Registration Dossier

Administrative data

Key value for chemical safety assessment

Toxic effect type:
dose-dependent

Effects on fertility

Description of key information

Reproductive toxicity study

Based on the data available from different studies, NOAEL for test material was considered to be in range of 250-500mg/kg bw/day .When male and female rats were treated with test material orally. Thus, comparing this value with the criteria of CLP regulationtest materialisnot likely to classify as reproductive toxicant.

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:
data from handbook or collection of data
Remarks:
Experimental data from various test chemicals
Justification for type of information:
Weight of evidence approach based on the available information from various test chemicals.
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Qualifier:
equivalent or similar to guideline
Guideline:
other: As mentioned below
Principles of method if other than guideline:
WoE report is based on three reproductive toxicity studies on rats
1.Three-generation reproduction toxicity study of test material in Rats.
2&3.Reproductive and developmental toxicity study of test material was performed on wistar rats.
GLP compliance:
not specified
Limit test:
no
Species:
rat
Strain:
other: 1.CD-1 2.Harlan-Wistar 3.Crj: CD (SD) IGS
Sex:
male/female
Details on test animals or test system and environmental conditions:
1.TEST ANIMALS
- Source: Charles River Breeding Laboratories (Wilmington, MA)
- Age at study initiation: (P) x wks; (F1) x wks: P- 80-85 days
- Weight at study initiation: (P) Males: x-x g; Females: x-x g; (F1) Males: x-x g; Females: x-x g: No data available
- Fasting period before study: No data available
- Housing: Animals were housed individually in stainless-steel wire-mesh cages, except during the mating, lactation and post-weaning periods when the females were placed in plastic shoe-box cages containing bedding. Each rat was identified with a metal ear tag. If the tag was lost, the animal was re-tagged and/or toe-clipped.
- Diet (e.g. ad libitum): Basal diet (Purina Rodent Chow from Ralston Purina Co., Inc., St Louis, MO), ad libitum
- Water (e.g. ad libitum): Water, ad libitum
- Acclimation period: No data available

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-21°C
- Humidity (%):40-60%
- Air changes (per hr): No data available
- Photoperiod (hrs dark / hrs light): 12-hr light/dark cycle

3.Details on test animals and env. conditions
TEST ANIMALS
- Source: Charles River Japan Co., Ltd.
- Age at study initiation: 10week old
- Weight at study initiation: 371.7 g (350 to 402 g) for males
231.5 g (195 to 257 g) for female
- Fasting period before study:
- Housing: bracket type metal wire net floor cage.
- Use of restrainers for preventing ingestion (if dermal): yes/no
- Diet (e.g. ad libitum): CRF-
1, Oriental Yeast Industry Co., Ltd
- Water (e.g. ad libitum): drinking water using tap water (Sapporo city)
- Acclimation period:2 week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 to 24 ° C
- Humidity (%):45 to 61%,
- Air changes (per hr): 10 to 15 times / hour
- Photoperiod (hrs dark / hrs light): 12 hours(lit from 8 am to 8 pm)
Route of administration:
other: 1.oral: feed 3.oral: gavage
Type of inhalation exposure (if applicable):
not specified
Vehicle:
other: 1.Basal diet 2.1 w / v% aqueous solution of sodium carmellose
Details on exposure:
1.PREPARATION OF DOSING SOLUTIONS: Test material was mixed with the powdered chow in a twin shell blender to provide theoretical dose levels of 2.5, 25, 75 and 250 mg/kg/day.

DIET PREPARATION
- Rate of preparation of diet (frequency): Weekly
- Mixing appropriate amounts with (Type of food): Basal diet (Purina Rodent Chow from Ralston Purina Co., Inc., St Louis, MO)
- Storage temperature of food: Stored in environmentally controlled room with limited access.

