Registration Dossier

Diss Factsheets

Administrative data

Key value for chemical safety assessment

Toxic effect type:
dose-dependent

Effects on fertility

Description of key information

Reproductive Toxicity Study:

In a Repeated Dose Oral Toxicity Study in combination with Reproduction / Developmental Toxicity study, Wistar male and female rat treated with test chemical in the concentration of 0, 750, 1000 and 1250 mg/kg bw orally by gavage for more than 63 days. No mortality or morbidity was observed in any of the groups of animals throughout the study period however, one male was found dead on Day 42 at 750 mg/kg body weight and one male was found dead on Day 21 at 1250 mg/kg body weight after dosing due to gavaging error. No apparent treatment related clinical signs were observed in any of the animals throughout the treatment period. However, statistically significant increase was noted in urine pool at 750 mg/kg bw in male on week 2 as compared to control group. In recovery female, statistically significant decrease was noted in rearing at 1250 mg/kg bw on pre treatment, and increase in fecal bolus on week 1 as compared to the control recovery. Detailed clinical examinations like Home cage observation, Handling observation and Open field observation of all animals were observed to be normal during study period in Main study. The above changes observed were inconsistent/ biologically insignificant and not dose dependant, hence considered as incidental and not attributed to the effect of test item administration. No treatment related changes were noted in body weight and body weight gain upto 1250 mg/kg body weight. No treatment related changes were noted upto 1250 mg/kg body weight. However, Feed consumption in animals of the all other test groups of both the sexes was comparable and did not show any significant difference as compared to the respective control group. Changes observed in feed consumption were inconsistent, hence not considered as effect of the test item administration. Similarly, no treatment related changes were noted in hematological parameters upto 1250 mg/kg body weight. There were some observed variations in aPTT, PT, RBC, and platelets which were considered to be of no toxicological significance, of a minimal in nature and occurred in the absence of clear dose related response. No treatment related charges were noted in clinical chemistry parameters upto 1250 mg/kg body weight. The observed variations in clinical chemistry parameters were considered to be of no toxicological significance, of a minimal in nature and occurred in the absence of clear dose related response. No treatment related changes were noted in hormonal analysis (T4, TSH, Testosterone and Estrogen) in any of the groups. The sensory reactivity measurements were comparable and no changes were revealed in any of the animals of all treated groups in both the sexes. Foot splay and fore limb and hind limb grip strength parameters were comparable and no treatment related changes were revealed in any of the animals of all treated groups. In addition, At the end of treatment and recovery period, absolute and relative weight of organs of treated group rats of either sex did not differ significantly when compared to respective control group.  In control group G1 and treatment group G2, G3 and G4 all the females showed regular cyclicity i.e. 3-5 days estrous cycle. In cohabitation or mating period, all females from control and treated groups showed evidence of copulation i.e. sperm positive vaginal smear. All females showed precoital interval less than 5 days, except 4 females at 75 mg/kg bw which showed precoital interval more than 5 days. No differences in mating index, pre and post-implantation losses, resorption index, and sexual functions of the male and female animals were observed at any given doses. In fetal parameters,No statistically significant effect on No. of live births were observed in treated rats as compared to control. No statistically significant effect on pups weight at birth and PND13. Terminally sacrificed pups of all treated groups did not reveal any lesion of pathological significance in any of the group when compared with control group. Pups died during course of study revealed various lesions among the control and treated groups viz., external examination Cannibalism (Male: G2:2/20, Female: G2:2/24); and internal examination White Patch on edge of Liver (Male: G2: 1/18).Therefore, NOAEL was considered to be 1250 mg/kg bw for P and F1 generation Wistar male and female rat treated with test chemical orally by gavage for more than 63 days.

