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EC number: 284-716-0 | CAS number: 84962-20-9
- 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
Developmental toxicity / teratogenicity
Administrative data
- Endpoint:
- developmental toxicity
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 18 July 2018 to 19 December 2018
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Remarks:
- OECD 414, adopted on 25 June 2018
- Justification for type of information:
- TESTING PROPOSAL ON VERTEBRATE ANIMALSTESTING PROPOSAL ON VERTEBRATE ANIMALS
NON-CONFIDENTIAL NAME OF SUBSTANCE:
- Name of the substance on which testing is proposed to be carried out: Hordaphos MDGB
CONSIDERATIONS THAT THE GENERAL ADAPTATION POSSIBILITIES OF ANNEX XI OF THE REACH REGULATION ARE NOT ADEQUATE TO GENERATE THE NECESSARY INFORMATION [please address all points below]:
- Available GLP studies: OECD 422
FURTHER INFORMATION ON TESTING PROPOSAL IN ADDITION TO INFORMATION PROVIDED IN THE MATERIALS AND METHODS SECTION:
- Details on study design: The test item, Hordaphos MDGB, was administered by the oral route at 10 mL/kg body weight to mated and presumed-pregnant females from Gestation Day [GD] 5 to 19.TESTING PROPOSAL ON VERTEBRATE ANIMALS
NON-CONFIDENTIAL NAME OF SUBSTANCE:
- Name of the substance on which testing is proposed to be carried out: Hordaphos MDGB
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 019
- Report date:
- 2019
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 414 (Prenatal Developmental Toxicity Study)
- Version / remarks:
- adopted on 25 June 2018
- Deviations:
- no
- Principles of method if other than guideline:
- Not applicable
- GLP compliance:
- yes (incl. QA statement)
- Limit test:
- no
Test material
- Reference substance name:
- Phosphoric acid, mixed esters with Bu alc. and ethylene glycol
- EC Number:
- 284-716-0
- EC Name:
- Phosphoric acid, mixed esters with Bu alc. and ethylene glycol
- Cas Number:
- 84962-20-9
- IUPAC Name:
- Reaction mass of Phosphoric acid mono-(2-hydroxy-ethyl) ester and Phosphoric acid monobutyl ester and Phosphoric acid bis-(2-hydroxy-ethyl) ester
- Reference substance name:
- Phosphoric acid, mixed with Ethylene glycol and Butanol
- IUPAC Name:
- Phosphoric acid, mixed with Ethylene glycol and Butanol
- Test material form:
- other: liquid
Constituent 1
Constituent 2
- Specific details on test material used for the study:
- SOURCE OF TEST MATERIAL
- Source and batch No.of test material: Clariant Plastics and Coatings (Deutschland) GmbH and DEH8006819
- Expiration date of the batch: 28 October 2019
- Purity test date: 01 March 2018
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Ambient (21 to 29°C)
- Stability under test conditions: Stable for 6 hours at Ambient (21 to 29°C)
- Solubility and stability of the test substance in the solvent/vehicle: The test item was clearly miscible with distilled water at the concentration of 35 mg/mL (highest dose concentration) and stable for 6 hours at Ambient (21 to 29°C).
TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Formulated
FORM AS APPLIED IN THE TEST: Liquid
Test animals
- Species:
- rat
- Strain:
- Sprague-Dawley
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: In-house bred
- Age at study initiation: 10 to 11 weeks
- Weight at study initiation: 200 to 300 g
- Fasting period before study: No
- Housing: Mating - three rats (one male and two females) were housed in standard polypropylene cages.
- Post mating - After confirming presence of sperm in the vaginal smear (Day 0 of pregnancy), the mated pairs were separated. Males were housed with their former cage mates while females were housed individually in polypropylene cages.
- Diet (ad libitum): Altromin Maintenance diet for rats and mice 1324 manufactured by Altromin Spezialfutter GmbH & Co. KG
- Water (ad libitum): Deep bore-well water passed through reverse osmosis unit
- Acclimation period: minimum period of five days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.5 to 22.9o C
- Humidity (%): 45 to 67%
- Air changes (per hr): 12 to 15
- Photoperiod (hrs dark / hrs light): 12 hours fluorescent light and 12 hours dark cycle
IN-LIFE DATES: From: 18 July 2018; To: 30 August 2018
Administration / exposure
- Route of administration:
- oral: gavage
- Vehicle:
- water
- Remarks on MMAD:
- Not applicable
- Details on exposure:
- PREPARATION OF DOSING SOLUTIONS:
VEHICLE
- Justification for use and choice of vehicle : The test item was clearly miscible with distilled water at the concentration of 35 mg/mL (highest dose concentration). Hence, distilled water was selected as a vehicle for test item formulation preparations.
- Concentration in vehicle: G1: 0 mg/mL; G2: 5 mg/mL; G3: 13 mg/mL; G4: 35 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg body weight
- Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- Dose formulation analysis for dose concentration verification was done by Analytical Chemistry department of Bioneeds India Private Limited. Sampling and analysis of formulations was performed during first week and last week (since treatment was from GD 5 to 19 only two treatment weeks has been considered) of the treatment. The samples were collected in duplicates (60 mL each) from vehicle control, low, mid and high dose concentrations
The collected samples were transferred to Analytical Chemistry department of Bioneeds India Private Limited for dose concentration analysis. One set of aliquot of each formulation was analyzed. The second aliquot was stored as a backup purpose at established stability conditions. The second set of samples were discarded, as the analysis results of first set of samples were within the limits. - Details on mating procedure:
- - Impregnation procedure: [cohoused]
- M/F ratio per cage: 1:2
- Length of cohabitation: 14 days
- After 14 days of unsuccessful pairing replacement of first male by another male with proven fertility.
- Further matings after two unsuccessful attempts: no
- Verification of same strain and source of both sexes: yes
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy - Duration of treatment / exposure:
- Gestation day 5 to 19 for each presumed pregnant female
- Frequency of treatment:
- Once daily
- Duration of test:
- 5 months
Doses / concentrationsopen allclose all
- Dose / conc.:
- 0 mg/kg bw/day
- Remarks:
- Vehicle control group
- Dose / conc.:
- 50 mg/kg bw/day
- Remarks:
- Low dose group
- Dose / conc.:
- 130 mg/kg bw/day
- Remarks:
- Mid dose group
- Dose / conc.:
- 350 mg/kg bw/day
- Remarks:
- High dose group
- No. of animals per sex per dose:
- 25 presumed pregnant females
- Control animals:
- yes, concurrent vehicle
- Details on study design:
- - Dose selection rationale:
The doses of 50, 130 and 350 mg/kg body weight for low, mid and high dose were selected as the dose range finding study (100, 300 and 1000 mg/kg body weight/day; duration: 14 days pre-mating, 14 days mating and gestation days 0 to 19) revealed mortality within the first week of dosing at 1000 mg/kg body weight/day. An abnormal breathing and attenuated body weight gain was reported at 300 mg/kg body weight/day. The number of implantation sites was slightly reduced in all groups. No treatment related observations were made in the course of visceral and skeletal examination.
The result of a subsequently performed OECD 422 study was observed with mortality beginning in week 2 of treatment, tracheal changes most probably related to local irritation at 200 mg/kg. The signs of discomfort (piloerection and abnormal breathing), tendency to decreased food consumption at 50 mg/kg was noted. The reproductive performance and developmental parameters were not affected in any dose group.
Based on the above mentioned observations the doses of 50, 130 and 350 mg/kg body weight for low, mid and high dose were selected
- Rationale for animal assignment: The body weight of mated females on each day of gestation day ‘0’ was recorded and arranged in the ascending order of their body weight until required number of mated females acquired for each group. These mated females were evenly distributed to all the groups based on their body weights so as to maintain comparable mean body weight for all groups and permanent identification numbers were assigned
Examinations
- Maternal examinations:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Once daily
- Cage side observations checked in table [No.1] were included.
DETAILED CLINICAL OBSERVATIONS: No
BODY WEIGHT: Yes
- Time schedule for examinations: gestation days (GD) 0, 3, 5, 8, 11, 14, 17, 19 and on 20 (day of caesarean section)
FOOD CONSUMPTION: Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
Individual animal feed intake of mated females was recorded for gestation days
0 to 3, 3 to 5, 5 to 8, 8 to 11, 11 to 14, 14 to 17, 17 to 19 and 19 to 20
POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 20
- Organs examined: All organs grossly with special attention to uterus, ovaries and thyroid - Ovaries and uterine content:
- The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes - Fetal examinations:
- - External examinations: Yes: [all per litter]
- Soft tissue examinations: Yes: [half per litter]
- Skeletal examinations: Yes: [half per litter]
- Head examinations: Yes: [half per litter] - Statistics:
- Data was subjected to various statistical analysis using SPSS software version 22. All analyses and comparisons were evaluated at the 95% level of confidence (P<0.05),
Data of non-pregnant females was not included in mean calculation and statistical analysis.
One-way ANOVA with Dunnett’s post test - Gestation body weight (g), Percent change in gestation
body weight, Corrected body weight (g & %), Gravid uterus weight (g), Feed consumption (g), Fetal
weights (g), Anogenital distance of all live fetuses (mm), Crown-rump length of fetuses (mm), serum
T4, T3 and TSH lelvels.
Kruskal-Wallis (Non-parametric) - No. of corpora lutea, implantations, resorptions, litter size, sex ratio, implantation lossess, live/dead fetuses.
