Registration Dossier

Toxicological information

Endpoint summary

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Administrative data

Key value for chemical safety assessment

Toxic effect type:
dose-dependent

Effects on fertility

Description of key information

Reproductive toxicity study :

NOAEL for the test chemical for systemic and reproductive toxicity was established at 1250 mg/kg bw/day in male and female rats following oral gavage exposure before mating, during mating and thereafter for a total of 48 to 63 days.

Link to relevant study records
Reference
Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Data is from study report
Qualifier:
according to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
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 Identification: During 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. Bedding material of batch No. 4-17 and 817 (Krishana Corncob Industries, Aurangabad)) was used in this study and a copy of report of microbial and chemical contaminants analysed periodically by manufacturer of bedding material are incorporated in the raw data.
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
Type of inhalation exposure (if applicable):
not specified
Vehicle:
corn oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:The test item was weighed and dissolved in a vehicle (corn oil) to achieve desired concentration of test item. Dose formulation was freshly prepared daily. At the time of dosing, dose formulation was kept on the magnetic stirrer to maintain the homogeneity of test item. The details of dose formulation preparation is maintained in raw data.

DIET PREPARATION - Rate of preparation of diet (frequency): - Mixing appropriate amounts with (Type of food):
- Storage temperature of food:
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:
Details on mating procedure:
- 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:
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Specificity, Linearity, Precision (%RSD), Accuracy (% Recovery) and Homogeneity were analyze by using HPLC-UV.
Duration of treatment / exposure:
48 days (male rats, main study)
Approx. 63 days (female rats, main study)
66 days (male and female rats of the reocvery groups)
Frequency of treatment:
Daily
Details on study schedule:
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.
Dose / conc.:
0 mg/kg bw/day
Remarks:
G1 (main study), G1-R (recovery group)
Dose / conc.:
750 mg/kg bw/day
Remarks:
G2
Dose / conc.:
1 000 mg/kg bw/day
Remarks:
G3
Dose / conc.:
1 250 mg/kg bw/day
Remarks:
G4 (main study), G4-R (recovery group)
No. of animals per sex per dose:
13 per sex per dose level (main study)
5 per sex per dose level (reocery groups)
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: - 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:
Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes - Time schedule:twice daily (morning and evening) - Cage side observations checked in table [No.?] 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.
Oestrous cyclicity (parental animals):
Estrous cycle were monitored daily from beginning of the treatment period until evidence of mating.
Sperm parameters (parental animals):
not specified
Litter observations:
Number and sex of pups, stillbirths, live births, runts (pups that are significantly smaller than corresponding control pups) were examined.
Postmortem examinations (parental animals):
Organ weight, gross Pathology and histopathology
Postmortem examinations (offspring):
Gross abnormalities were examined.
Statistics:
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:
Pregnancy index, copulation, post-natal loss, and survival index
Offspring viability indices:
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):
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 test chemical.
Dermal irritation (if dermal study):
not examined
Mortality:
mortality observed, non-treatment-related
Description (incidence):
No mortality was observed other than two deaths due to gavage error (one male at 750 and one male and 1000 mg/kg).
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
Body weight changes were restricted to a significant decrease in % body weight change in the recovery group of male rats treated at 1250 mg/kg from day 1 – 22 as compared to the control group. This effect was considered incidental and therefore not attributed to the test chemical.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
Food intake was unaffected by treatment.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Changes in haematology included a significant decrease in activated partial thromboplastin time in male rats treated at 1250 mg/kg compared to controls; a significant increase in red blood cell levels in the recovery group of male rats treated at 1250 mg/kg compared to controls; a significant increase in platelet levels in the recovery group of male rats treated at 1250 mg/kg compared to controls; and a significant increase in prothrombin time in the recovery group of male rats treated at 1250 mg/kg compared to controls. The changes in haematology were not consistent between groups and lacked histological correlates. Therefore, the changes in haematology and clinical chemistry were not considered to be of toxicological importance.
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
Changes in clinical chemistry included decreased creatinine levels in male rats treated at ≥1000 mg/kg compared to controls; increased sodium levels in the recovery group of male rats treated at 1250 mg/kg compared to controls; decreased phosphorus levels in the recovery group of male rats treated at 1250 mg/kg compared to controls; decreased aspartate transaminase levels in the recovery groups of male and female rats treated at 1250 mg/kg compared to controls. The changes in clinical chemistry were not consistent between groups and lacked histological correlates. Therefore, the changes in clinical chemistry were not considered to be of toxicological importance.