Link to relevant study records
Reference
Endpoint:
screening for reproductive / developmental toxicity
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Justification for type of information:
Weight of evidence based on the available data of the read-across chemicals.
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Principles of method if other than guideline:
According to OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
GLP compliance:
yes
Species:
other: Study 2 and 3: rat
Strain:
other: Study 2 and 3: Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
Study 2:
- Source: Animals were procured from a CPCSEA approved Vendor [National Institute of Biosciences, Pune Health Status:Healthy young adult animals were used for the study. Females were nulliparous and non-pregnant.
- Age: 12 - 13 weeks at the start of estrous cycle evaluation.
- Acclimatisation: Animals were acclimatised to the test conditions for 7 days prior to test item administration.Animal IdentificationDuring acclimatisation period, all the animals were identified by temporary tail marking with indelible ink and cage cards. Following allocation to the study, each animal was uniquely identified by micro toe pad tattooing and colour coded cage cards labelled with study no., study type, test system, sex, dose, group, animal number, dosing start date, experimental start and completion date. Pups were identified with body marking with permanent non-toxic marker pen.
- Husbandry:
-Housing: Before the animals are brought in, the study room and cages were cleaned and disinfected. During the study, the floor of the experimental room and work tops were swept and mopped with disinfectant solution every day or as on requirement. Cages were cleaned at regular intervals. A total 2-3 rats/sex were housed in Polycarbonate cages (size 37 [cm] x 21 [cm], height 20 [cm]). Cage rotation was carried out weekly during study period except during mating and during gestation and lactation only for females. Pregnant females were caged individually. Sterilized corn cob produced from pure corn, dried and free from dust, procured from approved supplier, was used as bedding material. It was renewed as often as necessary to keep the animals dry and clean.

- Environmental conditions: The room temperature was maintained at 19.60 to 24.70°C the relative humidity was kept between 41.80 to 66.60 %. Artificial light was set to give a cycle of 12 hours light and 12 hours dark. Air changes were about minimum 12 times per hour and filtered adequately.
- Diet A conventional laboratory pelleted diet of batch no. 040817, 041017, 040917 and 040617 from approved supplier (Nutrivet Life Sciences, Pune) was offered ad libitum.
Route of administration:
oral: gavage
Vehicle:
other: Study 2: corn oil; Study 3: Distilled water
Details on exposure:
Study 2:
PREPARATION OF DOSING SOLUTIONS:The test item was weighed and dissolved in a vehicle (corn oil) to achieve desired concentration of test item. Dose formulation was freshly prepared daily. At the time of dosing, dose formulation was kept on the magnetic stirrer to maintain the homogeneity of test item. The details of dose formulation preparation is maintained in raw data.

DIET PREPARATION
- Rate of preparation of diet (frequency): No Data Available
- Mixing appropriate amounts with (Type of food): No Data Available
- Storage temperature of food: No Data Available

VEHICLE
- Justification for use and choice of vehicle (if other than water): corn oil
- Concentration in vehicle: 0, 750, 1000 and 1250 mg/kg bw
- Amount of vehicle (if gavage): 2 mL/100g body weight
- Lot/batch no. (if required): A1708001 and A1703001
- Purity: No Data Available

Study 3:
PREPARATION OF DOSING SOLUTIONS: No data available

DIET PREPARATION
- Rate of preparation of diet (frequency): No data available
- Mixing appropriate amounts with (Type of food): No data available
- Storage temperature of food: No data available

VEHICLE
- Justification for use and choice of vehicle (if other than water): No data available
- Concentration in vehicle: 1000 mg/kg bw by
- Amount of vehicle (if gavage): No data available
- Lot/batch no. (if required): No data available
- Purity: No data available
Details on mating procedure:
Study 2:
- M/F ratio per cage: one male and one female
- Length of cohabitation: Female were placed with same male until pregnancy occurs or two weeks elapsed.
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy: The day of detection of sperm positive vaginal smear was considered as day "0" of gestation. - After ... days of unsuccessful pairing replacement of first male by another male with proven fertility.
- Further matings after two unsuccessful attempts: [no / yes (explain)]
- After successful mating each pregnant female was caged (how): Individually
- Any other deviations from standard protocol: No Data Available

