
0.00 |
16.33 |
±SD |
59.19 |
3.96 |
2.99 |
3.33 |
0.82 |
0.63 |
0.00 |
2.80 |
||
III |
500 |
Mean |
372.33** |
5.48* |
17.33 |
80.83 |
1.17 |
0.67 |
0.00 |
16.17 |
±SD |
18.48 |
0.17 |
2.88 |
2.48 |
0.75 |
0.52 |
0.00 |
2.79 |
||
IV |
1000 |
Mean |
318.00 |
8.33 |
17.67 |
79.83 |
1.00 |
1.50 |
0.00 |
15.33 |
±SD |
39.65 |
2.37 |
3.01 |
2.93 |
0.63 |
0.55 |
0.00 |
2.16 |
Hb : Hemoglobin RBC : Red Blood Corpuscules
HCT : Hematocrit MCV : Mean Corpuscular Volume
MCH : Mean Corpuscular Hemoglobin MCHC : Mean Corpuscular Hemoglobin Concentration
WBC : White Blood Corpuscles Pt. : Prothrombin time
N : Neutrophils L : Lymphocytes
E : Eosinophils M : Monocytes
B : Basophils
* = Significant at 95% level of confidence (p≤0.05)
** = Significant at 99% level of confidence (p≤0.01)
GROUP MEAN CLINICAL BIOCHEMISTRY
Sex : Male
Day : 29
Group Number |
Dose mg/kg |
|
Total Protein (g/dL) |
BUN (mg/dL) |
Urea Nitrogen (mg/dL) |
ALT (U/L) |
AST (U/L) |
ALP (U/L) |
Glucose (mg/dL) |
I |
0 |
Mean |
6.57 |
14.17 |
30.88 |
48.17 |
101.50 |
171.50 |
74.33 |
±SD |
0.29 |
1.72 |
3.75 |
3.25 |
17.95 |
37.13 |
6.53 |
||
II |
250 |
Mean |
6.77 |
16.00 |
34.88 |
46.67 |
92.17 |
172.67 |
76.50 |
±SD |
0.29 |
1.26 |
2.76 |
7.45 |
13.12 |
34.20 |
8.92 |
||
III |
500 |
Mean |
7.04* |
15.17 |
33.06 |
48.17 |
104.33 |
185.50 |
78.50 |
±SD |
0.19 |
1.47 |
3.21 |
9.87 |
24.20 |
33.12 |
5.21 |
||
IV |
1000 |
Mean |
6.77 |
14.83 |
32.34 |
48.83 |
104.00 |
193.67 |
65.33 |
±SD |
0.28 |
2.48 |
5.41 |
4.79 |
8.46 |
22.26 |
5.43 |
Group Number |
Dose mg/kg |
|
Calcium (mmol/L) |
Phospho-rous (mg/dL) |
GGT (U/L) |
Total Bilirubin (mg/dL) |
Albumin (g/dL) |
Globulin (g/dL) |
Creatinine (mg/dL) |
I |
0 |
Mean |
3.59 |
7.62 |
6.50 |
0.31 |
1.17 |
5.40 |
0.45 |
±SD |
0.11 |
0.83 |
1.22 |
0.04 |
0.13 |
0.39 |
0.03 |
||
II |
250 |
Mean |
3.69 |
7.48 |
6.33 |
0.20* |
1.28 |
5.48 |
0.46 |
±SD |
0.12 |
0.88 |
0.52 |
0.09 |
0.09 |
0.29 |
0.05 |
||
III |
500 |
Mean |
3.76 |
8.03 |
5.17 |
0.13** |
1.21 |
5.82 |
0.47 |
±SD |
0.10 |
0.89 |
0.75 |
0.02 |
0.09 |
0.17 |
0.04 |
||
IV |
1000 |
Mean |
3.68 |
8.27 |
5.67 |
0.27 |
1.27 |
5.48 |
0.52* |
±SD |
0.10 |
0.82 |
0.82 |
0.02 |
0.10 |
0.25 |
0.04 |
Group Number |
Dose mg/kg |
|
Sodium (mmol/L) |
Potassium (mmol/L) |
Chloride (mmol/L) |
Total Cholesterol (mg/dL) |
Triglycerides (mg/dL) |
I |
0 |
Mean |
146.34 |
4.22 |
109.07 |
31.33 |
74.17 |
±SD |
1.30 |
0.19 |
1.26 |
8.31 |
20.54 |
||
II |
250 |
Mean |
145.43 |
4.54 |
107.77 |
41.00 |
66.67 |
±SD |
0.80 |
0.26 |
1.67 |
7.16 |
14.46 |
||
III |
500 |
Mean |
146.76 |
4.67 |
105.66** |
43.33* |
81.17 |
±SD |
1.46 |
0.43 |
1.90 |
6.38 |
23.66 |
||
IV |
1000 |
Mean |
146.40 |
4.23 |
108.67 |
42.83* |