Urinalysis findings:
not examined
Behaviour (functional findings):
effects observed, non-treatment-related
Description (incidence and severity):
Changes in motor activity and behaviour were restricted to a significant increase in grip strength in male rats treated at 1000 mg/kg bw/day as compared to control. This effect was considered incidental and therefore not attributed to the test chemical.
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
Liver: multifocal minimal to mild Neutrophil/lymphocyte infiltration (Male: G1:2/5, G4:1/5; Female: G1/5), focal to multifocal mild degeneration (Male: G1:1/5, Female: G4/5), Kidneys: focal to multifocal mild lymphocyte infiltration (Male: G1:2/5, G4:1/5; Female: G1/5, G4:1/5); Lungs: multifocal minimal to mild lymphocyte infiltration (Male: G1:1/5, G4:1/5; Female: G1/5, G4:1/5), Heart: focal to multifocal minimal lymphocyte infiltration (Male: G1:1/5; Female: G4:1/5), Spleen: multifocal mild Extramedullary haematopoiesis (Male: G1:1/5), Adrenals: multifocal mild cytoplasmic vacuolation (male: G1:1/5), accessory adrenocortical tissue (Unilateral: Male: G1:2/5, G4:2/5; Female: G1:1/5), Testes: Male: focal mild seminiferous tubule degeneration (Male: G1: 2/13, G2:2/13, G4: 2/13, G1-R: 1/5); focal minimal to mild retention of mature sperm (Male: G1: 1/13, G2:1/13, G3: 1/13, G4: 1/13); focal minimal to mild sloughing of round spermatid/pachytene spermatocyte (Male: G1: 1/13, G2:1/13). 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. Further these observed lesions are common in occurrence in rodents during toxicological studies (Boorman et al., 1990, Greaves, 2007, Greaves and Faccini, 1992; Haschek et al., 2002). 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 Item
Histopathological findings: neoplastic:
no effects observed
Other effects:
no effects observed
Description (incidence and severity):
Hormonal data showed no significant effects on the concentrations of T4 or TSH (male and females), testosterone (males) or oestradiol (females).
Reproductive function: oestrous cycle:
no effects observed
Description (incidence and severity):
Oestrous cyclicity was unaffected by treatment and all female rats showed evidence of copulation after the cohabitation/mating period. All females showed precoital intervals of less than 5 days except for four female rats at 750 mg/kg who showed intervals of more than 5 days.
Reproductive function: sperm measures:
not examined
Reproductive performance:
no effects observed
Description (incidence and severity):
Pregnancy rates were 77, 62, 77 and 77% at 0, 750, 1000 and 1250 mg/kg, respectively. No significant effects were observed on gestation length or litter size. Likewise, no significant effects were observed on the number of live births.
Key result
Dose descriptor:
NOAEL
Effect level:
>= 1 250 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
haematology
clinical biochemistry
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
histopathology: neoplastic
reproductive function (oestrous cycle)
reproductive performance
Remarks on result:
other: No effect observed
Critical effects observed:
no
System:
other: not specified
Organ:
not specified
Clinical signs:
not specified
Dermal irritation (if dermal study):
not examined
Mortality / viability:
no mortality observed
Description (incidence and severity):
No statistically significant effect on number of live births were observed in treated rats as compared to controls. However four pups were eventually cannibalized in the 750 mg/kg treatment group.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No statistically significant effect on pups weight at birth or PND13
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Sexual maturation:
not examined
Anogenital distance (AGD):
not examined
Nipple retention in male pups:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Four pups were cannibalized at the 750 mg/kg treatment group. All other pups at 0, 750, 1000 and 1250 mg/kg were normal externally. The internal examination of the pups revealed no abnormalities attributed to the test chemical
Histopathological findings:
no effects observed
Description (incidence and severity):
Microscopic examination of the pups’ thyroid and parathyroid glands at 0 and 1250 mg/kg revealed no abnormal findings.
Other effects:
no effects observed
Description (incidence and severity):
No significant effect on sex ratio was observed.
Behaviour (functional findings):
not specified
Developmental immunotoxicity:
not specified
Key result
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
>= 1 250 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
viability
mortality
body weight and weight gain
gross pathology
histopathology: non-neoplastic
Remarks on result:
other: No effect observed
Critical effects observed:
no
System:
other: not specified
Organ:
not specified
Reproductive effects observed:
no

Mortality and Morbidity

Sex: Male

Group

Treatment

Dose (mg/kg b.wt.)

No. of Animals/

Group

Observation During Study Period

G1

Control

0

13

NMM

G2

Low

750

13

01/13 (FD on Day 42)

G3

Mid

1000

13

01/13 (FD on Day 21)

G4

High

1250

13

NMM

G1-R

Control- Recovery

0

5

NMM

G4-R

High- Recovery

1250

5

NMM

 Sex: Female

Group

Treatment

Dose (mg/kg b.wt.)

No. of Animals/

Group

Observation During Study Period

G1

Control

0

13

NMM

G2

Low

750

13

NMM

G3

Mid

1000

13

NMM

G4

High

1250

13

NMM

G1-R

Control -Recovery

0

5

NMM

G4-R

High- Recovery

1250

5

NMM

Keys:NMM = No mortality and morbidity observed, No.= Number, FD= Found dead after dosing due to gavaging error.