VEHICLE
- Justification for use and choice of vehicle (if other than water): Basal diet
- Concentration in vehicle: 0, 2.5, 25, 75 and 250 mg/kg/day.
- Amount of vehicle (if gavage): No data available
- Lot/batch no. (if required): No data available
- Purity: No data available
3.Details on exposure
PREPARATION OF DOSING SOLUTIONS:
Test material suspended in a 1 w / v% aqueous solution of sodium carmellose
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): 1 w / v% aqueous solution of sodium carmellose
- Concentration in vehicle: 0,250,500,1000 mg/kg bw/day
- Amount of vehicle (if gavage): 10ml/kg bw
- Lot/batch no. (if required):
- Purity:
Details on mating procedure:
1.Details on study schedule
- F1 parental animals not mated until [...] weeks after selected from the F1 litters.
- Selection of parents from F1 generation when pups were [...] days of age.
- Age at mating of the mated animals in the study: [...] weeks
(Explain how study was performed on perents and offspring separately whatever information we have)
F0 geneartion:
F0 animals were meated twice to give F1a and F1b
F1a - on 21-day examined for external abnormalities and killed.
F1b - random selections were made of
ten male and 20 female pups from the control and treated groups of the F1b litters to serve as the second generation (F0 parental rats. The remaining F1b pups were examined for external abnormalities, killed and discarded.

F1 geneartion:
F0 animals were meated twice to give F2a and F2b
F2a - examined for external abnormalities
and killed at the end of the 21-day lactation
period.
F2b – selections were made of ten male and 20 female pups from the control and treated groups to serve as the third generation (F2) parental rats.

F2 geneartion:
F0 animals were meated twice to give F3a , F3b and F3c
F3a – On 21 days old and necropsied and tissues were collected for histopathology.
F3b - F2 animals were remated to give F3a, On 21 days necropsied and tissues were collected for histopathology.
F3c - F2 animals were remated to give F3c. On day 19 of gestation, half of the dams from the control and treated groups were killed.

- M/F ratio per cage: 1:2 ratio
- Length of cohabitation: 15 days
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy Vaginal smears were performed daily until sperm or a copulatory plug was found.
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility. No data available
- Further matings after two unsuccessful attempts: [no / yes (explain)] No data available
- After successful mating each pregnant female was caged (how): Female was placed on corn-cob bedding in an individual plastic shoe-box cage and remained there until it was remated.
- Any other deviations from standard protocol: Each female was rested for a minimum of 10 days after lactation before being mated again.
Each mating was with a different male from the same dosage group.
3.- M/F ratio per cage:1:1
- Length of cohabitation: 14days
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy:the day on which sperm was
confirmed in female vaginal plaque was taken as the 0th 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):
- Any other deviations from standard protocol:
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Prior to initiation of the study, assays were performed to determine the homogeneity and stability of test material in the prepared diets. The concentrations of the test compound in the diets were determined weekly during the first 13 wk of the study and monthly thereafter. All lots of basic feed used in the study were analysed for heavy metals, chlorinated hydrocarbons and aflatoxin. These analyses demonstrated that the basic feed contained acceptably low levels of contaminants, that the diets were prepared properly and that the dietary content of the test material was stable
Duration of treatment / exposure:
Study 1.
224 days
Study 2.Upto 3 generation
Study 3.
Males:46
Females: 14 days before mating and the mating period until mating, and further the mating trial was the period of pregnancy and 3 days of gestation .
Frequency of treatment:
Daily
Details on study schedule:
No data available
Remarks:
Doses / Concentrations:
Study 1.
0, 2.5, 25, 75 and 250 mg/kg/day
Study 2.
0, 5, 50, 150 or 500 mg/kg
Study 3
0,250,500,1000 mg/kg bw/day
No. of animals per sex per dose:
Total: 1410
F0 geneartion:
0 mg/kg bw/day: 20 male, 40 female
2.5 mg/kg bw/day: 10 male, 20 female
25 mg/kg bw/day: 10 male, 20 female
75 mg/kg bw/day: 10 male, 20 female
250 mg/kg bw/day: 10 male, 20 female

F1a geneartion:
0 mg/kg bw/day: 20 male, 40 female
2.5 mg/kg bw/day: 10 male, 20 female
25 mg/kg bw/day: 10 male, 20 female
75 mg/kg bw/day: 10 male, 20 female
250 mg/kg bw/day: 10 male, 20 female

F1b geneartion:
0 mg/kg bw/day: 20 male, 40 female
2.5 mg/kg bw/day: 10 male, 20 female
25 mg/kg bw/day: 10 male, 20 female
75 mg/kg bw/day: 10 male, 20 female
250 mg/kg bw/day: 10 male, 20 female