77.33 |
±SD |
1.91 |
0.32 |
1.01 |
8.01 |
30.87 |
BUN : Blood Urea Nitrogen ALT : Alanine Aminotransferase AST : Aspartate Transaminase ALP : Alkaline Phosphatase |
|
* = Significant at 95% level of confidence (p≤0.05)
** = Significant at 99% level of confidence (p≤0.01)
GROUP MEAN CLINICAL BIOCHEMISTRY
Laboratory Test Item Code : TAS/122/001
Test System : Sprague Dawley Rat
Sex : Female
Day : 29
Group Number |
Dose mg/kg |
|
Total Protein (g/dL) |
BUN (mg/dL) |
Urea Nitrogen (mg/dL) |
ALT (U/L) |
AST (U/L) |
ALP (U/L) |
Glucose (mg/dL) |
I |
0 |
Mean |
6.53 |
15.17 |
33.06 |
44.83 |
107.33 |
164.00 |
72.83 |
±SD |
0.31 |
2.23 |
4.86 |
4.31 |
15.58 |
20.07 |
4.58 |
||
II |
250 |
Mean |
6.48 |
15.17 |
33.06 |
45.50 |
113.33 |
132.00 |
64.00 |
±SD |
0.39 |
1.60 |
3.49 |
9.03 |
23.59 |
30.57 |
8.92 |
||
III |
500 |
Mean |
6.53 |
13.67 |
29.79 |
42.50 |
90.83 |
99.00** |
70.50 |
±SD |
0.43 |
0.82 |
1.78 |
6.60 |
8.98 |
20.42 |
6.41 |
||
IV |
1000 |
Mean |
6.42 |
15.33 |
33.43 |
41.50 |
95.67 |
125.83 |
71.67 |
±SD |
0.44 |
3.72 |
8.12 |
5.86 |
9.07 |
33.47 |
7.45 |
Group Number |
Dose mg/kg |
|
Calcium (mmol/L) |
Phospho-rous (mg/dL) |
GGT (U/L) |
Total Bilirubin (mg/dL) |
Albumin (g/dL) |
Globulin (g/dL) |
Creatinine (mg/dL) |
I |
0 |
Mean |
3.71 |
6.57 |
6.50 |
0.11 |
1.29 |
5.25 |
0.55 |
±SD |
0.11 |
0.64 |
1.05 |
0.02 |
0.15 |
0.29 |
0.05 |
||
II |
250 |
Mean |
3.71 |
7.48 |
6.67 |
0.13 |
1.19 |
5.32 |
0.53 |
±SD |
0.11 |
0.75 |
1.37 |
0.06 |
0.17 |
0.33 |
0.04 |
||
III |
500 |
Mean |
3.68 |
7.17 |
5.17 |
0.12 |
1.29 |
5.23 |
0.51 |
±SD |
0.04 |
0.84 |
0.75 |
0.02 |
0.10 |
0.31 |
0.05 |
||
IV |
1000 |
Mean |
3.71 |
6.77 |
6.33 |
0.11 |
1.22 |
5.20 |
0.55 |
±SD |
0.04 |
0.94 |
1.75 |
0.02 |
0.13 |
0.40 |
0.06 |
Group Number |
Dose mg/kg |
|
Sodium (mmol/L) |
Potassium (mmol/L) |
Chloride (mmol/L) |
Total Cholesterol (mg/dL) |
Triglycerides (mg/dL) |
I |
0 |
Mean |
144.61 |
4.34 |
108.89 |
52.33 |
76.00 |
±SD |
0.66 |
0.40 |
1.06 |
5.09 |
15.49 |
||
II |
250 |
Mean |
146.38** |
4.43 |
107.33 |
48.17 |
63.17 |
±SD |
0.77 |
0.22 |
2.43 |
8.91 |
19.01 |
||
III |
500 |
Mean |
146.32** |
4.25 |
108.89 |
47.83 |
83.33 |
±SD |
0.54 |
0.14 |
1.53 |
7.88 |
25.73 |
||
IV |
1000 |
Mean |
146.45** |
4.21 |
106.32* |
50.17 |
88.00 |
±SD |
1.42 |
0.27 |
1.21 |
5.85 |
28.71 |
BUN : Blood Urea Nitrogen ALT : Alanine Aminotransferase AST : Aspartate Transaminase ALP : Alkaline Phosphatase |
|
* = Significant at 95% level of confidence (p≤0.05)
** = Significant at 99% level of confidence (p≤0.01)
GROUP MEAN ABSOLUTE ORGAN WEIGHTS (g)
Sex : Male
Day : 29
GroupNumber |
Dose mg/kg |
|
Body Weight (g) |
Brain |
Liver
|
Kidneys
|
Adrenals
|
Testes |