MeanHormonal Analysis Data

 Sex: Male

Group

G1

G2

G3

G4

Dose(mg/kg bwt)

0

750

1000

1250

Mean

SD

N

Mean

SD

N

Mean

SD

N

Mean

SD

N

T4 (ug/dL)

3.33

0.55

13

4.24

2.66

13

4.28

2.67

13

3.62

0.66

13

TSH (uIU/mL)

2.16

1.47

13

4.55

4.01

13

2.69

3.22

13

2.38

1.65

13

Testosterone (ng/dL)

185.98

147.96

13

188.00

192.69

13

148.34

187.15

13

169.54

140.02

13

 Sex: Female

Group

G1

G2

G3

G4

Dose(mg/kg bwt)

0

750

1000

1250

Mean

SD

N

Mean

SD

N

Mean

SD

N

Mean

SD

N

T4 (ug/dL)

3.66

0.56

10

3.81

1.02

8

3.35

0.44

10

3.75

0.58

10

TSH (uIU/mL)

3.45

1.05

10

3.49

2.52

8

2.21

0.93

10

3.26

1.03

10

E2 (pg/mL)

 

24.26

17.29

10

45.51

31.42

8

23.88

6.77

10

31.09

16.46

10

 Sex: Female (Non-Pregnenat)

Group

G1

G2

G3

G4

Dose(mg/kg bwt)

0

750

1000

1250

Mean

SD

N

Mean

SD

N

Mean

SD

N

Mean

SD

N

T4 (ug/dL)

3.47

0.15

3

3.24

 

0.43

5

3.57

 

0.40

3

3.07

0.35

3

TSH (uIU/mL)

1.63

0.21

3

2.22

1.28

5

1.81

0.88

3

0.93

0.27

3

E2 (pg/mL)

 

38.06

5.98

3

34.73

26.87

5

19.28

0.83

2

34.69

14.33

3

Sex: Male

Group(n)

G1R

G4R

Dose(mg/kg bwt)

0

1250

Mean

SD

N

Mean

SD

N

T4 (ug/dL)

2.86

0.79

5

3.40

0.85

5

TSH (uIU/mL)

3.04

2.08

5

1.21

1.16

5

Testosterone (ng/dL)

20.95

10.16

5

90.59

84.78

5

 Sex: Female

 

Group(n)

G1R

G4R

Dose(mg/kg bwt)

0

1250

Mean

SD

N

Mean

SD

N

T4 (ug/dL)

2.62

 

0.31

5

2.58

0.54

5

TSH (uIU/mL)

0.86

0.42

5

2.43

0.83

5

Testosterone (ng/dL)

20.69

7.96

4

21.31

7.49

5

Day: 04 

Group

G1

G2

G3

G4

Dose(mg/kg bwt)

0

750

1000

1250

 

Mean

SD

N

Mean

SD

N

Mean

SD

N

Mean

SD

N

T4 (ug/dL)

2.03

0.31

6

2.53

0.82

4

1.96

0.32

9

2.21

0.31

8

TSH (uIU/mL)

1.42

0.56

8

1.79

0.95

4

1.26

0.55

10

1.88

0.59

10

Day: 13

Group(n)

G1

G2

G3

G4

Dose(mg/kg bwt)

0

750

1000

1250

 

Mean

SD

N

Mean

SD

N

Mean

SD

N

Mean

SD

N

T4 (ug/dL)

5.49

1.16

9

5.40

0.98

5

5.34

1.03

10

5.91

1.66

9

TSH (uIU/mL)

1.87

0.93

9

2.52

1.16

5

1.98

0.95

10

2.49

2.68

10

Summary of Days of Conception and Pregnancy Index (%)

Key:N= number of dams in a group, No. = Number


Conclusions:
NOAEL for the test chemical for systemic and reproductive toxicity was established at 1250 mg/kg bw/day in male and female rats following oral gavage exposure before mating, during mating and thereafter for a total of 48 to 63 days.
Executive summary:

The chemical was given to 13 rats per sex per dose level at 0, 750, 1000 and 1250 mg/kg bw/day by oral gave. Male rats were treated two weeks before mating and thereafter for a total of 48 dosing days. Female rats were treated two weeks before mating, during mating, during gestation and during lactation, for a total of approx. 63 dosing days. No morbidity was observed during the study period. No mortality was observed other than two deaths due to gavage error (one male at 750 and one male and 1000 mg/kg). Clinical findings were sporadic and of no biological significance. Body weight changes were restricted to a significant decrease in % body weight change in the recovery group of male rats treated at 1250 mg/kg from day 1 – 22 as compared to the control group. As such, no significant body weight effects were observed for female rats during the study period. Food intake was unaffected by treatment. Hormonal data showed no significant effects on the concentrations of T4 or TSH (male and females), testosterone (males) or oestradiol (females). Oestrous cyclicity was unaffected by treatment and all female rats showed evidence of copulation after the cohabitation/mating period. All females showed precoital intervals of less than 5 days except for four female rats at 750 mg/kg who showed intervals of more than 5 days. Pregnancy rates were 77, 62, 77 and 77% at 0, 750, 1000 and 1250 mg/kg, respectively. No significant effects were observed on gestation length or litter size. Likewise, no significant effects were observed on the number of live births, pup survival, pup weight or sex ratio. Four pups were cannibalized at the 750 mg/kg treatment group. All other pups at 0, 750, 1000 and 1250 mg/kg were normal externally. The internal examination of the pups revealed no abnormalities attributed to the test chemical. Microscopic examination of the pups’ thyroid and parathyroid glands at 0 and 1250 mg/kg revealed no abnormal findings. In the adult rats, no significant effects were observed on the absolute or relative weights of the testes or epididymides. Grossly, one rat treated at 1250 mg/kg showed a mild reduction in the size of the testes. Organ weight data was not collected for the prostate and seminal vesicles with coagulating glands. Gross and microscopic examination of ovaries, uterus, cervix with vagina, testes, epididymides, prostate, seminal vesicle with coagulating glands at 0 and 1250 mg/kg revealed no significant effects attributed to the test chemical. In the testes, mild focal seminiferous tubular degeneration was observed in 2/13, 2/13, 0/13 and 2/13 rats at 0, 750, 1000 and 1250 mg/kg, respectively. Retention of mature sperm (minimal or mild) was observed in 1/13 rats in all treatment groups. Likewise, sloughing of round spermatids/pachytene spermatocytes was observed in 1/13 rats in all treatment groups. NOAEL for the test chemical for systemic and reproductive toxicity was established at 1250 mg/kg bw/day in male and female rats following oral gavage exposure before mating, during mating and thereafter for a total of 48 to 63 days.

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 Klimisch 1 and from study report
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

Reproductive toxicity study :

In a study, test chemical has been investigated for reproductive toxicity. The study is summarized below 

The chemical was given to 13 rats per sex per dose level at 0, 750, 1000 and 1250 mg/kg bw/day by oral gave. Male rats were treated two weeks before mating and thereafter for a total of 48 dosing days. Female rats were treated two weeks before mating, during mating, during gestation and during lactation, for a total of approx. 63 dosing days. No morbidity was observed during the study period. No mortality was observed other than two deaths due to gavage error (one male at 750 and one male and 1000 mg/kg). Clinical findings were sporadic and of no biological significance. Body weight changes were restricted to a significant decrease in % body weight change in the recovery group of male rats treated at 1250 mg/kg from day 1 – 22 as compared to the control group. As such, no significant body weight effects were observed for female rats during the study period. Food intake was unaffected by treatment. Hormonal data showed no significant effects on the concentrations of T4 or TSH (male and females), testosterone (males) or oestradiol (females). Oestrous cyclicity was unaffected by treatment and all female rats showed evidence of copulation after the cohabitation/mating period. All females showed precoital intervals of less than 5 days except for four female rats at 750 mg/kg who showed intervals of more than 5 days. Pregnancy rates were 77, 62, 77 and 77% at 0, 750, 1000 and 1250 mg/kg, respectively. No significant effects were observed on gestation length or litter size. Likewise, no significant effects were observed on the number of live births, pup survival, pup weight or sex ratio. Four pups were cannibalized at the 750 mg/kg treatment group. All other pups at 0, 750, 1000 and 1250 mg/kg were normal externally. The internal examination of the pups revealed no abnormalities attributed to the test chemical. Microscopic examination of the pups’ thyroid and parathyroid glands at 0 and 1250 mg/kg revealed no abnormal findings. In the adult rats, no significant effects were observed on the absolute or relative weights of the testes or epididymides. Grossly, one rat treated at 1250 mg/kg showed a mild reduction in the size of the testes. Organ weight data was not collected for the prostate and seminal vesicles with coagulating glands. Gross and microscopic examination of ovaries, uterus, cervix with vagina, testes, epididymides, prostate, seminal vesicle with coagulating glands at 0 and 1250 mg/kg revealed no significant effects attributed to the test chemical. In the testes, mild focal seminiferous tubular degeneration was observed in 2/13, 2/13, 0/13 and 2/13 rats at 0, 750, 1000 and 1250 mg/kg, respectively. Retention of mature sperm (minimal or mild) was observed in 1/13 rats inall treatment groups. Likewise, sloughing of round spermatids/pachytene spermatocytes was observed in 1/13 rats in all treatment groups.NOAEL for the test chemical for systemic and reproductive toxicity was established at 1250 mg/kg bw/day in male and female rats following oral gavage exposure before mating, during mating and thereafter for a total of 48 to 63 days.

Effects on developmental toxicity

Description of key information

Developmental toxicity study:

NOAEL for the test chemical for systemic and developmental toxicity was established at 1250 mg/kg bw/day in male and female rats following oral gavage exposure before mating, during mating and thereafter for a total of 48 to 63 days.

Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Data is from study report
Qualifier:
according to
Guideline:
other: OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Wistar
Details on test animals and environmental conditions:
Source: Animals were procured from a CPCSEA approved Vendor [National Institute of Biosciences, PuneHealth Status: Healthy young adult animals were used for the study. Females were nulliparous and nonpregnant.
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.Bedding material of batch No. 4-17 and 817 (Krishana Corncob Industries, Aurangabad)) was used in this study and a copy of report of microbial and chemical contaminants analysed periodically by manufacturer of bedding material are incorporated in the raw data.
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. The copy of composition, WaterAqua guard filtered drinking water in bottles was offered ad libitum. Samples of the drinking water was subjected periodically to bacteriological tests and to chemical contaminant analysis. The latest test results are included in the raw data.
Route of administration:
oral: gavage
Type of inhalation exposure (if applicable):
not specified
Vehicle:
corn oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:The test item was weighed and dissolved in a vehicle (corn oil) to achieve desired concentration of test item. Dose formulation was freshly prepared daily. At the time of dosing, dose formulation was kept on the magnetic stirrer to maintain the homogeneity of test item. The details of dose formulation preparation is maintained in raw data.
DIET PREPARATION- Rate of preparation of diet (frequency):
- Mixing appropriate amounts with (Type of food):
- Storage temperature of food:
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:
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Specificity, Linearity, Precision (%RSD), Accuracy (% Recovery) and Homogeneity were analyze by using HPLC-UV.
Details on mating procedure:
- 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:
Duration of treatment / exposure:
Males: 48 days (main study)
Females: 63 days (main study)
Male and females of recovery groups: 66 days
Frequency of treatment:
Daily
Duration of test:
48 days (male rats, main study)
Approx 63 days (female rats, main study)
66 days + 14 days of recovery (recovery groups).
Dose / conc.:
0 mg/kg bw/day (actual dose received)
Remarks:
G1 (main study), G1-R (recovery group)
Dose / conc.:
750 mg/kg bw/day (actual dose received)
Remarks:
G2
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
Remarks:
G3
Dose / conc.:
1 250 mg/kg bw/day (actual dose received)
Remarks:
G4 (main study), G4-R (recovery group)
No. of animals per sex per dose:
13 rats per sex per dose level (main study)
5 rats per sex per dose level (recovery groups)
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale:- 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 Allocati on’ 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:
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes- Time schedule:twice daily (morning and evening)- Cage side observations checked in table [No.?] 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, Organ Weight, Gross Pathology and Histopathology were examined.
Ovaries and uterine content:
Oestrous cyclicity and gestation length
Fetal examinations:
Number and sex of pups, stillbirths, live births, runts (pups that are significantly smaller than corresponding control pups), and gross abnormalities.
Statistics:
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
Indices:
Pregnancy index, copulation, post-natal loss, and pups survival Index.
Historical control data:
Not specified
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
No apparent treatment related clinical signs were observed in any of the animals throughout the treatment 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 obser vation of all animals were observed to be normal during study period in the 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
Dermal irritation (if dermal study):
not examined
Mortality:
mortality observed, non-treatment-related
Description (incidence):
No mortality was observed other than two deaths due to gavage error (one male at 750 and one male and 1000 mg/kg).
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
Body weight changes were restricted to a significant decrease in % body weight change in the recovery group of male rats treated at 1250 mg/kg from day 1 – 22 as compared to the control group. This effect was considered incidental and therefore not attributed to the test chemical.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
Food intake was unaffected by treatment.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Changes in haematology included a significant decrease in activated partial thromboplastin time in male rats treated at 1250 mg/kg compared to controls; a significant increase in red blood cell levels in the recovery group of male rats treated at 1250 mg/kg compared to controls; a significant increase in platelet levels in the recovery group of male rats treated at 1250 mg/kg compared to controls; and a significant increase in prothrombin time in the recovery group of male rats treated at 1250 mg/kg compared to controls. The changes in haematology were not consistent between groups and lacked histological correlates. Therefore, the changes in haematology were not considered to be of toxicological importance.
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
Changes in clinical chemistry included decreased creatinine levels in male rats treated at ≥1000 mg/kg compared to controls; increased sodium levels in the recovery group of male rats treated at 1250 mg/kg compared to controls; decreased phosphorus levels in the recovery group of male rats treated at 1250 mg/kg compared to controls; decreased aspartate transaminase levels in the recovery groups of male and female rats treated at 1250 mg/kg compared to controls. The changes in clinical chemistry were not consistent between groups and lacked histological correlates. Therefore, the changes in clinical chemistry were not considered to be of toxicological importance.