Cross Tabs Chi-square test/ Fischer's Exact Test - Pregnancy rate, No. of dams with/without live/dead fetuses, No. of dams with/without resorptions - Indices:
- Corrected Body weight (g) = (Gestation day 20 body weight - Gestation day 5 body weight) - Gravid uterus weight.
Percent of Live/Dead Fetuses = (Number of Live/Dead Fetuses / Litter Size) x 100.
Percent of Early/Late Resorptions (%) = (Number of Early/late Resorptions/ Number of Implantations) x 100.
Early Resorptions per Group (%) = (Number of Dams with Early Resorptions in the group / Total Number of Dams in the Group) x 100.
Pre-implantation Loss (%) = (Number of Corpora lutea - Number of implants) / Number of Corporalutea x 100.
Post-implantation Loss (%) per Dam = (Number of Dams with Post-implantation loss / Total Number of Dams) x 100
Male/Female Sex Ratio = Number of live male fetuses / Number of live female fetuses.
Male/Female Fetuses (%) = (Number of live male/female fetuses / Total number of live fetuses) x 100.
Group/Litter Fetal Observation for External, Visceral and Skeletal Examinations:
Fetal incidence (%) = (Number of Fetuses with a particular observation / Total number of Fetuses in a group) x 100 - Historical control data:
- Not applicable
Results and discussion
Results: maternal animals
General toxicity (maternal animals)
- Clinical signs:
- no effects observed
- Description (incidence and severity):
- There were no clinical signs of toxicity and no mortality/morbidity observed at any of the tested dose group animals during the experimental period.
- Dermal irritation (if dermal study):
- not specified
- Description (incidence and severity):
- not applicable
- Mortality:
- no mortality observed
- Description (incidence):
- There were no mortality/morbidity observed at any of the tested dose group animals during the experimental period.
- Body weight and weight changes:
- no effects observed
- Description (incidence and severity):
- There were no changes noted in body weight during gestation period across all the tested dose groups when compared with the vehicle control group.
- Food consumption and compound intake (if feeding study):
- effects observed, non-treatment-related
- Description (incidence and severity):
- A statistically significant increase in average feed consumption during gestation day 3 to 5 in G4 group animals and statistically significant increase in average feed consumption during gestation day 17 to 19 and 19 to 20 in G2 group animals was observed when compared with vehicle control group. This sporadic incidence is considered as incidental and unrelated to treatment with test item due to unaffected body weights during this period
- Food efficiency:
- not examined
- Description (incidence and severity):
- not applicable
- Water consumption and compound intake (if drinking water study):
- not examined
- Description (incidence and severity):
- not applicable
- Ophthalmological findings:
- not examined
- Description (incidence and severity):
- not applicable
- Haematological findings:
- not examined
- Description (incidence and severity):
- not applicable
- Clinical biochemistry findings:
- not examined
- Description (incidence and severity):
- not applicable
- Urinalysis findings:
- not examined
- Description (incidence and severity):
- not applicable
- Behaviour (functional findings):
- not examined
- Description (incidence and severity):
- not applicable
- Immunological findings:
- not examined
- Description (incidence and severity):
- not applicable
- Organ weight findings including organ / body weight ratios:
- no effects observed
- Description (incidence and severity):
- The absolute thyroid along with parathyroid weights (g) were 0.0232, 0.0220, 0.0236 and 0.0235 and relative thyroid along with parathyroid weights (%) were 0.0062, 0.0058, 0.0063 and 0.0062 for G1 (0 mg/kg), G2 (50 mg/kg), G3 (130 mg/kg) and G4 (350 mg/kg) respectively. There were no statistically significant differences in mean absolute and relative thyroid along with parathyroid weight and no treatment related changes for this parameter across the dose groups when compared to controls
- Gross pathological findings:
- no effects observed
- Description (incidence and severity):
- There were no gross pathological changes noted in any of the dose group and vehicle control group animals during conduct of the necropsy
- Neuropathological findings:
- not examined
- Description (incidence and severity):
- not applicable
- Histopathological findings: non-neoplastic:
- effects observed, non-treatment-related
- Description (incidence and severity):
- The thyroid along with parathyroid was collected and preserved in 10% neutral buffer formalin for microscopic examination. The histopathology of thyroid along with parathyroid was conducted for all the dose group dams. There were no treatment related histopathological findings noticed in the any of the dose group animals during conduct of the histopathological examination in the study.
However, the observations like, hemorrhage in thyroids at G1 group (1 animal); MNC infiltration in thyroids at G2 group (1 animal); Ultimobranchial cyst at G1 group (1 animal), G2 group (3 animals) and G3 group (3 animals), Ectopic thymus at G3 group (2 animals) and G4 group (1 animal) were noted. These lesions were considered as background lesions. - Histopathological findings: neoplastic:
- not examined
- Description (incidence and severity):
- not applicable
- Other effects:
- effects observed, non-treatment-related
- Description (incidence and severity):
- The serum T3 levels (ng/mL) were 2.510, 2.572, 2.370 and 2.358 for G1 (0 mg/kg), G2 (50 mg/kg), G3 (130 mg/kg) and G4 (350 mg/kg) respectively. There were no statistically significant differences in these values and no treatment related changes for this parameter across the dose groups when compared to controls.
The serum T4 levels (ng/mL) were 66.724, 72.036, 73.684 and 79.023 for G1 (0 mg/kg), G2 (50 mg/kg), G3 (130 mg/kg) and G4 (350 mg/kg) respectively. There were no changes in serum T4 levels (ng/mL) across all the tested dose groups when compared with the vehicle control group. The values obtained across groups were comparable.
A statistically significant increase in serum T4 levels (ng/mL) in G4 group animals was observed. This incidence is considered as incidental and unrelated to treatment with test item due to comparable values across groups and the values obtained were within biological control range.
The serum TSH levels (µIU/mL) were 4.351, 4.142, 3.767 and 4.439 for G1 (0 mg/kg), G2 (50 mg/kg), G3 (130 mg/kg) and G4 (350 mg/kg) respectively. There were no statistically significant differences in serum TSH levels and no treatment related changes for this parameter across the dose groups when compared to controls. - Details on results:
- The Test item, Hordaphos MDGB was administered via oral route at a dose volume of 10 mL/kg body weight to mated female rats from gestation day 5 (GD5) through gestation day 19 (GD19). The dose levels were 0 mg/kg, 50 mg/kg, 130 mg/kg and 350 mg/kg for the control (G1), low (G2), mid (G3) and high (G4) groups, respectively. The results of the experiment support the conclusion that the NOAEL [No Observed Adverse Effect Level] for the test item, Hordaphos MDGB for maternal toxicity is 350 mg/kg, the high dose, as there were no effects noted on maternal parameters at all the tested dose groups.
Maternal developmental toxicity
- Number of abortions:
- no effects observed
- Description (incidence and severity):
- not applicable
- Pre- and post-implantation loss:
- effects observed, non-treatment-related
- Description (incidence and severity):
- The mean percentage pre-implantation loss was 0.00, 0.00, 0.00 and 0.37 for G1 (0 mg/kg), G2 (50 mg/kg), G3 (130 mg/kg) and G4 (350 mg/kg) respectively. There were no statistically significant differences in these values and no treatment related changes for this parameter across the dose groups when compared to controls.
The mean percentage of post-implantation loss was 6.70, 4.99, 9.28 and 4.64 for G1 (0 mg/kg), G2 (50 mg/kg), G3 (130 mg/kg) and G4 (350 mg/kg) respectively. There were no statistically significant differences in these values and no treatment related changes for this parameter across the dose groups when compared to controls. - Total litter losses by resorption:
- no effects observed
- Description (incidence and severity):
- not applicable
- Early or late resorptions:
- effects observed, non-treatment-related
- Description (incidence and severity):
- The mean number of early resorptions per dam was 0.57, 0.50, 0.95 and 0.43 and percentage of early resorptions was 5.71, 3.89, 8.14 and 3.25 for G1 (0 mg/kg), G2 (50 mg/kg), G3 (130 mg/kg) and G4 (350 mg/kg) respectively. There were no statistically significant differences observed in the number and percentage of early resorptions per dam across dose groups when compared to the vehicle control.
The mean number of late resorptions per dam was 0.14, 0.15, 0.14 and 0.19 and percentage of early resorptions was 1.00, 1.10, 1.13 and 1.39 for G1 (0 mg/kg), G2 (50 mg/kg), G3 (130 mg/kg) and G4 (350 mg/kg) respectively. There were no statistically significant differences observed in the number and percentage of late resorptions per dam across dose groups when compared to the vehicle control. - Dead fetuses:
- no effects observed
- Description (incidence and severity):
- There were no dead fetuses noted for any litter in any of the test dose group or the vehicle control litters
- Changes in pregnancy duration:
- no effects observed
- Description (incidence and severity):
- not applicable
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): no effects observed
Field "Description (incidence and severity)" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.DescriptionIncidenceAndSeverityEffectsOnPregnancyDuration): not applicable - Changes in number of pregnant:
- no effects observed
- Description (incidence and severity):
- not applicable
- Details on maternal toxic effects:
- No effects were noted at all the tested dose groups
Effect levels (maternal animals)
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- >= 350 mg/kg bw/day
- Based on:
- test mat.