Prostate + Seminal Vesicle with Coagulation gland |
I |
0 |
Mean |
304.27 |
2.033 |
13.192 |
2.429 |
0.0607 |
3.182 |
1.491 |
±SD |
19.75 |
0.144 |
1.775 |
0.195 |
0.0133 |
0.139 |
0.284 |
||
II |
250 |
Mean |
298.33 |
1.936 |
12.352 |
2.244 |
0.0545 |
3.077 |
1.258 |
±SD |
15.62 |
0.154 |
1.338 |
0.202 |
0.0028 |
0.318 |
0.154 |
||
III |
500 |
Mean |
288.80 |
1.975 |
12.340 |
2.162 |
0.0523 |
2.594 |
1.133 |
±SD |
13.01 |
0.078 |
0.903 |
0.409 |
0.0067 |
0.276 |
0.246 |
||
IV |
1000 |
Mean |
294.33 |
1.999 |
12.163 |
2.347 |
0.0474 |
2.964 |
1.213 |
±SD |
10.28 |
0.086 |
1.233 |
0.214 |
0.0070 |
0.249 |
0.122 |
GroupNumber |
Dose mg/kg |
|
Heart |
Spleen |
Lungs |
Thymus |
Epididymides |
I |
0 |
Mean |
1.364 |
1.404 |
1.760 |
0.430 |
0.764 |
±SD |
0.162 |
0.247 |
0.238 |
0.100 |
0.058 |
||
II |
250 |
Mean |
1.231 |
1.349 |
1.623 |
0.372 |
0.671 |
±SD |
0.045 |
0.413 |
0.188 |
0.068 |
0.048 |
||
III |
500 |
Mean |
1.233 |
1.411 |
1.817 |
0.363 |
0.658 |
±SD |
0.072 |
0.227 |
0.441 |
0.066 |
0.032 |
||
IV |
1000 |
Mean |
1.133 |
1.488 |
1.531 |
0.343 |
0.673 |
±SD |
0.144 |
0.513 |
0.315 |
0.081 |
0.076 |
GROUP MEAN ABSOLUTE ORGAN WEIGHTS (g)
Sex : Female
Day : 29
GroupNumber |
Dose mg/kg |
|
Body Weight (g) |
Brain |
Liver
|
Kidneys
|
Adrenals
|
Ovaries
|
I |
0 |
Mean |
217.20 |
1.908 |
8.040 |
1.646 |
0.0559 |
0.0822 |
±SD |
11.85 |
0.098 |
0.889 |
0.136 |
0.0087 |
0.0105 |
||
II |
250 |
Mean |
207.97 |
1.849 |
7.651 |
1.475 |
0.0556 |
0.0669 |
±SD |
16.31 |
0.111 |
1.027 |
0.176 |
0.0078 |
0.0095 |
||
III |
500 |
Mean |
206.17 |
1.813 |
7.788 |
1.523 |
0.0558 |
0.0757 |
±SD |
11.88 |
0.124 |
1.110 |
0.261 |
0.0044 |
0.0192 |
||
IV |
1000 |
Mean |
201.28 |
1.828 |
7.225 |
1.478 |
0.0512 |
0.0703 |
±SD |
8.75 |
0.069 |
0.878 |
0.150 |
0.0082 |
0.0076 |
Group No. |
Dose mg/kg |
|
Heart |
Spleen |
Lungs |
Thymus |
Uterus |
I |
0 |
Mean |
0.955 |
1.130 |
1.394 |
0.435 |
0.346 |
±SD |
0.085 |
0.271 |
0.361 |
0.097 |
0.077 |
||
II |
250 |
Mean |
0.881 |
1.396 |
1.462 |
0.386 |
0.313 |
±SD |
0.088 |
0.605 |
0.204 |
0.147 |
0.106 |
||
III |
500 |
Mean |
0.848 |
1.028 |
1.632 |
0.363 |
0.293 |
±SD |
0.113 |
0.198 |
0.181 |
0.085 |
0.079 |
||
IV |
1000 |
Mean |
0.815 |
0.834 |
1.322 |
0.345 |
0.293 |
±SD |
0.048 |
0.149 |
0.217 |
0.050 |
0.150 |
GROUP MEAN RELATIVE ORGAN WEIGHTS (%)
Sex : Male
Day : 29
GroupNumber |
Dose mg/kg |
|
Body Weight (g) |
Brain |
Liver
|
Kidneys
|
Adrenals
|
Testes |
Prostate + Seminal Vesicle with Coagulation gland |
I |
0 |
Mean |
304.27 |
0.668 |
4.326 |
0.798 |
0.0199 |
1.048 |
0.490 |
±SD |
19.75 |
0.029 |
0.404 |
0.034 |
0.0039 |
0.043 |
0.082 |
||
II |
250 |
Mean |
298.33 |
0.650 |
4.138 |
0.753 |
0.0183 |
1.033 |