Study 3: Premated female rats were used in this study.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Study 2: Specificity, Linearity, Precision (%RSD), Accuracy (% Recovery) and Homogeneity were analyze by using HPLC-UV.
Duration of treatment / exposure:
Study 2: More than 63 days
Study 3: Day 6 to 15 post coitum (10 Days)
Frequency of treatment:
Study 2: Daily
Study 3: Once Daily
Details on study schedule:
Study 2: Dosing of both sexes were dosed 2 weeks prior to mating, after they were acclimatised for nineteen days and females were screened for normal oestrous cycles (in a 2 weeks pre-treatment period). Pups and dams were sacrificed on day 13 and day 14 respectively. Dams were fasted overnight prior to blood collection. The day of birth (viz. when parturition was completed) was defined as day 0 post-partum. Non – pregnant females were sacrificed on day 49. Dosing was continued in both sexes during the mating period. Males were dosed for 48 days. Daily dosing of the parental females kept continued throughout pregnancy and at least up to the day before sacrifice (Day 13 post-partum). Recovery animals were kept 14 days after the first schedule sacrificed of dams without treatment to detect delayed occurrence, or persistence of, or recovery from toxic effects.
Remarks:
Study 2: 0, 750, 1000, 1250 mg/kg bw.
Study 3: 0 and 1000 mg/kg bw
No. of animals per sex per dose:
Study 2:
Total: 124 animals
0 mg/kg bw: 13 male, 13 female
750 mg/kg bw: 13 male, 13 female
1000 mg/kg bw: 13 male, 13 female
1250 mg/kg bw: 13 male, 13 female
Recovery group:
0 mg/kg bw: 5 male, 5 female
1250 mg/kg bw: 5 male, 5 female

Study 3:
Total: 50 animals
0 mg/kg bw (control group): 25 female rats
1000 mg/kg bw (test group): 25 female rats
Control animals:
yes, concurrent vehicle
Details on study design:
Study 2: - Dose selection rationale: No Data Available
- Rationale for animal assignment (if not random): Randomization was done based on recent body weight, before first dosing. The animals were allocated to the different test groups using the ‘Group Allocation’ function in the MS Excel Add-in “Daniel’s XL Toolbar” (http://xltoolbox.sourceforge.net/). Individual body weights were considered within ± 20% of the groups mean. Details of the randomization was documented in the raw data.
- Other: No Data Available
Positive control:
No Data Available
Parental animals: Observations and examinations:
Study 2: CAGE SIDE OBSERVATIONS: Yes
- Time schedule:twice daily (morning and evening)
- Cage side observations checked in table were included. morbidity and mortality, throughout the acclimatization and study period.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: once a day, preferably at the same time each day considering the peak period of anticipated effects after dosing.
BODY WEIGHT: Yes
- Time schedule for examinations:Males and females were weighed during randomization, on the first day of dosing, at least weekly thereafter, and at termination. During pregnancy, females were weighed on days 0, 7, 14 and 20 and within 24 hours of parturition (day 0 or 1 post-partum), day 4 post-partum and before terminal sacrifice.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data
- Time schedule for examinations:

OTHER:Hematology and clinical biochemistry were examined.

Study 3:
CAGE SIDE OBSERVATIONS: No data
- Time schedule: No data
- Cage side observations checked in table were included. No data

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: No data

BODY WEIGHT: No data
- Time schedule for examinations: No data

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): No data
- 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

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study):No data
- Time schedule for examinations: No data

OTHER: Following reproductive parameters were examined:
Corpora lutea
Implantations
Resorptions (complete, early and late)
Oestrous cyclicity (parental animals):
Study 2: Estrous cycle were monitored daily from beginning of the treatment period until evidence of mating.
Sperm parameters (parental animals):
No Data Available
Litter observations:
Study 2: Number and sex of pups, stillbirths, live births, runts (pups that are significantly smaller than corresponding control pups) were examined.
Postmortem examinations (parental animals):
Study 2:Organ Weight, Gross Pathology and Histopathology were examined.