F2a geneartion:
0 mg/kg bw/day: 20 male, 40 female
2.5 mg/kg bw/day: 10 male, 20 female
25 mg/kg bw/day: 10 male, 20 female
75 mg/kg bw/day: 10 male, 20 female
250 mg/kg bw/day: 10 male, 20 female

F2b geneartion:
0 mg/kg bw/day: 20 male, 40 female
2.5 mg/kg bw/day: 10 male, 20 female
25 mg/kg bw/day: 10 male, 20 female
75 mg/kg bw/day: 10 male, 20 female
250 mg/kg bw/day: 10 male, 20 female

F2c geneartion:
0 mg/kg bw/day: 20 male, 40 female
2.5 mg/kg bw/day: 10 male, 20 female
25 mg/kg bw/day: 10 male, 20 female
75 mg/kg bw/day: 10 male, 20 female
250 mg/kg bw/day: 10 male, 20 female

F3a geneartion:
0 mg/kg bw/day: 20 male, 40 female
2.5 mg/kg bw/day: 10 male, 20 female
25 mg/kg bw/day: 10 male, 20 female
75 mg/kg bw/day: 10 male, 20 female
250 mg/kg bw/day: 10 male, 20 female

F3b geneartion:
0 mg/kg bw/day: 10 male, 20 female
2.5 mg/kg bw/day: 10 male, 20 female
25 mg/kg bw/day: 10 male, 20 female
75 mg/kg bw/day: 10 male, 20 female
250 mg/kg bw/day: 10 male, 20 female
Study 3.
Total:96
0mg/kg bw/day:12 male and 12 female
250 mg/kg bw/day:12 male and 12 female
500 mg/kg bw/day:12 male and 12 female
1000mg/kg bw/day:12 male and 12 female
Control animals:
yes, concurrent vehicle
Details on study design:
No data available
Positive control:
No data available
Parental animals: Observations and examinations:
1.Morbidity and mortality, body weights, food consumption, food efficiency, Foetal development and Histopathology were examined.
3.CAGE SIDE OBSERVATIONS: Yes
- Time schedule:
- For all sexes, behaviors and appearance were observed by visual inspection and palpation at frequencies of once
or more during the test period.
Parental animals observation and examinations
DETAILED CLINICAL OBSERVATIONS: Yes / No / No data
- Time schedule:

BODY WEIGHT: Yes
- Time schedule for examinations: On 1 day of administration (before administration), 2, 5, 7, 10 and 14 days of administration, after every 7 days
(including the administration finish date) for males
for females 0, 1, , 7, 10, 14, 17,
and 20, at 0, 1 and 4 days of nursing, and during the mating period

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Feed intake was measured for males except for the mating period, final administration day and autopsy date, and
for females on the same day as the body weight measurement day except for gestation day 0 and nursing 0 day,
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes / No / No data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes / No / No data