Urinalysis findings:
not examined
Behaviour (functional findings):
effects observed, non-treatment-related
Description (incidence and severity):
Changes in motor activity and behaviour were restricted to a significant increase in grip strength in male rats treated at 1000 mg/kg bw/day as compared to control. This effect was considered incidental and therefore not attributed to the test chemical.
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
There were no significant effects on either the absolute or the relative weight of the brain, adrenals, heart, liver, kidneys, spleen, thymus, thyroid with parathyroid, testes, or epididymides.
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
All adult animals were normal externally. Visceral internal fidnings included a case of mild splenic enlargement at 1000 mg/kg and one case of mild testicular shrinkage at 1250 mg/kg.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
Liver: multifocal minimal to mild Neutrophil/lymphocyte infiltration (Male: G1:2/5, G4:1/5; Female: G1/5), focal to multifocal mild degeneration (Male: G1:1/5, Female: G4/5), Kidneys: focal to multifocal mild lymphocyte infiltration (Male: G1:2/5, G4:1/5; Female: G1/5, G4:1/5); Lungs: multifocal minimal to mild lymphocyte infiltration (Male: G1:1/5, G4:1/5; Female: G1/5, G4:1/5), Heart: focal to multifocal minimal lymphocyte infiltration (Male: G1:1/5; Female: G4:1/5), Spleen: multifocal mild Extramedullary haematopoiesis (Male: G1:1/5), Adrenals: multifocal mild cytoplasmic vacuolation (male: G1:1/5), accessory adrenocortical tissue (Unilateral: Male: G1:2/5, G4:2/5; Female: G1:1/5), Testes: Male: focal mild seminiferous tubule degeneration (Male: G1: 2/13, G2:2/13, G4: 2/13, G1-R: 1/5); focal minimal to mild retention of mature sperm (Male: G1: 1/13, G2:1/13, G3: 1/13, G4: 1/13); focal minimal to mild sloughing of round spermatid/pachytene spermatocyte (Male: G1: 1/13, G2:1/13). 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 relation ship. Further these observed lesions are common in occurrence in rodents during toxicological studies (Boorman et al., 1990, Greaves, 2007, Greaves and Faccini, 1992; Haschek et al., 2002). 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 Item
Histopathological findings: neoplastic:
no effects observed
Other effects:
no effects observed
Description (incidence and severity):
Hormonal data showed no significant effects on the concentrations of T4 or TSH (male and females), testosterone (males) or oestradiol (females).
Number of abortions:
not specified
Pre- and post-implantation loss:
no effects observed
Description (incidence and severity):
No significant effects on post-implnatation loss was observed.
Total litter losses by resorption:
not specified
Early or late resorptions:
not specified
Dead fetuses:
no effects observed
Changes in pregnancy duration:
no effects observed
Changes in number of pregnant:
no effects observed
Other effects:
not specified
Details on maternal toxic effects:
Oestrous cyclicity was unaffected by treatment and all female rats showed evidence of copulation after the cohabitation/mating period. All females showed precoital intervals of less than 5 days except for four female rats at 750 mg/kg who showed intervals of more than 5 days. Pregnancy rates were 77, 62, 77 and 77% at 0, 750, 1000 and 1250 mg/kg, respectively. No significant effects were observed on gestation length or litter size. Likewise, no significant effects were observed on the number of live births. Post-implantation loss was also unaffected by treatment.
Key result
Dose descriptor:
NOAEL
Effect level:
>= 1 250 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
haematology
clinical biochemistry
behaviour (functional findings)
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
histopathology: neoplastic
pre and post implantation loss
effects on pregnancy duration
dead fetuses
changes in pregnancy duration
changes in number of pregnant
necropsy findings
Remarks on result:
other: No effect observed
Abnormalities:
no effects observed
Localisation:
not specified
Description (incidence and severity):
not specified
Fetal body weight changes:
no effects observed
Description (incidence and severity):
No statistically significant effect on pups weight at birth or on PND13
Reduction in number of live offspring:
no effects observed
Description (incidence and severity):
No statistically significant effect at birth or at PND 13
Changes in sex ratio:
no effects observed
Description (incidence and severity):
No significant effects observed.
Changes in litter size and weights:
no effects observed
Description (incidence and severity):
No statistically significant effect on litter size at birth or PND13
Changes in postnatal survival:
no effects observed
Description (incidence and severity):
No significant effects on post-natal loss were observed in treated rats.
External malformations:
effects observed, non-treatment-related
Description (incidence and severity):
Four pups were cannibalized at the 750 mg/kg treatment group. All other pups at 0, 750, 1000 and 1250 mg/kg were normal externally. The internal examination of the pups revealed no abnormalities attributed to the test chemical.
Skeletal malformations:
not examined
Visceral malformations:
not examined
Other effects:
not specified
Key result
Dose descriptor:
NOAEL
Effect level:
>= 1 250 mg/kg bw/day (actual dose received)
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
changes in postnatal survival
external malformations
Remarks on result:
other: No effect observed
Abnormalities:
no effects observed
Localisation:
other: not specified
Description (incidence and severity):
not specified
Developmental effects observed:
no
Treatment related:
not specified