0.423 |
±SD |
15.62 |
0.055 |
0.368 |
0.066 |
0.0012 |
0.120 |
0.057 |
||
III |
500 |
Mean |
288.80 |
0.685 |
4.280 |
0.752 |
0.0182 |
0.899* |
0.393 |
±SD |
13.01 |
0.036 |
0.381 |
0.153 |
0.0025 |
0.099 |
0.085 |
||
IV |
1000 |
Mean |
294.33 |
0.680 |
4.131 |
0.798 |
0.0161 |
1.008 |
0.412 |
±SD |
10.28 |
0.031 |
0.379 |
0.072 |
0.0026 |
0.083 |
0.041 |
GroupNumber |
Dose mg/kg |
|
Heart |
Spleen |
Lungs |
Thymus |
Epididymides |
I |
0 |
Mean |
0.448 |
0.461 |
0.578 |
0.141 |
0.251 |
±SD |
0.042 |
0.068 |
0.063 |
0.031 |
0.015 |
||
II |
250 |
Mean |
0.413 |
0.448 |
0.546 |
0.125 |
0.225 |
±SD |
0.014 |
0.115 |
0.080 |
0.022 |
0.018 |
||
III |
500 |
Mean |
0.428 |
0.491 |
0.631 |
0.126 |
0.228 |
±SD |
0.041 |
0.095 |
0.162 |
0.025 |
0.014 |
||
IV |
1000 |
Mean |
0.385 |
0.505 |
0.521 |
0.116 |
0.228 |
±SD |
0.048 |
0.173 |
0.110 |
0.026 |
0.022 |
*= Significant at 95% level of confidence
GROUP MEAN RELATIVE ORGAN WEIGHTS (%)
Sex : Female
Day : 29
GroupNumber |
Dose mg/kg |
|
Body Weight (g) |
Brain |
Liver
|
Kidneys
|
Adrenals
|
Ovaries
|
I |
0 |
Mean |
217.20 |
0.880 |
3.695 |
0.758 |
0.0257 |
0.0378 |
±SD |
11.85 |
0.047 |
0.262 |
0.038 |
0.0031 |
0.0047 |
||
II |
250 |
Mean |
207.97 |
0.892 |
3.672 |
0.709 |
0.0270 |
0.0324 |
±SD |
16.31 |
0.060 |
0.323 |
0.055 |
0.0049 |
0.0055 |
||
III |
500 |
Mean |
206.17 |
0.879 |
3.766 |
0.736 |
0.0272 |
0.0368 |
±SD |
11.88 |
0.032 |
0.372 |
0.095 |
0.0029 |
0.0094 |
||
IV |
1000 |
Mean |
201.28 |
0.910 |
3.588 |
0.734 |
0.0253 |
0.0349 |
±SD |
8.75 |
0.053 |
0.396 |
0.066 |
0.0032 |
0.0030 |
Group No. |
Dose mg/kg |
|
Heart |
Spleen |
Lungs |
Thymus |
Uterus |
I |
0 |
Mean |
0.439 |
0.519 |
0.641 |
0.201 |
0.160 |
±SD |
0.022 |
0.118 |
0.159 |
0.047 |
0.041 |
||
II |
250 |
Mean |
0.423 |
0.673 |
0.709 |
0.183 |
0.149 |
±SD |
0.024 |
0.304 |
0.131 |
0.057 |
0.041 |
||
III |
500 |
Mean |
0.410 |
0.496 |
0.792 |
0.177 |
0.142 |
±SD |
0.041 |
0.081 |
0.089 |
0.045 |
0.038 |
||
IV |
1000 |
Mean |
0.405 |
0.413 |
0.654 |
0.171 |
0.146 |
±SD |
0.015 |
0.060 |
0.084 |
0.021 |
0.076 |
SUM
MARY OF GROSS PATHOLOGY FINDINGS
Sex : Male
Site and lesion observed |
Group |
I |
II |
III |
IV |
Dose (mg/kg) |
0 |
250 |
500 |
1000 |
|
No Abnormality Detected
|
1 - 6 |
13 - 18 |
25 - 30 |
37 - 42 |
SUMMARY OF GROSS PATHOLOGY FINDINGS
Sex : Female
Site and lesion observed |
Group |
I |
II |
III |
IV |
Dose (mg/kg) |
0 |
250 |
500 |
1000 |
|
No Abnormality Detected
|
7 - 12 |
19 - 24 |
31 - 36 |
43 - 48 |
Applicant's summary and conclusion
- Conclusions:
- The No Observed Adverse Effect Level (NOAEL) for the test chemical in Sprague Dawley Rats, via oral gavage, for 28 days was considered to be 1000 mg/kg bw/day.