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

Oestrous cyclicity (parental animals):
yes observed
Sperm parameters (parental animals):
No data available
Litter observations:
1.Morbidity and mortality, body weights, food consumption and food efficiency of F1 and F2 geneation were examined.
2.On the 4th day of nursing, euthanasia lethally using the carbon dioxide inhalation method, and visually observe all the organs and tissues macroscopically . For dead and abnormal finding sites, whole body was fixed with 10% neutral buffered formalin and stored.
Postmortem examinations (parental animals):
1&3.Gross pathology and Histopathology were examined.
Postmortem examinations (offspring):
1.Foetal development and Histopathology were examined.
3.On the 4th day of nursing, euthanasia lethally using the carbon dioxide inhalation method, and visually observe all the organs and tissues macroscopically . For dead and abnormal finding sites, whole body was fixed with 10% neutral buffered formalin and stored.
Statistics:
1.Fertility indices were compared using the chisquare test criterion with Yates' correction for 2 × 2 contingency tables (Steel & Torrie, 1960) to judge the significance of differences. Gestation, 4-day, 14-day and 21-day survival indices were compared by the rank sum tests described by Snedecor & Cochran (1967) and Weil (1970) to judge the significance of differences. The numbers of pups born alive were compared by analysis of variance and the t test, as described by Steel & Torrie (1960), using the multiple comparison tables of Dunnett (1964) to judge the significance of differences.
3.Multiple sample χ ^ 2- test was performed on findings that showed one level of positive grade among sexual cycle, mating rate, conception rate, childbirth rate and nursing rate, and histopathological examination results, and in case of significant 2 Sample c 2 - Assay was performed. In addition, Fisher's direct probability test method was used when these tests failed. For other observations and findings showing positive grade of 2 or more out of the results of histopathological examination, after equality dispersal test of Bartlett, analyzed by one way analysis of variance or Kruskal-Wallis method, significant , The comparison group and the administration group were compared by Dunnett's test method or Mann-Whitney U-test method. For the test with the control group, the significance level was set at 5%.
Reproductive indices:
1.Fertility indices, gestation and lactation indices were examined.
Offspring viability indices:
Yes, viability on day 4, 14 and 21 were examined.
Clinical signs:
no effects observed
Description (incidence and severity):
1.When treated with 250 mg/kg body weight/day, Slightly bluish-coloured fur and Bluishgreen- coloured faeces were observed in treated rats as compared to control. When treated with 75 mg/kg body weight/day, Bluishgreen- coloured faeces were observed in treated rats as compared to control. Occasional ocular or nasal porphyrin discharge, soft tools, respiratory congestion and slight alopecia were observed in treated rats which were in similar frequency with control.
3.In males, hair removal and crusts were observed in one patient in the 250 mg / kg group from 37 days to the autopsy date, but were accidental, not seen in other groups.
In females, no abnormality was observed in any of pre-pregnancy administration period, pregnancy period and nursing period.

Dermal irritation (if dermal study):
not specified
Mortality:
no mortality observed
Description (incidence):
1&3.No effect on survival of treated rats were observed as compared to control.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
1.No effect on body weight of treated rats was observed as compared to control.
3.No significant difference was observed between treated group and control group in any of period of male administration, female pre-pregnancy administration period, pregnancy period and nursing period.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
1.Although variations in food consumption were recorded for all groups, no marked or consistent differences were observed between control and treated groups.
During the mating period, the diets fed were prepared on the basis of the females' body weights and food consumption. Therefore, the amount of colouring consumed by male rats during this period was slightly lower than the required dosage level.
3.In males, low food intake was observed on day 46 of administration in the 500 mg / kg group, but it was a transient change with no dose correlation.
In females, there was no significant difference in treated administration group in each pregnancy administration period, pregnancy period and nursing period compared with the control group in each group.

Food efficiency:
no effects observed
Description (incidence and severity):
1.No effect on food efficiency of treated rast were observed as compared to control.
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
3.Reduction of activated partial thromboplastin time was seen in the 500 and 1000 mg / kg group of male, and a significant difference was observed as compared with the control group.
Clinical biochemistry findings:
not specified
Urinalysis findings:
effects observed, treatment-related
Description (incidence and severity):
3.A decrease in pH (range: 6.0 to 8.0 in the range of 500 and 1000 mg / kg group, control group; 8.0 to 8.5) was observed in the male 500 and 1000 mg / kg group, and a significant difference was observed as compared with the control group
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
3.Histopathological examination revealed eosinophilic corpuscle of the proximal tubular epithelium in the kidney in males in 1 to 3 cases in the 250, 500 and 1000 mg / kg groups, but in comparison with the control group No significant difference was observed and it was not considered to be an effect of test material administration no dose-dependent increase was observed in the collection of lung foam cells and the number of occurrences of lymphocyte infiltration of the prostate, and the other findings were also changed only in one case, which is not related to test material administration. Histological examination of females showed only one case in the 1000 mg / kg group, but peripallal fatty liver in the liver, fatty degeneration of the proximal tubular epithelium in the kidney and atrophy of the thymus were observed in the kidney
Histopathological findings: neoplastic:
not specified
Other effects:
no effects observed
Reproductive function: oestrous cycle:
not specified
Description (incidence and severity):
In the female sexual cycle examination, consecutive consecutive periods of estrous period were observed in each case in the 500 and 1000 mg / kg groups and in one case of irregular estrus in the 500 mg / kg group during the administration period during the mating. In all these cases, mating was established, but inferiority was found in 2 groups of 1000 mg / kg group, among which one set of females was an example in which the estrus pause period continued.
Reproductive function: sperm measures:
not specified
Reproductive performance:
no effects observed
Description (incidence and severity):
1&2.no indications of any influence on reproductive parameters.
3.No abnormalities were observed in the ovaries, testis and epididymis in the infertility cases of this 1000 mg / kg group and necropsy and histopathological examination of mating male. In addition, as compared with the control group, no significant difference was observed in treated groups on the number of days before copulation, mating rate and conception rate.