- Basis for effect level:
- body weight and weight gain
- changes in number of pregnant
- changes in pregnancy duration
- clinical signs
- dead fetuses
- early or late resorptions
- effects on pregnancy duration
- food consumption and compound intake
- gross pathology
- histopathology: non-neoplastic
- maternal abnormalities
- mortality
- necropsy findings
- number of abortions
- organ weights and organ / body weight ratios
- pre and post implantation loss
- total litter losses by resorption
- other: Thyroid hormonal levels
Maternal abnormalities
- Key result
- Abnormalities:
- no effects observed
- Localisation:
- cervix
- ovary
- uterus
- vagina
Results (fetuses)
- Fetal body weight changes:
- effects observed, non-treatment-related
- Description (incidence and severity):
- For groups G1 (0 mg/kg), G2 (50 mg/kg), G3 (150 mg/kg) and G4 (350 mg/kg), respectively, the male mean fetal weights were 4.03 g, 4.15 g, 4.22 g and 4.14 g; the female mean fetal weights were 3.84 g, 3.85 g, 3.94 g and 3.88 g. There were no changes in mean fetal weights across all the tested dose groups when compared with the vehicle control group. The values obtained across groups were comparable.
A statistically significant increase in mean male fetal weights in G3 group was observed. This incidence is considered as incidental and unrelated to treatment with test item due to comparable values across groups and missing dose-response relationship. Also other fetal parameters are unaffected
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): effects observed, non-treatment-related
Field "Description (incidence and severity)" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.DescriptionIncidenceAndSeverityFetalPupBodyWeightChanges): For groups G1 (0 mg/kg), G2 (50 mg/kg), G3 (150 mg/kg) and G4 (350 mg/kg), respectively, the male mean fetal weights were 4.03 g, 4.15 g, 4.22 g and 4.14 g; the female mean fetal weights were 3.84 g, 3.85 g, 3.94 g and 3.88 g. There were no changes in mean fetal weights across all the tested dose groups when compared with the vehicle control group. The values obtained across groups were comparable.
A statistically significant increase in mean male fetal weights in G3 group was observed. This incidence is considered as incidental and unrelated to treatment with test item due to comparable values across groups and missing dose-response relationship. Also other fetal parameters are unaffected - Reduction in number of live offspring:
- no effects observed
- Description (incidence and severity):
- not applicable
- Changes in sex ratio:
- no effects observed
- Description (incidence and severity):
- The sex ratio calculated as the mean percentage of male fetuses was 1.36, 1.54, 1.09 and 1.19 for G1 (0 mg/kg), G2 (50 mg/kg), G3 (130 mg/kg) and G4 (350 mg/kg) respectively. There were no statistically significant differences in these values and no treatment related changes for this parameter across the dose groups when compared to controls.
- Changes in litter size and weights:
- no effects observed
- Description (incidence and severity):
- The mean litter sizes, assessed as the in utero total of fetuses [live plus dead] per dam were 12.52, 12.20, 11.95 and 12.10 for groups G1 (0 mg/kg) ,G2 (50 mg/kg), G3 (130 mg/kg) and G4 (350 mg/kg) respectively. There were no dead fetuses within any litter in any group; therefore, the numbers of live fetuses per dam across groups were the same. There were no statistically significant differences noted across dose groups when compared to the control.
- Changes in postnatal survival:
- not examined
- Description (incidence and severity):
- not applicable
- External malformations:
- effects observed, non-treatment-related
- Description (incidence and severity):
- A total of 263 (21), 244 (20), 251 (21) and 254 (21) fetuses (litters) with a mean of 12.52, 12.20, 11.95 and 12.10 fetuses per dam were available for gross external examination from groups G1 (0 mg/kg), G2 (50 mg/kg), G3 (130 mg/kg) and G4 (350 mg/kg), respectively.
No treatment related external abnormalities were noted within any group during external examination of these fetuses. However, the following variations were noted from all the group fetuses during fetal external examinations:
In G1 (vehicle control) group, haemorrhagic spot on the hind limb unilaterally in one fetus was noted.
In G3 (130 mg/kg) group, haemorrhagic spot on ventral side of trunk in one fetus was noted.
In G4 (350 mg/kg) group, haemorrhagic spot on hind limb unilaterally in one fetus, on head in one fetus and on forelimb unilaterally in one fetus were noted.
These findings are common for fetuses of this species and strain and cannot be considered as treatment related. - Skeletal malformations:
- effects observed, non-treatment-related
- Description (incidence and severity):
- The skeletal malformations reflect variations in the maturation rate of the ossified tissues of rat fetuses. They are unlikely to be detectable in juvenile animals after continued growth postnatally. These skeletal malformations did not occur in a dose-dependent pattern, nor was the severity or the incidence of the findings increased with dose.
Overall, the skeletal morphologic observations reflect variations in the maturation rate of the ossified tissues of rat fetuses; they did not occur in a dose-dependent pattern, nor was the severity of the anomaly increased with dose. These result support the conclusion that the findings are incidental and that the test item did not produce an adverse effect on the skeletal system during fetal development at any dose in this study. - Visceral malformations:
- effects observed, non-treatment-related
- Description (incidence and severity):
- The visceral observations are common findings for fetuses of this species; they did not occur in a dose-dependent pattern, nor was the severity or the incidence of the findings increased with dose. This result supports the conclusion that the findings are incidental and that the test item, Hordaphos MDGB, did not produce an adverse effect on the soft tissues during fetal development.
- Other effects:
- no effects observed
- Description (incidence and severity):
- For groups G1 (0 mg/kg), G2 (50 mg/kg), G3 (130 mg/kg) and G4 (350 mg/kg), respectively, the male fetal crown rump length was 39.02 mm, 39.77 mm, 39.90 mm and 39.77 mm; the female fetal crown rump length was 38.07 mm, 38.07 mm, 38.28 mm and 38.20 mm. There were no statistically significant differences in fetal crown rump length and no treatment related changes for this parameter across the dose groups when compared to controls.
For groups G1 (0 mg/kg), G2 (50 mg/kg), G3 (130 mg/kg) and G4 (350 mg/kg), respectively, the mean male ano-genital distance ratio was 3.02 mm, 2.98 mm, 3.02 mm and 3.06 mm; the mean female ano-genital distance ratio was 1.40 mm, 1.41 mm, 1.39 mm and 1.43 mm. There were no statistically significant differences in fetal ano-genital distance ratio and no treatment related changes for this parameter across the dose groups when compared to controls. - Details on embryotoxic / teratogenic effects:
- The Test item, Hordaphos MDGB was administered via oral route at a dose volume of 10 mL/kg body weight to mated female rats from gestation day 5 (GD5) through gestation day 19 (GD19). The dose levels were 0 mg/kg, 50 mg/kg, 130 mg/kg and 350 mg/kg for the control (G1), low (G2), mid (G3) and high (G4) groups, respectively. The results of the experiment support the conclusion that the NOAEL [No Observed Adverse Effect Level] for the test item, Hordaphos MDGB for developmental toxicity is 350 mg/kg, the high dose, as there were no effects noted on fetal parameters at all the tested dose groups.