- Executive summary:
The Reproductive organ toxicity study for 28 -days by oral route was designed and conducted to determine the reproductive toxicity profile of the test chemical when administered daily for 28 days in the Sprague Dawley rats. The test chemical was dissolved in Polyethylene glycol and used at dose level of 0, 250 mg/kg, 500 mg/kg and 1000 mg/kg body weight. During the study period, the treated animals were observed or mortality, cllinical signs, body weight and food consumption changes, hematology and clinical chemistry, neurobehaviour, urinalysis, ophthalmology and were subjected to gross and histopathology. All the male and female animals from control and all the treated dose groups up to 1000 mg/kg survived throughout the dosing period of 28 days. Male and female animals from control and all the treated dose groups exhibited normal body weight gain and food intake at the end of the dosing period of 28 days. Daily and detailed (weekly) clinical observations did not reveal any signs of toxicity in male and female animals from different dose groups during the dosing period of 28 days. Towards the end of the exposure period in week 4, functional observation battery such as sensory reactivity to stimuli of different types (e.g. auditory, visual and proprioceptive stimuli) revealed no abnormalities attributable to the treatment. Grip strength values observed in male and female animals for control and different dose groups were comparable. Higher values for motor activity were observed in male animals from 250 mg/kg, 500 mg/kg and 1000 mg/kg dose groups for second interval. Lower values for motor activity were observed in female animals from 1000 mg/kg dose group for first and second interval. These changes were within laboratory range and were considered to be of no toxicological importance. Haematological analysis performed on 29thday revealed statistically significant increase in the values of MCV, MCH and Total WBC of male rats dosed at 250 mg/kg, increase values of Platelets of male rats dosed at 250 mg/kg and 1000 mg/kg and increased values of Platelets of female rats dosed at 500 mg/kg. In addition, statistically significant decrease was observed in the values of Hb and HCT of male rats dosed at 250 mg/kg, 500 mg/kg and 1000 mg/kg, decreased values of Total RBC of male rats dosed at 250 mg/kg and 500 mg/kg, decreased values of MCV and MCH of male rats dosed at 1000 mg/kg, decreased values of Hb of female rats dosed at 250 mg/kg and 500 mg/kg, decreased values of Total RBC, HCT and Total WBC of female rats dosed at 500 mg/kg, decreased values of MCV of female rats dosed at 1000 mg/kg, decreased values of MCH and MCHC of female rats dosed at 250 mg/kg and 1000 mg/kg, the increase / decrease in the values of different parameters were marginal and within the normal laboratory limits. Statistically significant increase of Total Protein (in male rats at 500 mg/Kg/day), Creatinine (in male rats at 1000 mg/Kg/day), Cholesterol (in male rats at 500 and 1000 mg/Kg/day), Sodium (in female rats at 250, 500 and 1000 mg/Kg/day) and statistically significant decrease in Bilirubin (in male rats at 250 and 500 mg/Kg/day), Chloride and Alkaline Phosphatase (in female rats at 500 mg/Kg/day) was observed. Although there was an increase/decrease in the values of various biochemical parameters, the deviations were marginal and within the range of normal laboratory limits. Organ weight data of male animals sacrificed on day 29, revealed decreased relative weights of testes of animals from 500 mg/kg dose group. Although significant changes in organ weights were observed in male animals from intermediate dose group, no related gross pathological or histological changes were seen and hence considered to be of no toxicological importance. Organ weight data of female animals sacrificed on day 29, was found to be comparable with that of controls. Gross pathological examination conducted in rats of all dose groups during necropsy did not reveal any abnormality attributable to the treatment. Histopathological examination conducted in rats of control and high dose groups did not reveal any abnormality attributable to the treatment. Hence based on the observations made, the No Observed Adverse Effect Level (NOAEL) for the test chemical in Sprague Dawley Rats, via oral gavage, for 28 days was considered to be 1000 mg/kg bw/day.
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