No significant difference was observed between treated group for gestation period, nursing rate, pregnancy luteum count, implantation number, implantation rate, number of babies, birth rate, number of surviving babies at birth confirmation and fertility rate. The high delivery rate was found in the 1000 mg / kg group, as a result of the fact that the number of children in the same group is larger than that in the control group.
One patient in the 500 mg / kg group did not deliver by 26th day of pregnancy, lesion expansion of the left and right uterus, uterine cervix and vagina was observed at the autopsy of this example, and 5 implantation marks were observed However, surviving fetuses were not observed.
Mortality: No effect on survival of treated rats were observed as compared to control.

Clinical signs: When treated with 250 mg/kg body weight/day, Slightly bluish-coloured fur and Bluishgreen- coloured faeces were observed in treated rats as compared to control.

When treated with 75 mg/kg body weight/day, Bluishgreen-
coloured faeces were observed in treated rats as compared to control.

Occasional ocular or nasal porphyrin discharge, soft tools, respiratory congestion and slight alopecia were observed in treated rats which were in similar frequency with control.

Body weight: No effect on body weight of treated rats was observed as compared to control.

Food consumption: Although variations in food consumption were recorded for all groups, no marked or consistent differences were observed between control and treated groups.

During the mating period, the diets fed were prepared on the basis of the females' body weights and food consumption. Therefore, the amount of colouring consumed by male rats during this period was slightly lower than the required dosage level.

Test substance intake: During the weaning period, the pups were not prevented from eating the food provided for the dams; the consumption of colouring by female rats during the gestation and weaning periods therefore appeared to be slightly higher than anticipated.

Reproductive function: estrous cycle
F2 parentes: No effect on live and dead foetuses, resorptions and corpora lutea of treated rats were observed as compared to control.

Reproductive function: sperm measures: No data available

Reproductive performance
F1 parentes: When treated with 2.5 and 250 mg/kg body weight/day, significant decrase in fertility indices of treated female rats were observed as compared to control.

Because of the lower fertility indices observed at the 2.5 mg/kg bw/day dosage group in the F 2 litters, the study was extended to the F3b litter.

No effect on F0 and F3 generation fertility indices were observed as compared to control.

The changes were considered to represent variation rather than compound-related effects.

Organ weights No effect on organ weight of treated rats were oberved as compared to control.

Gross pathology: No effect on Gross pathology of treated rats were oberved as compared to control.

Histopathology: No data available

other findings
Food efficiency: No effect on food efficiency of treated rast were observed as compared to control.

Dose descriptor:
NOAEL
Effect level:
> 250 - <= 500 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No adverse effect on survival, body weights, food consumption, food efficiency, Estrous cyclicity, Foetal development, organ weights and gross pathology
Remarks on result:
other: No reproductive toxic effects observed
Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Treatment related:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified
Clinical signs:
no effects observed
Description (incidence and severity):
1.Slightly bluishcoloured fur was noted inpups given 250mg/kg/day and bluish-greencoloured faeces were produced by pups given 75 or 250 mg/kg/day. No other changes considered to be compound related were noted in general behaviour or appearance. Changes noted occasionally in control or treated pups included alopecia, respiratory congestion and runting.
Dermal irritation (if dermal study):
not specified
Mortality / viability:
no mortality observed
Description (incidence and severity):
1. No effect on survival of F1 pups were observed as compared to control.
2.There was no significant difference in the survival rate of the newborn on the 4th day compared to the control group and treated group.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
1.The average body weights of the pups were similar in control and treated groups. Although the average number of pups born per litter varied among the groups, no dose relationship was apparent.
2.Compared with the control group, the low value of body weight of 4 days of nursing in males of the 500 mg / kg group, the 4 days of nursing of body weight gain, the weight gain and the body weight gain rate were low in females as compared with the control group. Even in the 1000 mg / kg group, low values of weight of body weight of 4 days of nursing were observed in male, 4 days of body weight in nursing and low value of body
weight gain were observed in females
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:
no effects observed
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
There were no compound-related effects on organ Weights.
Gross pathological findings:
no effects observed
Description (incidence and severity):
1.No compound-related gross pathological change in any of the rats that were killed and necropsied
3.Deficits in the tail tips of the control group observed in general condition observation and one in each of the 1000 mg / kg group, and abdominal or cervical trauma in 1 case of each sex of 1000 mg / kg group were observed , No other abnormalities were found in any of the dead cases and autopsy of newborn sacrificed on the 4th day of nursing in any animals.
Histopathological findings:
no effects observed
Other effects:
not specified
Behaviour (functional findings):
not specified
Developmental immunotoxicity:
not specified
Mortality: No effect on survival of F1 and F2 pups were observed as compared to control.