Effect levels (fetuses)
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- ca. 350 mg/kg bw/day
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- reduction in number of live offspring
- changes in sex ratio
- fetal/pup body weight changes
- changes in litter size and weights
- external malformations
- skeletal malformations
- visceral malformations
- other: Ano-genital distance and crown rump length
Fetal abnormalities
- Key result
- Abnormalities:
- effects observed, non-treatment-related
- Localisation:
- external: cranium
- external: ear
- external: eye
- external: face
- external: limb
- external: paw
- external: tail
- external: trunk
- external: anogenital distance
- external: anus
- external: genital tubercle
- external: large intestine
- external: thorax
- external: umbilicus
- external: pelvic region
- skeletal: skull
- skeletal: skull, fontanelles
- skeletal: skull sutures
- skeletal: clavicle
- skeletal: scapule
- skeletal: forelimb
- skeletal: sternum
- skeletal: rib
- skeletal: supernumerary rib
- skeletal: vertebra
- skeletal: pelvic girdle
- skeletal: hindlimb
- visceral/soft tissue: integumentary
- visceral/soft tissue: gastrointestinal tract
- visceral/soft tissue: hepatobiliary
- visceral/soft tissue: urinary
- visceral/soft tissue: cardiovascular
- visceral/soft tissue: heamatopoietic
- visceral/soft tissue: immune system
- visceral/soft tissue: musculoskeletal system
- visceral/soft tissue: nervous system
- visceral/soft tissue: central nervous system
- visceral/soft tissue: peripheral nervous system
- visceral/soft tissue: somatic nervous system
- visceral/soft tissue: autonomic nervous system
- visceral/soft tissue: endocrine system
- visceral/soft tissue: respiratory system
- visceral/soft tissue: male reproductive system
- visceral/soft tissue: female reproductive system
- visceral/soft tissue: eye
- visceral/soft tissue: ear
- other:
Overall developmental toxicity
- Key result
- Developmental effects observed:
- no
- Lowest effective dose / conc.:
- 50 mg/kg bw/day
- Treatment related:
- no
Any other information on results incl. tables
SUMMARY OF THE STUDY
Parameters↓ |
Group |
G1 |
G2 |
G3 |
G4 |
Dose |
0 |
50 |
130 |
350 |
|
PREGNANCY DATA |
|||||
Initial Animals per Group |
25 |
25 |
25 |
25 |
|
No. of animals survived |
25 |
25 |
25 |
25 |
|
No. of animals aborted |
0 |
0 |
0 |
0 |
|
No. of animals delivering early |
0 |
0 |
0 |
0 |
|
No. of animals confirmed with pregnancy at necropsy |
21 |
20 |
21 |
21 |
|
Pregnancy rate (%) |
84.0 |
80.0 |
84.0 |
84.0 |
|
MATERNAL BODY WEIGHT |
|||||
Maternal Body Weight (g) on Gestation Day 5 |
Mean |
272.42 |
278.74 |
273.34 |
274.52 |
±SD |
16.95 |
21.58 |
14.87 |
11.68 |
|
Maternal Body Weight (g) on Gestation Day 20 |
Mean |
374.77 |
383.01 |
374.23 |
377.60 |
±SD |
24.23 |
30.70 |
25.35 |
16.48 |
|
Body Weight Change (g) during Gestation Day 5 to 20 |
Mean |
102.35 |
104.27 |
100.89 |
103.07 |
±SD |
20.71 |
24.57 |
18.23 |
13.18 |
|
Corrected Body Weight (g) |
Mean |
27.51 |
30.75 |
27.34 |
28.40 |
±SD |
15.45 |
13.97 |
13.54 |
11.74 |
|
UTERINE OBSERVATION AND MATERNAL DATA |
|||||
Gravid Uterus Weight (g) |
Mean |
74.84 |
73.52 |
73.55 |
74.67 |
±SD |
18.15 |
16.22 |
16.60 |
11.53 |
|
Corpora Lutea (no.) |
Mean |
13.24 |
12.85 |
13.05 |
12.76 |
±SD |
3.06 |
3.48 |
2.50 |
2.30 |
|
Implantation per Dam (no.) |
Mean |
13.24 |
12.85 |
13.05 |
12.71 |
±SD |
3.06 |
3.48 |
2.50 |
2.31 |
|
Male/female Sex Ratio (no.) |
Mean |
1.36 |
1.54 |
1.09 |
1.19 |
±SD |
0.95 |
1.07 |
0.91 |
0.64 |
|
Litter Size (no.) |
Mean |
12.52 |
12.20 |
11.95 |
12.10 |
±SD |
3.57 |
3.37 |
2.96 |
2.14 |
|
Live Fetuses per Dam (no.) |
Mean |
12.52 |
12.20 |
11.95 |
12.10 |
±SD |
3.57 |
3.37 |
2.96 |
2.14 |
|
Percent of Live Fetuses (%) |
Mean |
100.00 |
100.00 |
100.00 |
100.00 |
±SD |
0.00 |
0.00 |
0.00 |
0.00 |
|
Dead Fetuses per Dam (no.) |
Mean |
0.00 |
0.00 |
0.00 |
0.00 |
±SD |
0.00 |
0.00 |
0.00 |
0.00 |
|
Percent of Dead Fetuses (%) |
Mean |
0.00 |
0.00 |
0.00 |
0.00 |
±SD |
0.00 |
0.00 |
0.00 |
0.00 |
SUMMARY OF THE STUDY (Contd…).
Parameters↓ |
Group |
G1 |
G2 |
G3 |
G4 |
Dose |
0 |
50 |
130 |
350 |
|
Early Resorptions per Dam (no.) |
Mean |
0.57 |
0.50 |
0.95 |
0.43 |
±SD |
0.93 |
0.69 |
0.92 |
0.68 |
|
Percent of Early Resorptions (%) |
Mean |
5.71 |
3.89 |
8.14 |
3.25 |
±SD |
10.08 |
5.52 |
8.36 |
5.17 |
|
Late Resorptions per Dam (no.) |
Mean |
0.14 |
0.15 |
0.14 |
0.19 |
±SD |
0.48 |
0.37 |
0.36 |
0.40 |
|
Percent of Late Resorptions (%) |
Mean |
1.00 |
1.10 |
1.13 |
1.39 |
±SD |
3.37 |
2.69 |
2.85 |
2.94 |
|
Pre-implantation Loss (%) |
Mean |
0.00 |
0.00 |
0.00 |
0.37 |
±SD |
0.00 |
0.00 |
0.00 |
1.68 |
|
Post-implantation Loss (%) |
Mean |
6.70 |
4.99 |
9.28 |
4.64 |
±SD |
10.54 |
6.69 |
8.41 |
5.09 |
|
MATERNAL THYROID ALONG WITH PARATHYROID WEIGHT |
|||||
Thyroid along with parathyroid Weight (g) |
Mean |
0.0232 |
0.0220 |
0.0236 |
0.0235 |
±SD |
0.0028 |
0.0024 |
0.0019 |
0.0031 |
|
MATERNAL THYROID HORMONE LEVELS |
|||||
Serum T3 Levels (ng/mL) |
Mean |
2.510 |
2.572 |
2.370 |
2.358 |
±SD |
0.253 |
0.250 |
0.183 |
0.182 |
|
Serum T4 Levels (ng/mL) |
Mean |
66.724 |
72.036 |
73.684 |
79.023* |
±SD |
8.319 |
9.857 |
11.366 |
12.286 |
|
Serum TSH Levels (µIU/mL)) |
Mean |
4.351 |
4.142 |
3.767 |
4.439 |
±SD |
2.948 |
2.997 |
1.247 |
2.174 |
|
LITTER DATA |
|||||
Live Male Fetuses (no.) |
Mean |
6.43 |
6.60 |
5.48 |
6.24 |
±SD |
2.42 |
1.90 |
2.18 |
2.17 |
|
Live Female Fetuses (no.) |
Mean |
6.10 |
5.60 |
6.48 |
5.86 |
±SD |
2.70 |
2.64 |
2.62 |
1.65 |
|
Male Fetal Weight per Dam (g) |
Mean |
4.03 |
4.15 |
4.22* |
4.14 |
±SD |
0.25 |
0.33 |
0.21 |
0.27 |
|
Female Fetal Weight per Dam (g) |
Mean |
3.84 |
3.85 |
3.94 |
3.88 |
±SD |
0.29 |
0.35 |
0.25 |
0.30 |
|
Male Fetal Crown Rump Length per Dam (mm) |
Mean |
39.020 |
39.768 |
39.899 |
39.774 |
±SD |
1.45 |
2.00 |
1.39 |
2.75 |
|
Female Fetal Crown Rump Length per Dam (mm) |
Mean |
38.069 |
38.066 |
38.276 |
38.203 |
±SD |
1.83 |
2.44 |
1.61 |
2.38 |
|
Mean Male Ano-genital Distance Ratio |
Mean |
3.02 |
2.98 |
3.02 |
3.06 |
±SD |
0.18 |
0.19 |
0.16 |
0.16 |
|
Mean Female Ano-genital Distance Ratio |
Mean |
1.40 |
1.41 |
1.39 |
1.43 |
±SD |
0.29 |
0.17 |
0.19 |
0.15 |
SUMMARY OF THE STUDY (Contd…).
Parameters↓ |
Group |
G1 |
G2 |
G3 |
G4 |
Dose (mg/kg body weight) |
0 |
50 |
130 |
350 |
|
FETAL DATA |
|||||
External Examination |
|||||
Total No. of Fetuses for Gross External Examination |
263 |
244 |
251 |
254 |
|
Number of fetuses observed with No Abnormality Detected |
262 |
244 |
250 |
251 |
|
Number of fetuses observed with Malformations |
0 |
0 |
0 |
0 |
|
Number of fetuses observed with Variations |
1 |
0 |
1 |
3 |
|
Visceral Examination |
|||||
Total No. of Fetuses for Visceral Examination |
125 |
118 |
118 |
123 |
|
Number of fetuses observed with No Abnormality Detected |
117 |
111 |
111 |
116 |
|
Number of fetuses observed with Malformations |
0 |
1 |
1 |
0 |
|
Number of fetuses observed with Variations |
8 |
6 |
6 |
7 |
|
Skeletal Examination |
|||||
Total No. of Fetuses for Skeletal Examination |
138 |
126 |
133 |
131 |
|
Number of fetuses observed with No Abnormality Detected |
100 |
92 |
92 |
95 |
|
Number of fetuses observed with Malformations |
18 |
16 |
17 |
16 |
|
Number of fetuses observed with Variations |
20 |
18 |
24 |
20 |
TABLE 1. SUMMARY OF CLINICAL SIGNS OF TOXICITY AND MORTALITY RECORD
Group & Dose (mg/kg body weight) |
No. of Animals |
Clinical Signs of Toxicity |
Mortality |
|
G1 & 0 |
25 |
N (25) |
0/25 |
|
G2 & 50 |
25 |
N (25) |
0/25 |
|
G3 & 130 |
25 |
N (25) |
0/25 |
|
G4 & 350 |
25 |
N (25) |
0/25 |
|
N: Normal
TABLE 2. SUMMARY OF GESTATION BODY WEIGHT (g)RECORD
Group & Dose |
Body Weight (g) on Gestation Day |
|||||||||
0 |
3 |
5 |
8 |
11 |
14 |
17 |
19 |
20 |
||
G1 & 0 |
Mean |
257.63 |
264.61 |
272.42 |
279.76 |
292.08 |
307.78 |
331.61 |
358.50 |
374.77 |
±SD |
15.33 |
16.85 |
16.95 |
17.04 |
18.80 |
19.57 |
21.90 |
23.30 |
24.23 |
|
n |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
|
G2 & 50 |
Mean |
264.13 |
272.42 |
278.74 |
286.78 |
300.98 |
318.11 |
345.75 |
369.81 |
383.01 |
±SD |
16.51 |
18.48 |
21.58 |
19.70 |
21.61 |
22.68 |
28.87 |
31.76 |
30.70 |
|
n |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
|
G3 & 130 |
Mean |
258.60 |
266.10 |
273.34 |
283.31 |
296.38 |
311.77 |
336.83 |
359.24 |
374.23 |
±SD |
15.37 |
15.54 |
14.87 |
15.77 |
15.38 |
19.56 |
21.46 |
26.01 |
25.35 |
|
n |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
|
G4 & 350 |
Mean |
258.30 |
266.35 |
274.52 |
282.45 |
293.93 |
310.29 |
337.05 |
361.51 |
377.60 |
±SD |
10.06 |
9.96 |
11.68 |
14.11 |
14.47 |
15.82 |
15.29 |
17.06 |
16.48 |
|
n |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
SD: Standard Deviation; n: number of dams
Note: The data of non-pregnant females is not included in mean calculations and not subjected to statistical analysis.