Study 3: Animals were sacrificed on day 20 and necropsied.
GROSS NECROPSY: Yes
HISTOPATHOLOGY / ORGAN WEIGHTS: Yes
Postmortem examinations (offspring):
Study 2: Gross abnormalities were examined.
Statistics:
Study 2:Raw data was analysed using statistical software “Sigma Plot 11.0” (Supplied by Cranes Software International Ltd. Bangalore). The mean and standard deviation was calculated using the software and all data was summarized in tabular form. All continuous data (body weight, feed consumption, Functional Observational Battery parameters, hematology, clinical chemistry, absolute and relative organ weights, maternal and pup parameters etc.) were checked for normality using Shapiro Wilk test. All homogenous data was analysed using ANOVA and data showing significance in their variances was subjected to Dunnett’s t-test. All heterogeneous data was analysed using F test and Student’s t-test, Dunn’s Test, Kruskal-Wallis, ANOVA on ranks
Reproductive indices:
Study 2: Pregnancy Index, copulation, Post-natal Loss, Pups Survival Index were examined.
Offspring viability indices:
Study 2: Live pups were counted and sexed within 24 hours of parturition (day 0 or 1 post-partum) and on day 4 post-partum.
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
Study 2: No apparent treatment related clinical signs were observed in any of the animals throughout the treatment period. However, statistically significant increase was noted in urine pool in group G2 in male on week 2 as compared to the G1 control group. In recovery female, statistically significant decrease was noted in rearing in G4-R on pretreatment, and increase in fecal bolus on week 1 as compared to the control recovery group G1-R. Statistically significant increase in urine pool in male G2 group on week 2 as compared to the control group G1. Detailed clinical examinations like Home cage observation, Handling observation and Open field observation of all animals were observed to be normal during study period in Main study.The above changes observed were inconsistent/ biologically insignificant and not dose dependant, hence considered as incidental and not attributed to the effect of test item administration.

Study 3: There were no signs of toxicity due to administration of the test chemical in dams.
Dermal irritation (if dermal study):
not specified
Mortality:
mortality observed, non-treatment-related
Description (incidence):
Study 2: No mortality or morbidity was observed in any of the groups of animals throughout the study period however, one male was found dead on Day 42 in group G2 at 750 mg/kg body weight and one male was found dead on Day 21 in group G4 at 1250 mg/kg body weight after dosing due to gavaging error.
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
Study 2: No treatment related changes were noted in body weight and body weight gain upto 1250 mg/kg body weight. However, statistically significant decrease was noted in percent change in body weight with respect to day 1 in G4-R group on day 22 in male as compared to the control recovery group G1-R. Body weight and Percent body weight changes in animals of the all other test groups of both the sexes was comparable and did not show any significant difference as compared to the respective control group. These changes observed were inconsistent, hence not considered as effect of the test item administration.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
Study 2: No treatment related changes were noted upto 1250 mg/kg body weight. However, Feed consumption in animals of the all other test groups of both the sexes was comparable and did not show any significant difference as compared to the respective control group. Changes observed in feed consumption were inconsistent, hence not considered as effect of the test item administration
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Study 2: No treatment related changes were noted in hematological parameters upto 1250 mg/kg body weight. However, statistically significant decreased was noted in male in aPTT in Group G4 at 1250 mg/kg body weight. In recovery males, statistically significant increase were noted in RBC and platelets at 1250 mg/kg body weight and statistically significant decrease was noted in PT at 1250 mg/kg body weight. The observed variations in aPTT, PT, RBC, and platelets were considered to be of no toxicological significance, of a minimal in nature and occurred in the absence of clear dose related response
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
Study 2: No treatment related chages were noted in clinical chemistry parameters upto 1250 mg/kg body weight. However, statistically significant decrease were noted in Albumin (Male: G3 at 1000 mg/kg body weight), creatinine (Male: G3 and G4 at 1000 and 1250 mg/kg body weight respectively), phosphorus (Male: G2 at 750 mg/kg body weight). In recovery males, statistically significant increase was noted in sodium at 1250 mg/kg body weight and statistically significant decrease were noted in AST and phosphorus at 1250 mg/kg body weight. The observed variations in clinical chemistry parameters were considered to be of no toxicological significance, of a minimal in nature and occurred in the absence of clear dose related response No treatment related changes were noted in hormonal analysis (T4, TSH, Testosterone and Estrogen) in any of the groups
Urinalysis findings:
not specified
Behaviour (functional findings):
no effects observed
Description (incidence and severity):
Study 2: The sensory reactivity measurements were comparable and no changes were revealed in any of the animals of all treated groups in both the sexes Foot splay and fore limb and hind limb grip strength parameters were comparable and no treatment related changes were revealed in any of the animals of all treated groups. However, statistically significant increase was noted in male in grip strength of fore limb in G3 group as compared to the control group G1. Motor activity measurements were comparable and no changes were revealed in any of the animals from all treated groups of both the sexes as compare to control groupThe above changes observed were inconsistent, hence considered as incidental and not attributed to the effect of test item administration.
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
Study 2: Lesions observed in liver, kidneys, lungs, heart, adrenals, spleen and reproductive organs of high dose treated group rats are well comparable with respective control group rats and exhibited no dose relationship. Hence, occurrence of these lesions could be considered as spontaneous or incidental in nature and not to be attributed because of administration of the test chemical.
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Reproductive function: oestrous cycle:
no effects observed
Description (incidence and severity):
Study 2: In control group G1 and treatment group G2, G3 and G4 all the females showed regular cyclicity i.e. 3-5 days estrous cycle In cohabitation or mating period, all females from control and treated groups showed evidence of copulation i.e. sperm positive vaginal smear. All females showed precoital interval less than 5 days, except 4 females from G2 which showed precoital interval more than 5 days.
Reproductive function: sperm measures:
not specified
Reproductive performance:
no effects observed
Description (incidence and severity):
Study 2: No statistically significant effect on Gestational length, Litter size, No. of live births, Post-implantation loss, Post-natal loss and Pregnancy index were observed.