Clinical signs: When treated with 250 mg/kg body weight/day, Slightly bluish-coloured fur and Bluishgreen- coloured faeces were observed in treated pups as compared to control.

When treated with 75 mg/kg body weight/day, Bluishgreen-
coloured faeces were observed in treated pups as compared to control.

Respiratory congestion and runting were observed in treated rats which were in similar frequency with control.

Body weight:
F1 and F2 pups:
No effect on body weight of treated pups was observed as compared to control.

Food consumption:
F1 and F2 pups:
No effect on food consumption of treated pups was observed as compared to control.

Organ weights: No effect on organ weight of treated pups were oberved as compared to control.

Gross pathology: No effect on Gross pathology of treated pups were oberved as compared to control.

Histopathology: No gross anatomical abnormalities were observed in fetouse of treated rats as compared to control.
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
> 250 - <= 500 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No adverse effect on survival, body weights, food consumption, food efficiency, Estrous cyclicity, Foetal development, organ weights, Gross pathology and Histopathology
Remarks on result:
other: No developmental toxic effects was observed
Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Treatment related:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified
Clinical signs:
no effects observed
Description (incidence and severity):
Slightly bluishcoloured fur was noted inpups given 250mg/kg/day and bluish-greencoloured faeces were produced by pups given 75 or 250 mg/kg/day. No other changes considered to be compound related were noted in general behaviour or appearance. Changes noted occasionally in control or treated pups included alopecia, respiratory congestion and runting.
Dermal irritation (if dermal study):
not specified
Mortality / viability:
no mortality observed
Description (incidence and severity):
No effect on survival of F2 pups were observed as compared to control.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
The average body weights of the pups were similar in control and treated groups. Although the average number of pups born per litter varied among the groups, no dose relationship was apparent.
Food consumption and compound intake (if feeding study):
no effects observed
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
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
no effects observed
Description (incidence and severity):
No compound-related gross pathological change in any of the rats thatwerekilled and necropsied
Histopathological findings:
no effects observed
Description (incidence and severity):
No compound-related microscopic pathological lesions in any of the rats that were killed and necropsied
Other effects:
no effects observed
Behaviour (functional findings):
not specified
Developmental immunotoxicity:
not specified
Dose descriptor:
NOAEL
Generation:
F2
Effect level:
250 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No adverse effect on survival, body weights, food consumption, food efficiency, Estrous cyclicity, Foetal development, organ weights, Gross pathology and Histopathology
Remarks on result:
other: No developmental toxic effects observed
Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Treatment related:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified
Reproductive effects observed:
not specified
Treatment related:
not specified
Relation to other toxic effects:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified
Conclusions:
NOAEL was considered to be in range of 250 - 500 mg/kg body weight/day for F0, F1 generation, when male and female rats were treated with test material orally.
Executive summary:

Data available from different studies were reviewed to determine the reproductive toxicity of test material .The studies are as mentioned below:

Study 1

In a three generation reproduction study, groups of ten males and twenty female Charles River CD rats were fed with test material  at dietary levels providing intakes of 0, 2.5, 25, 75 and 250 mg/kg bw/day. Two females were placed in a male's cage for the entire mating period (15 days). Vaginal smears were performed daily until sperm or a copulatory plug was found; this was designated as day 0 of pregnancy. At the end of the mating period, each

female was placed on corn-cob bedding in an individual plastic shoe-box cage and remained there until it was remated. Each female was rested for a minimum of 10 days after lactation before being mated again. Each mating was with a different male from the same dosage group.