TABLE 3. SUMMARY OF PERCENT CHANGE IN GESTATION BODY WEIGHT (%) RECORD
Group & Dose |
Percent (%) Change in Body Weight during Gestation Day |
||||||||||
0 to 3 |
3 to 5 |
5 to 8 |
8 to 11 |
11 to 14 |
14 to 17 |
17 to 19 |
19 to 20 |
0 to 20 |
5 to 20 |
||
G1 & 0 |
Mean |
2.70 |
2.97 |
2.71 |
4.40 |
5.40 |
7.75 |
8.15 |
4.55 |
45.73 |
37.80 |
±SD |
1.95 |
1.57 |
1.70 |
1.98 |
2.01 |
2.48 |
2.70 |
1.80 |
9.43 |
8.41 |
|
n |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
|
G2 & 50 |
Mean |
3.13 |
2.27 |
2.96 |
4.95 |
5.74 |
8.64 |
6.97 |
3.63 |
45.09 |
37.69 |
±SD |
2.14 |
1.81 |
1.91 |
1.83 |
3.45 |
3.47 |
2.53 |
1.88 |
8.90 |
9.63 |
|
n |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
|
G3 & 130 |
Mean |
2.92 |
2.75 |
3.65 |
4.65 |
5.15 |
8.06 |
6.61 |
4.22 |
44.80 |
36.96 |
±SD |
1.79 |
1.81 |
1.48 |
2.19 |
2.08 |
2.40 |
2.09 |
1.31 |
7.06 |
6.60 |
|
n |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
|
G4 & 350 |
Mean |
3.15 |
3.06 |
2.87 |
4.08 |
5.58 |
8.67 |
7.28 |
4.48 |
46.26 |
37.64 |
±SD |
2.15 |
1.94 |
1.71 |
1.53 |
1.94 |
2.18 |
2.52 |
1.64 |
5.42 |
5.41 |
|
n |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
SD: Standard Deviation; n: No. of dams
Note: The data of non-pregnant females is not included in mean calculations and not subjected to statistical analysis.
TABLE 4. SUMMARY OFGRAVID UTERUS WEIGHT (g) AND MATERNAL BODYWEIGHT CHANGE CORRECTED FOR GRAVID UTERINE WEIGHT (g) RECORD
Group & Dose (mg/kg body weight) |
Gravid Uterus Weight (g) |
Body Weight Change (g) from GD 5 to 20 |
Corrected Body weight (g) |
Corrected (%) |
||
G1 & 0 |
Mean |
74.84 |
102.35 |
27.51 |
10.30 |
|
±SD |
18.15 |
20.71 |
15.45 |
5.99 |
||
n |
21 |
21 |
21 |
21 |
||
G2 & 50 |
Mean |
73.52 |
104.27 |
30.75 |
11.18 |
|
±SD |
16.22 |
24.57 |
13.97 |
5.27 |
||
n |
20 |
20 |
20 |
20 |
||
G3 & 130 |
Mean |
73.55 |
100.89 |
27.34 |
10.03 |
|
±SD |
16.60 |
18.23 |
13.54 |
4.92 |
||
n |
21 |
21 |
21 |
21 |
||
G4 & 350 |
Mean |
74.67 |
103.07 |
28.40 |
10.42 |
|
±SD |
11.53 |
13.18 |
11.74 |
4.56 |
||
n |
21 |
21 |
21 |
21 |
SD: Standard Deviation; n: No. of dams
Note: The data of non-pregnant females is not included in mean calculations and not subjected to statistical analysis.
TABLE 5. SUMMARY OF AVERAGE GESTATION FEED CONSUMPTION (g/animal/day) RECORD
Group & Dose (mg/kg body weight) |
Feed Consumption (g/animal/day) during Gestation Day |
||||||||||
0 to 3 |
3 to 5 |
5 to 8 |
8 to 11 |
11 to 14 |
14 to 17 |
17 to 19 |
19 to 20 |
0 to 20 |
5 to 20 |
||
G1 & 0 |
Mean |
18.68 |
18.89 |
19.88 |
21.75 |
22.38 |
24.66 |
25.23 |
25.60 |
22.13 |
23.25 |
±SD |
2.84 |
3.74 |
3.84 |
3.15 |
3.93 |
4.18 |
4.96 |
8.28 |
1.94 |
2.37 |
|
n |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
|
G2 & 50 |
Mean |
18.50 |
19.69 |
19.94 |
21.97 |
22.57 |
24.02 |
29.20* |
35.08* |
23.87 |
25.46 |
±SD |
2.31 |
2.70 |
2.97 |
3.84 |
4.22 |
4.14 |
4.47 |
15.78 |
2.62 |
3.13 |
|
n |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
|
G3 & 130 |
Mean |
17.74 |
19.94 |
20.73 |
21.80 |
23.01 |
24.77 |
27.92 |
31.07 |
23.37 |
24.88 |
±SD |
2.32 |
2.72 |
3.28 |
3.32 |
3.08 |
4.05 |
3.28 |
10.98 |
1.62 |
1.88 |
|
n |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
|
G4 & 350 |
Mean |
18.90 |
20.66* |
20.22 |
21.55 |
22.68 |
23.94 |
28.69 |
32.07 |
23.59 |
24.86 |
±SD |
2.71 |
3.25 |
3.72 |
3.96 |
3.85 |
3.62 |
3.66 |
13.82 |
2.67 |
2.97 |
|
n |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
SD: Standard Deviation; n: Number of dams;*. The mean difference is significant at the 0.05 level.
Note: The data of non-pregnant females is not included in mean calculations and not subjected to statistical analysis.
TABLE 6. SUMMARY OF UTERI OBSERVATION RECORD
Group & Dose (mg/kg body weight) |
No. of Corpora lutea |
No. of Implantations |
Litter Size |
No. of Live Fetuses |
No. of Male Live Fetuses |
No. of Female Live Fetuses |
No. of Dead Fetuses |
No. of |
No. of |
|
G1 & 0 |
Mean |
13.24 |
13.24 |
12.52 |
12.52 |
6.43 |
6.10 |
0.00 |
0.57 |
0.14 |
±SD |
3.06 |
3.06 |
3.57 |
3.57 |
2.42 |
2.70 |
0.00 |
0.93 |
0.48 |
|
n |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
|
G2 & 50 |
Mean |
12.85 |
12.85 |
12.20 |
12.20 |
6.60 |
5.60 |
0.00 |
0.50 |
0.15 |
±SD |
3.48 |
3.48 |
3.37 |
3.37 |
1.90 |
2.64 |
0.00 |
0.69 |
0.37 |
|
n |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
|
G3 & 130 |
Mean |
13.05 |
13.05 |
11.95 |
11.95 |
5.48 |
6.48 |
0.00 |
0.95 |
0.14 |
±SD |
2.50 |
2.50 |
2.96 |
2.96 |
2.18 |
2.62 |
0.00 |
0.92 |
0.36 |
|
n |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
|
G4 & 350 |
Mean |
12.76 |
12.71 |
12.10 |
12.10 |
6.24 |
5.86 |
0.00 |
0.43 |
0.19 |
±SD |
2.30 |
2.31 |
2.14 |
2.14 |
2.17 |
1.65 |
0.00 |
0.68 |
0.40 |
|
n |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
SD: Standard Deviation; n: Number of dams;*. The mean difference is significant at the 0.05 level.
Note: The data of non-pregnant females is not included in mean calculations and not subjected to statistical analysis.