Mortality and Morbidity

Sex: Male

Group

Treatment

Dose (mg/kg b.wt.)

No. of Animals/

Group

Observation During Study Period

G1

Control

0

13

NMM

G2

Low

750

13

01/13 (FD on Day 42)

G3

Mid

1000

13

01/13 (FD on Day 21)

G4

High

1250

13

NMM

G1-R

Control- Recovery

0

5

NMM

G4-R

High- Recovery

1250

5

NMM

 

Sex: Female

Group

Treatment

Dose (mg/kg b.wt.)

No. of Animals/

Group

Observation During Study Period

G1

Control

0

13

NMM

G2

Low

750

13

NMM

G3

Mid

1000

13

NMM

G4

High

1250

13

NMM

G1-R

Control -Recovery

0

5

NMM

G4-R

High- Recovery

1250

5

NMM

Keys:NMM = No mortality and morbidity observed, No.= Number, FD= Found dead after dosing due to gavaging error.

Summary of Days of Conception and Pregnancy Index (%)

 

Group(N)

G1(13)

G2(13)

G3(13)

G4(13)

Dose (mg/kg b.wt.)

0

750

1000

1250

No. of females showed evidence of copulation

13

13

13

13

No. of females concieved between Days 1-5 of cohabitation

13

9

13

13

No. of females concieved after Day 5 of cohabitation

0

4

0

0

Females achieved pregnancy

10

8

10

10

Pregnancy Index (%)

76.92

61.54

76.92

76.92

Key:N= number of dams in a group, No. = Number


Mean Gestational Length and Litter size

 

Group(N)

G1(10)

G2(8)

G3(10)

G4(10)

Dose(mg/kg bwt)

0

750

1000

1250

Parameter

Mean

SD

Mean

SD

Mean

SD

Mean

SD

Gestation Length

22.30

0.48

23.50

1.60

22.10

0.32

22.10

0.32

Litter size

(Total No. of litter size)

10.90

2.18

6.38

4.50

9.90

3.03

9.70

3.62

Keys:SD= Standard Deviation, N= number of dams in a group

Note: Three female was non-pregnant in Group G1, G3, G4 and five females were non-pregnant in G2 group.

Mean Pups Body Weight, Sex Ratio and Gross Observation

 

Group

G1

G2

G3

G4

Dose(mg/kg bwt)

0

750

1000

1250

Mean Pups Weight

Mean

SD

N

Mean

SD

N

Mean

SD

N

Mean

SD

N

Day 0

6.43

0.92

94

6.54

0.46

44

6.00

0.84

96

6.20

0.59

97

Day 4

9.35

1.21

90

8.99

1.07

38

9.36

1.13

91

9.11

1.28

87

Day 13

22.83

3.34

72

20.42

2.37

29

23.46

2.64

68

23.14

3.03

64

Group(n)

G1(10)

G2(8)

G3(10)

G4(10)

Pups Body Weight gain

(%)

Mean

SD

N

Mean

SD

N

Mean

SD

N

Mean

SD

N

Day 0-Day 4

43.96

13.72

90

33.65

9.39

38

55.96

18.26

91

38.77

9.18

87

Day 0-Day 13

140.09

12.44

72

129.99

11.44

29

153.02

12.23

68

134.85

22.05

64

Group

(Number of Litter size)

G1(107)

G2(51)

G3(96)

G4(97)

 Sex Ratio at birth

(Male/Female)

48/46

20/24

49/47

48/49

 Sex Ratio at Day 4

(Male/Female)

40/38

17/21

48/43

44/43

Gross Observations

NAD

NAD

NAD

NAD

Keys:SD= Standard Deviation, N= number of dams with live pups on Day 0, n= number of dams with alive pups on Day 4, NAD = No Abnormality Detected


Conclusions:
NOAEL for systemic and developmental toxicity was established at 1250 mg/kg bw/day in male and female rats. 
Executive summary:

The chemical was given to 13 rats per sex per dose level at 0, 750, 1000 and 1250 mg/kg bw/day by oral gave. Male rats were treated two weeks before mating and thereafter for a total of 48 dosing days. Female rats were treated two weeks before mating, during mating, during gestation and during lactation, for a total of approx. 63 dosing days. No morbidity was observed during the study period. No mortality was observed other than two deaths due to gavage error (one male at 750 and one male and 1000 mg/kg). Clinical findings were sporadic and of no biological significance. Body weight changes were restricted to a significant decrease in % body weight change in the recovery group of male rats treated at 1250 mg/kg from day 1 – 22 as compared to the control group. As such, no significant body weight effects were observed for female rats during the study period. Food intake was unaffected by treatment. Hormonal data showed no significant effects on the concentrations of T4 or TSH (male and females), testosterone (males) or oestradiol (females). Oestrous cyclicity was unaffected by treatment and all female rats showed evidence of copulation after the cohabitation/mating period. All females showed precoital intervals of less than 5 days except for four female rats at 750 mg/kg who showed intervals of more than 5 days. Pregnancy rates were 77, 62, 77 and 77% at 0, 750, 1000 and 1250 mg/kg, respectively. No significant effects were observed on gestation length or litter size. Likewise, no significant effects were observed on the number of live births, pup survival, pup weight or sex ratio. Four pups were cannibalized at the 750 mg/kg treatment group. All other pups at 0, 750, 1000 and 1250 mg/kg were normal externally. The internal examination of the pups revealed no abnormalities attributed to the test chemical. Microscopic examination of the pups’ thyroid and parathyroid glands at 0 and 1250 mg/kg revealed no abnormal findings. In the adult rats, no significant effects were observed on the absolute or relative weights of the testes or epididymides. Grossly, one rat treated at 1250 mg/kg showed a mild reduction in the size of the testes. Organ weight data was not collected for the prostate and seminal vesicles with coagulating glands. Gross and microscopic examination of ovaries, uterus, cervix with vagina, testes, epididymides, prostate, seminal vesicle with coagulating glands at 0 and 1250 mg/kg revealed no significant effects attributed to the test chemical. In the testes, mild focal seminiferous tubular degeneration was observed in 2/13, 2/13, 0/13 and 2/13 rats at 0, 750, 1000 and 1250 mg/kg, respectively. Retention of mature sperm (minimal or mild) was observed in 1/13 rats in all treatment groups. Likewise, sloughing of round spermatids/pachytene spermatocytes was observed in 1/13 rats in all treatment groups. NOAEL for systemic and developmental toxicity was established at 1250 mg/kg bw/day in male and female rats. 

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 250 mg/kg bw/day
Study duration:
chronic
Species:
rat
Quality of whole database:
Data is Klimisch 2 and from study report
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available
Additional information

Developmental toxicity study:

In a study, test chemical has been investigated for developmental toxicity. Study is summarized below 

The chemical was given to 13 rats per sex per dose level at 0, 750, 1000 and 1250 mg/kg bw/day by oral gave. Male rats were treated two weeks before mating and thereafter for a total of 48 dosing days. Female rats were treated two weeks before mating, during mating, during gestation and during lactation, for a total of approx. 63 dosing days. No morbidity was observed during the study period. No mortality was observed other than two deaths due to gavage error (one male at 750 and one male and 1000 mg/kg). Clinical findings were sporadic and of no biological significance. Body weight changes were restricted to a significant decrease in % body weight change in the recovery group of male rats treated at 1250 mg/kg from day 1 – 22 as compared to the control group. As such, no significant body weight effects were observed for female rats during the study period. Food intake was unaffected by treatment. Hormonal data showed no significant effects on the concentrations of T4 or TSH (male and females), testosterone (males) or oestradiol (females). Oestrous cyclicity was unaffected by treatment and all female rats showed evidence of copulation after the cohabitation/mating period. All females showed precoital intervals of less than 5 days except for four female rats at 750 mg/kg who showed intervals of more than 5 days. Pregnancy rates were 77, 62, 77 and 77% at 0, 750, 1000 and 1250 mg/kg, respectively. No significant effects were observed on gestation length or litter size. Likewise, no significant effects were observed on the number of live births, pup survival, pup weight or sex ratio. Four pups were cannibalized at the 750 mg/kg treatment group. All other pups at 0, 750, 1000 and 1250 mg/kg were normal externally. The internal examination of the pups revealed no abnormalities attributed to the test chemical. Microscopic examination of the pups’ thyroid and parathyroid glands at 0 and 1250 mg/kg revealed no abnormal findings. In the adult rats, no significant effects were observed on the absolute or relative weights of the testes or epididymides. Grossly, one rat treated at 1250 mg/kg showed a mild reduction in the size of the testes. Organ weight data was not collected for the prostate and seminal vesicles with coagulating glands. Gross and microscopic examination of ovaries, uterus, cervix with vagina, testes, epididymides, prostate, seminal vesicle with coagulating glands at 0 and 1250 mg/kg revealed no significant effects attributed to the test chemical. In the testes, mild focal seminiferous tubular degeneration was observed in 2/13, 2/13, 0/13 and 2/13 rats at 0, 750, 1000 and 1250 mg/kg, respectively. Retention of mature sperm (minimal or mild) was observed in 1/13 rats inall treatment groups. Likewise, sloughing of round spermatids/pachytene spermatocytes was observed in 1/13 rats in all treatment groups. NOAEL for the test chemical for systemic and developmental toxicity was established at 1250 mg/kg bw/day in male and female rats following oral gavage exposure before mating, during mating and thereafter for a total of 48 to 63 days.

Justification for classification or non-classification

At present, there is not a sufficient amount of data to make a classification or non-classification for Reproductive toxicity at the tonnage level of 100-1000 T/year. Data from a study similar or equivalent to OECD 414 is needed.