Study 3: No effect was observed on numbers of corpora lutea, implantation sites, and viable fetuses were found.
Dose descriptor:
NOAEL
Effect level:
1 250 mg/kg bw/day
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
reproductive function (oestrous cycle)
reproductive performance
Remarks on result:
not determinable due to absence of adverse toxic effects
Critical effects observed:
no
Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality / viability:
no mortality observed
Description (incidence and severity):
Study 2: No statistically significant effect on No. of live births were observed in treated rats as compared to control.

Study 3: No mortality of the fetuses were observed at tested doses.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Study 2: No statistically significant effect on pups weight at birth and PND13.

Study 3: No effects on the fetal body weights were observed.
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Sexual maturation:
not specified
Anogenital distance (AGD):
not specified
Nipple retention in male pups:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Study 2: Terminally sacrificed pups of all treated groups did not reveal any lesion of pathological significance in any of the group when compared with control group. Pups died during course of study revealed various lesions among the control and treated groups viz., external examination Cannibalism (Male: G2:2/20, Female: G2:2/24); and internal examination White Patch on edge of Liver (Male: G2: 1/18).

Study 3: No visceral or external malformations were observed at any given doses.
Histopathological findings:
no effects observed
Description (incidence and severity):
Study 3: Skeletal variations that were observed in the dose groups were similar and comparable to that of the control groups.
Other effects:
not specified
Behaviour (functional findings):
not specified
Developmental immunotoxicity:
not specified
Study 3:Foetuses were weighed, inspected macroscopically for external malformations and prepared for inspection for visceral and skeletal malformations (by transverse section and double staining respectively). They did not show any such malformations. Skeletal variations were the same (nature and frequency) in test and control group.
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
1 250 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
viability
mortality
body weight and weight gain
gross pathology
Remarks on result:
not determinable due to absence of adverse toxic effects
Critical effects observed:
no
Reproductive effects observed:
not specified
Treatment related:
not specified
Conclusions:
Based on all the observations and results, it was concluded that the NOAEL was considered to be 1250 mg/kg bw for P and F1 generation Wistar male and female rat treated with test chemical orally by gavage for more than 63 days.
Executive summary:

Reproductive Toxicity Study:

The available data for the reproductive toxicity studies are as follows:

Study 2:

In a Repeated Dose Oral Toxicity Study in combination with Reproduction / Developmental Toxicity study, Wistar male and female rat treated with test chemical in the concentration of 0, 750, 1000 and 1250 mg/kg bw orally by gavage for more than 63 days. No mortality or morbidity was observed in any of the groups of animals throughout the study period however, one male was found dead on Day 42 at 750 mg/kg body weight and one male was found dead on Day 21 at 1250 mg/kg body weight after dosing due to gavaging error. No apparent treatment related clinical signs were observed in any of the animals throughout the treatment period. However, statistically significant increase was noted in urine pool at 750 mg/kg bw in male on week 2 as compared to control group. In recovery female, statistically significant decrease was noted in rearing at 1250 mg/kg bw on pre treatment, and increase in fecal bolus on week 1 as compared to the control recovery. Detailed clinical examinations like Home cage observation, Handling observation and Open field observation of all animals were observed to be normal during study period in Main study. The above changes observed were inconsistent/ biologically insignificant and not dose dependant, hence considered as incidental and not attributed to the effect of test item administration. No treatment related changes were noted in body weight and body weight gain upto 1250 mg/kg body weight. No treatment related changes were noted upto 1250 mg/kg body weight. However, Feed consumption in animals of the all other test groups of both the sexes was comparable and did not show any significant difference as compared to the respective control group. Changes observed in feed consumption were inconsistent, hence not considered as effect of the test item administration. Similarly, no treatment related changes were noted in hematological parameters upto 1250 mg/kg body weight. There were some observed variations in aPTT, PT, RBC, and platelets which were considered to be of no toxicological significance, of a minimal in nature and occurred in the absence of clear dose related response. No treatment related charges were noted in clinical chemistry parameters upto 1250 mg/kg body weight. The observed variations in clinical chemistry parameters were considered to be of no toxicological significance, of a minimal in nature and occurred in the absence of clear dose related response. No treatment related changes were noted in hormonal analysis (T4, TSH, Testosterone and Estrogen) in any of the groups. The sensory reactivity measurements were comparable and no changes were revealed in any of the animals of all treated groups in both the sexes. Foot splay and fore limb and hind limb grip strength parameters were comparable and no treatment related changes were revealed in any of the animals of all treated groups. In addition, At the end of treatment and recovery period, absolute and relative weight of organs of treated group rats of either sex did not differ significantly when compared to respective control group.  In control group G1 and treatment group G2, G3 and G4 all the females showed regular cyclicity i.e. 3-5 days estrous cycle. In cohabitation or mating period, all females from control and treated groups showed evidence of copulation i.e. sperm positive vaginal smear. All females showed precoital interval less than 5 days, except 4 females at 75 mg/kg bw which showed precoital interval more than 5 days. No differences in mating index, pre and post-implantation losses, resorption index, and sexual functions of the male and female animals were observed at any given doses. In fetal parameters,No statistically significant effect on No. of live births were observed in treated rats as compared to control. No statistically significant effect on pups weight at birth and PND13. Terminally sacrificed pups of all treated groups did not reveal any lesion of pathological significance in any of the group when compared with control group. Pups died during course of study revealed various lesions among the control and treated groups viz., external examination Cannibalism (Male: G2:2/20, Female: G2:2/24); and internal examination White Patch on edge of Liver (Male: G2: 1/18).Therefore, NOAEL was considered to be 1250 mg/kg bw for P and F1 generation Wistar male and female rat treated with test chemical orally by gavage for more than 63 days.

Study 3:

The present study was conducted to determine the reproductive toxicity potential of test chemical, when administered orally to rats. Virgin premated female rats were used for the study. Two dose groups (25 animals each) were used, one test- (1000 mg/kg bw) and one control group (0 mg/kg bw). They were dosed from day 6-15 post coitum and kept off-dose from day 16-19. There were no signs of toxicity in dams. Animals were sacrificed on day 20 and necropsied. Reproductive parameters were examined viz.no. of corpora lutea, no. of implantations and resorptions (complete, early and late). Foetuses were weighed, inspected macroscopically for external malformations and prepared for inspection for visceral and skeletal malformations (by transverse section and double staining respectively). They did not show any such malformations. Skeletal variations were the same (nature and frequency) in test- and control group. No effect was observed on numbers of corpora lutea, implantation sites, and viable fetuses were found. Therefore, under the condition of this study, the no-observed-adverse-effect level (NOAEL) for dams and foetuses was considered to be 1000 mg/kg bw.

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 250 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
Data is from a Klimisch 2 source and provides a robust study summary.
Additional information

Reproductive Toxicity Study:

The available data for the reproductive toxicity studies are as follows:

Study 2:

In a Repeated Dose Oral Toxicity Study in combination with Reproduction / Developmental Toxicity study, Wistar male and female rat treated with test chemical in the concentration of 0, 750, 1000 and 1250 mg/kg bw orally by gavage for more than 63 days. No mortality or morbidity was observed in any of the groups of animals throughout the study period however, one male was found dead on Day 42 at 750 mg/kg body weight and one male was found dead on Day 21 at 1250 mg/kg body weight after dosing due to gavaging error. No apparent treatment related clinical signs were observed in any of the animals throughout the treatment period. However, statistically significant increase was noted in urine pool at 750 mg/kg bw in male on week 2 as compared to control group. In recovery female, statistically significant decrease was noted in rearing at 1250 mg/kg bw on pre treatment, and increase in fecal bolus on week 1 as compared to the control recovery. Detailed clinical examinations like Home cage observation, Handling observation and Open field observation of all animals were observed to be normal during study period in Main study. The above changes observed were inconsistent/ biologically insignificant and not dose dependant, hence considered as incidental and not attributed to the effect of test item administration. No treatment related changes were noted in body weight and body weight gain upto 1250 mg/kg body weight. No treatment related changes were noted upto 1250 mg/kg body weight. However, Feed consumption in animals of the all other test groups of both the sexes was comparable and did not show any significant difference as compared to the respective control group. Changes observed in feed consumption were inconsistent, hence not considered as effect of the test item administration. Similarly, no treatment related changes were noted in hematological parameters upto 1250 mg/kg body weight. There were some observed variations in aPTT, PT, RBC, and platelets which were considered to be of no toxicological significance, of a minimal in nature and occurred in the absence of clear dose related response. No treatment related charges were noted in clinical chemistry parameters upto 1250 mg/kg body weight. The observed variations in clinical chemistry parameters were considered to be of no toxicological significance, of a minimal in nature and occurred in the absence of clear dose related response. No treatment related changes were noted in hormonal analysis (T4, TSH, Testosterone and Estrogen) in any of the groups. The sensory reactivity measurements were comparable and no changes were revealed in any of the animals of all treated groups in both the sexes. Foot splay and fore limb and hind limb grip strength parameters were comparable and no treatment related changes were revealed in any of the animals of all treated groups. In addition, At the end of treatment and recovery period, absolute and relative weight of organs of treated group rats of either sex did not differ significantly when compared to respective control group.  In control group G1 and treatment group G2, G3 and G4 all the females showed regular cyclicity i.e. 3-5 days estrous cycle. In cohabitation or mating period, all females from control and treated groups showed evidence of copulation i.e. sperm positive vaginal smear. All females showed precoital interval less than 5 days, except 4 females at 75 mg/kg bw which showed precoital interval more than 5 days. No differences in mating index, pre and post-implantation losses, resorption index, and sexual functions of the male and female animals were observed at any given doses. In fetal parameters,No statistically significant effect on No. of live births were observed in treated rats as compared to control. No statistically significant effect on pups weight at birth and PND13. Terminally sacrificed pups of all treated groups did not reveal any lesion of pathological significance in any of the group when compared with control group. Pups died during course of study revealed various lesions among the control and treated groups viz., external examination Cannibalism (Male: G2:2/20, Female: G2:2/24); and internal examination White Patch on edge of Liver (Male: G2: 1/18).Therefore, NOAEL was considered to be 1250 mg/kg bw for P and F1 generation Wistar male and female rat treated with test chemical orally by gavage for more than 63 days.

Study 3:

The present study was conducted to determine the reproductive toxicity potential of test chemical, when administered orally to rats. Virgin premated female rats were used for the study. Two dose groups (25 animals each) were used, one test- (1000 mg/kg bw) and one control group (0 mg/kg bw). They were dosed from day 6-15 post coitum and kept off-dose from day 16-19. There were no signs of toxicity in dams. Animals were sacrificed on day 20 and necropsied. Reproductive parameters were examined viz.no. of corpora lutea, no. of implantations and resorptions (complete, early and late). Foetuses were weighed, inspected macroscopically for external malformations and prepared for inspection for visceral and skeletal malformations (by transverse section and double staining respectively). They did not show any such malformations. Skeletal variations were the same (nature and frequency) in test- and control group. No effect was observed on numbers of corpora lutea, implantation sites, and viable fetuses were found. Therefore, under the condition of this study, the no-observed-adverse-effect level (NOAEL) for dams and foetuses was considered to be 1000 mg/kg bw.

Justification for classification or non-classification

Based on all the available data, it was concluded that the test chemical did not have any effects and reproductive health of the test animals and thus is not likely to classify as a 'Reproductive Toxicant' as per CLP regulation.

Additional information