TABLE 7. SUMMARY OF MATERNAL DATA RECORD
Group & Dose (mg/kg body weight) |
|
Pre-Implantation Loss (%) |
Post-Implantation Loss (%) |
Percent of Dead Fetus (%) |
Percent of Early Resorptions (%) |
Percent of Late Resorptions(%) |
Male / Female Sex ratio |
Male Fetuses (%) |
Female Fetuses (%) |
Percent of Live Fetuses (%) |
|
G1 & 0 |
Mean |
0.00 |
6.70 |
0.00 |
5.71 |
1.00 |
1.36 |
52.15 |
47.85 |
100.00 |
|
±SD |
0.00 |
10.54 |
0.00 |
10.08 |
3.37 |
0.95 |
15.35 |
15.35 |
0.00 |
||
n |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
||
G2 & 50 |
Mean |
0.00 |
4.99 |
0.00 |
3.89 |
1.10 |
1.54 |
55.71 |
44.29 |
100.00 |
|
±SD |
0.00 |
6.69 |
0.00 |
5.52 |
2.69 |
1.07 |
13.67 |
13.67 |
0.00 |
||
n |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
||
G3 & 130 |
Mean |
0.00 |
9.28 |
0.00 |
8.14 |
1.13 |
1.09 |
45.81 |
54.19 |
100.00 |
|
±SD |
0.00 |
8.41 |
0.00 |
8.36 |
2.85 |
0.91 |
16.45 |
16.45 |
0.00 |
||
n |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
||
G4 & 350 |
Mean |
0.37 |
4.64 |
0.00 |
3.25 |
1.39 |
1.19 |
51.52 |
48.72 |
100.00 |
|
±SD |
1.68 |
5.09 |
0.00 |
5.17 |
2.94 |
0.64 |
15.33 |
11.66 |
0.00 |
||
n |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
21 |
SD: Standard Deviation; n: Number of dams
Sex Ratio Calculation:
Male/Female Sex Ratio = Total Number of Live Male Fetuses / Total Number of Live Female Fetuses
Percent of Live Fetuses (%):
Percent of Male Fetuses (%) = Total Number of live male fetuses / Total number of live fetuses x 100
Percent of Female Fetuses (%) = Total Number of live female fetuses / Total number of live fetuses x 100
TABLE 8. SUMMARY OFABSOLUTE ORGAN WEIGHT (g) AND ORGAN WEIGHT RELATIVE TO TERMINAL BODY WEIGHT (%) RECORD
Group & Dose |
|
Absolute Weight of Thyroid along with parathyroid# |
Terminal Body Weight (g) |
Relative Thyroid along with parathyroid Weight (%) |
G1 & 0 |
Mean |
0.0232 |
374.77 |
0.0062 |
±SD |
0.0028 |
24.23 |
0.0008 |
|
n |
21 |
21 |
21 |
|
G2 & 50 |
Mean |
0.0220 |
383.01 |
0.0058 |
±SD |
0.0024 |
30.70 |
0.0008 |
|
n |
20 |
20 |
20 |
|
G3 & 130 |
Mean |
0.0236 |
374.23 |
0.0063 |
±SD |
0.0019 |
25.35 |
0.0006 |
|
n |
21 |
21 |
21 |
|
G4 & 350 |
Mean |
0.0235 |
377.60 |
0.0062 |
±SD |
0.0031 |
16.48 |
0.0009 |
|
n |
21 |
21 |
21 |
SD: Standard Deviation; n: Number of dams; #: Weighed post fixation
TABLE 9. SUMMARY OF SERUM TRIIODOTHYRONINE (T3) LEVELS (ng/mL) RECORD
Group & Dose |
|
Serum T3 Levels (ng/mL) |
|
G1 & 0 |
Mean |
2.510 |
|
±SD |
0.253 |
||
n |
21 |
||
G2 & 50 |
Mean |
2.572 |
|
±SD |
0.250 |
||
n |
20 |
||
G3 & 130 |
Mean |
2.370 |
|
±SD |
0.183 |
||
n |
21 |
||
G4 & 350 |
Mean |
2.358 |
|
±SD |
0.182 |
||
n |
21 |
SD: Standard Deviation; n: Number of dams
TABLE 10. SUMMARY OF SERUM THYROXINE (T4) LEVELS (ng/mL) RECORD
Group & Dose |
|
Serum T4 Levels (ng/mL) |
|
G1 & 0 |
Mean |
66.724 |
|
±SD |
8.319 |
||
n |
21 |
||
G2 & 50 |
Mean |
72.036 |
|
±SD |
9.857 |
||
n |
20 |
||
G3 & 130 |
Mean |
73.684 |
|
±SD |
11.366 |
||
n |
21 |
||
G4 & 350 |
Mean |
79.023* |
|
±SD |
12.286 |
||
n |
21 |
SD: Standard Deviation; n: Number of dams
*. The mean difference is significant at the 0.05 level.
TABLE 11. SUMMARY OF SERUM THYROID STIMULATING HORMONE (TSH) LEVELS (µIU/mL) RECORD
Group & Dose |
|
Serum TSH Levels (µIU/mL) |
|
G1 & 0 |
Mean |
4.351 |
|
±SD |
2.948 |
||
n |
21 |
||
G2 & 50 |
Mean |
4.142 |
|
±SD |
2.997 |
||
n |
20 |
||
G3 & 130 |
Mean |
3.767 |
|
±SD |
1.247 |
||
n |
21 |
||
G4 & 350 |
Mean |
4.439 |
|
±SD |
2.174 |
||
n |
21 |
SD: Standard Deviation; n: Number of dams
TABLE 12. SUMMARY OF MEAN FETAL WEIGHT (g) RECORD
SD: Standard Deviation; n: Number of dams *. The mean difference is significant at the 0.05 level.
TABLE 13. SUMMARY OF MEAN FETAL CROWN RUMP LENGTH (mm) RECORD
SD: Standard Deviation; n: Number of dams
TABLE 14. SUMMARY OF MEAN FETAL ANOGENITAL DISTANCE (AGD) RATIO RECORD
SD: Standard Deviation; n: Number of dams
|
TABLE 15. SUMMARY OF EXTERNAL EXAMINATION OF FETUSES
Parameters |
Group |
G1 |
G2 |
G3 |
G4 |
Dose (mg/kg body weight) |
0 |
50 |
130 |
350 |
|
Number of Dams |
21 |
20 |
21 |
21 |
|
Number of fetuses for External Examination |
263 |
244 |
251 |
254 |
|
Number of fetuses observed with No Abnormality Detected |
262 |
244 |
250 |
251 |
|
Number of fetuses observed with Malformations |
0 |
0 |
0 |
0 |
|
Number of fetuses observed with Variations |
1 |
0 |
1 |
3 |
|
VARIATIONS |
|||||
Haemorrhagic spot - Hind limb unilateral |
No. |
1 (1) |
0 (0) |
0 (0) |
1 (1) |
% |
0.38 |
0.00 |
0.00 |
0.39 |
|
Haemorrhagic spot - Ventral side of trunk |
No. |
0 (0) |
0 (0) |
1 (1) |
0 (0) |
% |
0.00 |
0.00 |
0.40 |
0.00 |
|
Haemorrhagic spot – head region |
No. |
0 (0) |
0 (0) |
0 (0) |
1 (1) |
% |
0.00 |
0.00 |
0.00 |
0.39 |
|
Haemorrhagic spot - |
No. |
0 (0) |
0 (0) |
0 (0) |
1 (1) |
% |
0.00 |
0.00 |
0.00 |
0.39 |
Number of dams in parentheses
|
|
No. of fetuses with abnormality |
% of Abnormality |
: |
------------------------------------------ X 100 |
|
|
Total No. of fetuses examined |
TABLE 16. SUMMARY RECORD OF VISCERAL EXAMINATION OF FETUSES
Parameters |
Group |
G1 |
G2 |
G3 |
G4 |
Dose (mg/kg body weight) |
0 |
50 |
130 |
350 |
|
Number of Dams |
21 |
20 |
21 |
21 |
|
Number of fetuses for visceral examination |
125 |
118 |
118 |
123 |
|
Number of fetuses observed with No Abnormality Detected |
117 |
111 |
111 |
116 |
|
Number of fetuses observed with Malformations |
0 |
1 |
1 |
0 |
|
Number of fetuses observed with Variations |
8 |
6 |
6 |
7 |
|
MALFORMATIONS |
|||||
|
|||||
Liver Lobes Fused |
No. |
0 (0) |
0 (0) |
1 (1) |
0 (0) |
% |
0.00 |
0.00 |
0.85 |
0.00 |
|
Adrenals Smaller (unilateral) |
No. |
0 (0) |
1 (1) |
0 (0) |
0 (0) |
% |
0.00 |
0.85 |
0.00 |
0.00 |
|
VARIATIONS |
|||||
Kidney - Renal pelvis Dilation – Unilateral (right) |
No. |
2(2) |
1(1) |
1(1) |
2(2) |
% |
1.60 |
0.85 |
0.85 |
1.63 |
|
Kidney – Pale colored |
No. |
0 (0) |
0 (0) |
0 (0) |
1(1) |
% |
0.00 |
0.00 |
0.00 |
0.81 |
|
Kidney – Malpositioned |
No. |
4(4) |
4(3) |
4(3) |
3(3) |
% |
3.20 |
3.39 |
3.39 |
2.44 |
|
Microopthalmia |
No. |
2(2) |
1(1) |
0 (0) |
0 (0) |
% |
1.60 |
0.85 |
0.00 |
0.00 |
Number of dams in parentheses
|
|
No. of fetuses with abnormality |
% of Abnormality |
: |
----------------------------------------- X 100 |
|
|
Total No. of fetuses examined |
TABLE 17. SUMMARY RECORD OF SKELETAL EXAMINATION OF FETUSES
Parameters |
Group |
G1 |
G2 |
G3 |
G4 |
Dose (mg/kg body weight) |
0 |
50 |
130 |
350 |
|
Number of Dams |
21 |
20 |
21 |
21 |
|
Number of fetuses for skeletal examination |
138 |
126 |
133 |
131 |
|
Number of fetuses observed with No Abnormality Detected |
100 |
92 |
92 |
95 |
|
Number of fetuses observed with Malformations |
18 |
16 |
17 |
16 |
|
MALFORMATIONS |
|||||
SKULL |
|||||
Skull – Interparietal - Misshapen |
No. |
0(0) |
0(0) |
1(1) |
1(1) |
% |
0.00 |
0.00 |
0.75 |
0.76 |
|
STERNUM |
|||||
Sternum No. 5 - Absent |
No. |
3(3) |
4(2) |
3(3) |
5(5) |
% |
2.17 |
3.17 |
2.26 |
3.82 |
|
Sternum No. 5 & 6 - Absent |
No. |
3(3) |
0(0) |
0(0) |
0(0) |
% |
2.17 |
0.00 |
0.00 |
0.00 |
|
RIBS |
|||||
Rib No.13 short (unilateral) |
No. |
1(1) |
0(0) |
0(0) |
1(1) |
% |
0.72 |
0.00 |
0.00 |
0.76 |
|
THORACIC VERTEBRAE |
|||||
Centra No.10 Split |
No. |
0(0) |
0(0) |
0(0) |
1(1) |
% |
0.00 |
0.00 |
0.00 |
0.76 |
|
Centra No.11 Split |
No. |
0(0) |
1(1) |
0(0) |
2(2) |
% |
0.00 |
0.79 |
0.00 |
1.53 |
|
Centra No.12 Split |
No. |
1(1) |
1(1) |
2(2) |
1(1) |
% |
0.72 |
0.79 |
1.50 |
0.76 |
|
Centra No.13 Split |
No. |
1(1) |
3(2) |
0(0) |
0(0) |
% |
0.72 |
2.38 |
0.00 |
0.00 |
|
Centra No.11 & 12 Split |
No. |
0(0) |
0(0) |
1(1) |
0(0) |
% |
0.00 |
0.00 |
0.75 |
0.00 |
|
LUMBAR VERTEBRAE |
|||||
Centra No. 1 & 2 |
No. |
1(1) |
0(0) |
0(0) |
0(0) |
% |
0.72 |
0.00 |
0.00 |
0.00 |
|
FORE LIMB |
|||||
Metacarpal No.5 Absent (Bilateral) |
No. |
2(2) |
2(2) |
1(1) |
1(1) |
% |
1.45 |
1.59 |
0.75 |
0.76 |
|
Proximal phalanges No. 3 and 4 - Absent (Bilateral) |
No. |
6(4) |
5(5) |
9(9) |
4(4) |
% |
4.35 |
3.97 |
6.77 |
3.05 |
Number of dams in parentheses
|
|
No. of fetuses with abnormality |
% of Abnormality |
: |
------------------------------------------ X 100 |
|
|
Total No. of fetuses examined |
TABLE 17 (Contd...). SUMMARY OF SKELETAL EXAMINATION OF FETUSES
Parameters |
Group |
G1 |
G2 |
G3 |
G4 |
Dose (mg/kg body weight) |
0 |
50 |
130 |
350 |
|
Number of Dams |
21 |
20 |
21 |
21 |
|
Number of fetuses for skeletal examination |
138 |
126 |
133 |
131 |
|
Number of fetuses observed with |
100 |
92 |
92 |
95 |
|
Number of fetuses observed withVariations |
20 |
18 |
24 |
20 |
|
VARIATIONS |
|||||
SKULL |
|||||
Intra parietal bones - Poor Ossification |
No. |
1(1) |
0(0) |
0(0) |
0(0) |
% |
0.72 |
0.00 |
0.00 |
0.00 |
|
Intra parietal bones - Incomplete Ossification |
No. |
0(0) |
0(0) |
1(1) |
0(0) |
% |
0.00 |
0.00 |
0.75 |
0.00 |
|
Supraoccipital bones - Poor Ossification |
No. |
0(0) |
1(1) |
0(0) |
0(0) |
% |
0.00 |
0.79 |
0.00 |
0.00 |
|
Intra parietal bones & Supraoccipital bones - Incomplete Ossification |
No. |
0(0) |
0(0) |
0(0) |
1(1) |
% |
0.00 |
0.00 |
0.00 |
0.76 |
|
STERNUM |
|||||
Sternum No. 5 - Poor Ossification |
No. |
1(1) |
0(0) |
0(0) |
0(0) |
% |
0.72 |
0.00 |
0.00 |
0.00 |
|
Sternum No. 5 - Incomplete Ossification |
No. |
1(1) |
1(1) |
4(3) |
8(8) |
% |
0.72 |
0.79 |
3.01 |
6.11 |
|
Sternum No. 5 - Bipartite Ossification |
No. |
0(0) |
0(0) |
2(2) |
0(0) |
% |
0.00 |
0.00 |
1.50 |
0.00 |
|
Sternum No. 6 - Poor Ossification |
No. |
4(3) |
6(4) |
2(2) |
0(0) |
% |
2.90 |
4.76 |
1.50 |
0.00 |
|
Sternum No. 6 - Incomplete Ossification |
No. |
0(0) |
0(0) |
4(4) |
0(0) |
% |
0.00 |
0.00 |
3.00 |
0.00 |
|
Sternum No. 5 and 6 - Poor Ossification |
No. |
1(1) |
1(1) |
3(3) |
0(0) |
% |
0.72 |
0.79 |
2.26 |
0.00 |
|
Number of dams in parentheses |
|
|
No. of fetuses with abnormality |
% of Abnormality |
: |
------------------------------------------ X 100 |
|
|
Total No. of fetuses examined |
TABLE 17 (Contd…). SUMMARY OF SKELETAL EXAMINATION OF FETUSES
Parameters |
Group |
G1 |
G2 |
G3 |
G4 |
Dose (mg/kg body weight) |
0 |
50 |
130 |
350 |
|
Number of Dams |
21 |
20 |
21 |
21 |
|
Number of fetuses for skeletal examination |
138 |
126 |
133 |
131 |
|
Number of fetuses observed with |
100 |
92 |
92 |
95 |
|
Number of fetuses observed withVariations |
20 |
18 |
24 |
20 |
|
VARIATIONS |
|||||
Sternum No. 5 and 6 - Incomplete Ossification |
No. |
0(0) |
1(1) |
2(2) |
0(0) |
% |
0.00 |
0.79 |
1.50 |
0.00 |
|
Sternum No. 4 - Misshapened |
No. |
0(0) |
3(3) |
1(1) |
0(0) |
% |
0.00 |
2.38 |
0.75 |
0.00 |
|
Sternum No. 5 - Misshapened |
No. |
0(0) |
2(2) |
2(2) |
1(1) |
% |
0.00 |
1.59 |
1.50 |
0.76 |
|
RIBS |
|||||
Rib No. 13 - Wavy |
No. |
2(1) |
0(0) |
0(0) |
0(0) |
% |
1.45 |
0.00 |
0.00 |
0.00 |
|
Rib No.12 and 13 – Wavy (bilateral) |
No. |
1(1) |
0(0) |
0(0) |
0(0) |
% |
0.72 |
0.00 |
0.00 |
0.00 |
|
Rib No.11,12 and 13 – Wavy (bilateral) |
No. |
0(0) |
0(0) |
1(1) |
0(0) |
% |
0.00 |
0.00 |
0.75 |
0.00 |
|
Rib No.11 and 12 – Wavy (unilateral) |
No. |
0(0) |
0(0) |
0(0) |
1(1) |
% |
0.00 |
0.00 |
0.00 |
0.76 |
|
Rudimentary Rib |
No. |
0(0) |
0(0) |
0(0) |
2(1) |
% |
0.00 |
0.00 |
0.00 |
1.53 |
|
Rib No.10 – Wavy (unilateral) |
No. |
0(0) |
0(0) |
0(0) |
1(1) |
% |
0.00 |
0.00 |
0.00 |
0.76 |
|
THORACIC VERTEBRAE |
|||||
Centrum No. 10 – Dumbbell shaped |
No. |
0(0) |
0(0) |
0(0) |
1(1) |
% |
0.00 |
0.00 |
0.00 |
0.76 |
|
Centrum No. 11 – Assymetrical bilobed |
No. |
1(1) |
0(0) |
0(0) |
0(0) |
% |
0.72 |
0.00 |
0.00 |
0.00 |
Number of dams in parentheses
|
|
No. of fetuses with abnormality |
% of Abnormality |
: |
------------------------------------------ X 100 |
|
|
Total No. of fetuses examined |
TABLE 17 (Contd...). SUMMARY OF SKELETAL EXAMINATION OF FETUSES
Parameters |
Group |
G1 |
G2 |
G3 |
G4 |
Dose (mg/kg body weight) |
0 |
50 |
130 |
350 |
|
Number of Dams |
21 |
20 |
21 |
21 |
|
Number of fetuses for skeletal examination |
138 |
126 |
133 |
131 |
|
Number of fetuses observed with |
100 |
92 |
92 |
95 |
|
No Abnormality Detected |
|||||
Number of fetuses observed with Variations |
20 |
18 |
24 |
20 |
|
VARIATIONS |
|||||
Centrum No. 11 - Dumbbell shaped |
No. |
1(1) |
0(0) |
0(0) |
0(0) |
% |
0.72 |
0 |
0 |
0 |
|
Centrum No. 12 - Dumbbell shaped |
No. |
0(0) |
2(2) |
2(2) |
0(0) |
% |
0 |
1.59 |
1.5 |
0 |
|
Centrum No. 11 & 13 - Dumbbell shaped |
No. |
0(0) |
0(0) |
0(0) |
1(1) |
% |
0 |
0 |
0 |
0.76 |
|
Centrum No. 10, 11, 12 & 13 - Dumbbell shaped |
No. |
0(0) |
0(0) |
0(0) |
1(1) |
% |
0 |
0 |
0 |
0.76 |
|
Centrum No. 10 & 12 - Dumbbell shaped |
No. |
0(0) |
0(0) |
0(0) |
1(1) |
% |
0 |
0 |
0 |
0.76 |
|
LUMBAR VERTEBRAE |
|||||
Arch No. 4 & 5 - Incomplete Ossification |
No. |
0(0) |
1 (1) |
0(0) |
0(0) |
% |
0 |
0.79 |
0 |
0 |
|
Arch No. 5 - Incomplete Ossification |
No. |
1 (1) |
0(0) |
0(0) |
0(0) |
% |
0.79 |
0 |
0 |
0 |
|
Arch No. 1: Extra ossification site (Bilateral) |
No. |
1 (1) |
0(0) |
0(0) |
0(0) |
% |
0.79 |
0 |
0 |
0 |
|
Arch No.2: Extra Ossification Site (Unilateral) |
No. |
0(0) |
0(0) |
0(0) |
1(1) |
% |
0 |
0 |
0 |
0.76 |
|
SACRAL VERTEBRAE |
|||||
Arch No.4 : Extra Ossification Site – (Bilateral) |
No. |
2(2) |
0(0) |
0(0) |
0(0) |
% |
1.45 |
0 |
0 |
0 |
Number of dams in parentheses
|
|
No. of fetuses with abnormality |
% of Abnormality |
: |
------------------------------------------ X 100 |
|
|
Total No. of fetuses examined |
TABLE 17 (Contd...). SUMMARY OF SKELETAL EXAMINATION OF FETUSES
Parameters
|
Group |
G1 |
G2 |
G3 |
G4 |
Dose (mg/kg body weight) |
0 |
50 |
130 |
350 |
|
Number of Dams |
21 |
20 |
21 |
21 |
|
Number of fetuses for skeletal examination |
138 |
126 |
133 |
131 |
|
Number of fetuses observed with No Abnormality Detected |
100 |
92 |
92 |
95 |
|
Number of fetuses observed with Variations |
20 |
18 |
24 |
20 |
|
VARIATIONS |
|||||
CAUDAL VERTEBRAE |
|||||
Arch No.2: Incomplete Ossification |
No. |
2 (2) |
0 (0) |
0 (0) |
0 (0) |
% |
1.45 |
0 |
0 |
0 |
|
Arch No.1 & 2: Incomplete Ossification |
No. |
0 (0) |
0 (0) |
0 (0) |
1 (1) |
% |
0 |
0 |
0 |
0.76 |
|
FORE LIMB |
|||||
Proximal Phalanx No. 3 & 4 – Incomplete Ossification |
No. |
1 (1) |
0 (0) |
0 (0) |
0 (0) |
% |
0.72 |
0 |
0 |
0 |
Number of dams in parentheses
|
|
No. of fetuses with abnormality |
% of Abnormality |
: |
------------------------------------------ X 100 |
|
|
Total No. of fetuses examined |
Applicant's summary and conclusion
- Conclusions:
- The Test item, Hordaphos MDGB was administered via oral route at a dose volume of 10 mL/kg body weight to mated female rats from gestation day 5 (GD5) through gestation day 19 (GD19). The dose levels were 0 mg/kg, 50 mg/kg, 130 mg/kg and 350 mg/kg for the control (G1), low (G2), mid (G3) and high (G4) groups, respectively. The results of the experiment support the conclusion that the NOAEL [No Observed Adverse Effect Level] for the test item, Hordaphos MDGB for maternal and developmental toxicity is 350 mg/kg, the high dose, as there were no effects noted on maternal parameters as well as fetal parameters at all the tested dose groups.
- Executive summary:
The objective of this study was to assess the effects of prenatal exposure of Hordaphos MDGB on pregnant rats and on the developing fetus, including assessment of maternal effects as well as death, structural abnormalities, or altered growth in the fetus.
The test item,Hordaphos MDGB, was administered by the oral route at 10 mL/kg body weight to mated and presumed-pregnant females from Gestation Day [GD] 5 to 19. For dams the end points of assessment were maternal death, maternal body weight and clinical signs of maternal toxicity. In the fetuses the end points of assessment were fetal death, structural variations and malformations or altered growth.
A total of 100 mated female Sprague Dawley rats were allocated to four groups. Each group consisted of 25 mated presumed-pregnant female Sprague Dawley rats (Dams).Hordaphos MDGBwas administered by the oral route in graduated doses at three dose levels:
Group G1: 0 mg/kg (Vehicle Control)
Group G2: 50 mg/kg (Low Dose)
Group G3: 130 mg/kg (Mid Dose)
Group G4: 350 mg/kg (High Dose)
All the animals were observed for clinical signs of toxicity once daily, mortalities twice daily during the experimental period, the body weight and feed consumption were recorded on 0, 3, 5, 8, 11, 14, 17, 19 and on 20 (day of caesarean section). On the day of caesarean section, all the dams were observed for gravid uterus weight, uteri observations (no. of corpora lutea, no. of implantations, no. of resorptions and no. of live and foetuses). The sex ratio, pre/post implantation loss per dam, percentage of male and female foetuses was calculated based on the uteri observations. The weight of thyroid along with the parathyroid was recorded post-fixation for all the group animals.
All the animals were euthanized on gestation day 20 by exposing to CO2and subjected to detailed gross pathological examination. The histopathology of thyroid along with parathyroid conducted for all the tested dose group animals.
A total number of 21, 20, 21 and 21 mated females were confirmed pregnant across groups yielding pregnancy rates of 84%, 80%, 84% and 84% at the time of caesarean section for the G1 (0 mg/kg), G2 (50 mg/kg), G3 (130 mg/kg) and G4 (350 mg/kg), respectively.
Dose formulation analysis for dose concentration verification was performed during first week and last week of the treatment. The analysis results were within the limits and formulations were considered acceptable, as the mean results were within the range of 85 to 115% of the nominal concentration and the relative standard deviation (% RSD) is <10%.
No clinical signs of toxicity and no mortalities were noted during the experimental period at all the tested dose group animals. No treatment related effects were observed on gestational body weight, body weight gain and corrected body weight across the dose groups when compared with controls. There were no treatment related differences in feed consumption across the dose groups during gestation period.
No treatment related changes were noted in gravid uterus weight, uteri observations (no. of corpora lutea, no. of implantations, no. of resorptions and no of live and foetuses), sex ratio, pre/post implantation loss per dam, percentage of male and female foetuses and weight of thyroid along with the parathyroid wat all the tested dose groups.
No gross pathological findings were reported in all the tested dose groups during conductance of the necropsy.
The histopathology of thyroid along with parathyroid conducted for all the dose group dams did not reveal any treatment related histopathological findings during conduct of the histopathological examination. Presence ofhemorrhage in thyroid of one animal from G1 group, MNC infiltration in thyroid of one animal from G2, ultimobranchial cyst in one animal from G1 and 3 animals each from G2 and G3, ectopic thymus in 2 animals from G3 and 1 animal from G4 were considered as background lesions.
For the maternal toxicity assessment, the results of the experiment support that there were no treatment related effects on maternal body weight, feed consumption, for the parameters which assess gestational integrity end points, e.g. mean gravid uterus weight, number of corpora lutea, number of implantations, litter size, number of live fetuses, number of dead fetuses, mean number of early resorptions per dam and percentage of implantation losses per dam across all the groups.
The gravid uterus was collected by hysterectomy and fetuses were removed by caesarean section. The fetuses were observed for fetal weights, crown rump length, ano-genital distance, observation of reproductive tract, comparison of external fetal sex with internal gonads. Examination of fetuses for external, soft tissue, and skeletal anomalies was performed.
No treatment related effects on fetal weight, crown-rump length, fetal ano-genital distance ratio and comparison of external fetal sex with internal gonads were noted at all the tested dose groups.
No treatment related gross external abnormalities were noted within any group after external examination of fetuses. The noted findings of haemorrhagic spots are common findings for fetuses of this species and strain.
No treatment-related abnormalities were observed during visceral examinations of fetuses at any dose. The noted findings of renal pelvic dilation, pale colored kidneys, malposition kidneys, fused liver lobes and smaller adrenal size unilateral are anatomical variations which are common findings for fetuses of this species and strain. The observations were not dose dependent, nor were the severity of the anomaly increased with dose. This result supports the conclusion that the findings are incidental and that the test item did not produce an adverse effect on the soft tissues during fetal development.
No treatment-related abnormalities were observed during skeletal examinations of fetuses at any dose. Misshapen interparietal bones of skull, absence of sternum no. 5 and 6, short ribs, Split thoracic/lumbar vertebral centrum; absence of proximal phalanges no. 3 and 4 and metacarpal bone no. 5 were recorded as malformations. Poor/incomplete ossification ofIntra parietal/supraoccipital bones of skull,Poor/incomplete ossification and misshapen of sternum no. 5 and 6, wavy ribs, dumbbell shaped thoracic vertebrae, extra ossification site/incomplete ossification of lumbar/sacral vertebrae centrum/arch, incomplete ossification of caudal vertebrae centrum/arch and incomplete ossification of proximal phalanges no. 3 and 4 were recorded as variations. The noted anomalies are common findings for fetuses of this species and strain. They did not occur in a dose-dependent pattern, nor was the severity of the anomalies increased with dose. This result supports the conclusion that the findings are incidental and that the test item, did not produce an adverse effect on the skeletal tissues during fetal development.
For the developmental toxicity assessment, the results of the experiment support that there were no treatment related effects on the parameters which assess fetal development, i.e. fetal weight, crown-rump length, fetal ano-genital distance ratio and incidences of external, soft tissue or skeletal anomalies.
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