The control and treated F0 rats were maintained on their respective diets for 2 wk prior to the first mating period. They were then mated, when approximately 100 days old. The rats (F0) were mated twice. The pups (F1a) from the first mating were examined for external abnormalities and killed at the end of the 21-day lactation period. After the second mating, random selections were made of ten male and 20 female pups from the control and treated groups of the F1b litters to serve as the second generation (F1) parental rats. The remaining F1b pups were examined for external abnormalities, killed and discarded. The F0 parent rats were also killed at this time.

After weaning, the F1b pups selected for the second generation continued to be fed on their control or appropriately treated diet for 80 days and were then mated. The pups from the first mating (F2a) were examined for external abnormalities and killed at the end of the 21-day lactation period. From the second mating (F2b), selections were made of ten male and 20 female pups from the control and treated groups to serve as the third generation (F2) parental rats. The remaining pups (F2b) were examined for external abnormalities, killed and discarded. Following the third mating, one-half of the mated females were killed on day 19 of gestation and the uteri and ovaries were examined. The remaining females were allowed to deliver. After weaning, the F2c pups were examined for external abnormalities, killed and discarded. The F1 parent rats were then killed. Five male and five female rats from the control and each of the treated groups were necropsied and their tissues were collected for histopathology.

After weaning, the F2b pups selected for the third generation continued to be fed on their control or appropriately treated diet for 80 days and were then mated. Five male and five female pups (F3a) from the control and each of the treated groups were killed when 21 days old and necropsied, and tissues were collected for histopathology. The parent rats (F2) were then re-mated and the pups from the second mating (F3b) were examined for external abnormalities and killed at the end of the 21-day lactation period. The F 2 parent rats were then re-mated for the third mating (F3c). On day 19 of gestation, half of the dams from the control and

treated groups were killed by chloroform anesthesia. Uteri were examined for any abnormal conditions and the presence of live and dead foetuses and of resorptions were recorded. All ovaries were inspected and the corpora lutea were counted. Remaining F2 parent rats were then killed and discarded.

The rats were observed twice daily for changes in behaviour or appearance and for morbidity and mortality. Individual body weights and food consumption were recorded weekly. Specific parameters for the reproductive phase of this study included observations of fertility, litter size, numbers of male and female pups, viability of the newborn, survival of pups to weaning and growth of the pups.All stillborn offspring and any progeny that died during the study were examined either by skeletal clearing or by necropsy. Body weights of pups aged 4 days (before and after reduction of litters to ten pups) and aged 14 days were obtained by weighing as litters. The pups were weighed individually at 21 days of age. The thyroid, adrenals, lung. heart, spleen, stomach, jejunum, ileum, colon, liver, kidneys, urinary bladder, testes or ovaries and uterus from five male and five female rats from the F1 and F3, generations of the control and 250-mg/kg/day groups were embedded in paraffin, sectioned, stained with haematoxylin and eosin and examined microscopically.

 

Fur and faeces were bluish-coloured in 75 mg/kg bw/day and 250 mg/kg bw/day groups. Gestation, viability and lactation indices of all litters from exposed animals did not differ from controls. Fertility indices were statistically significantly lower for F2female rats in the 2.5 and 25 mg/kg bw/day groups only and consequently considered as not being dose-related. Fertility indices were also reduced in F2b and F2cgroup of male rats. No statistically significant changes in the fertility index were observed in the F3generation. As effects on fertility indices in the F2generation were not dose-related and effects were not identified in the F1and F3generation this effect was not considered to be compound-related. Examination of the ovaries and uteri of F1dams killed on gestation day (GD) 19 revealed no gross anatomical abnormalities. No unusual changes were observed in the stillborn pups or in pups that died during the study. No compound-related gross or microscopic pathological lesions were noted in any of the F1or F3arats that were sacrificed and necropsied. Finally, no compound-related organ-weight variations were recorded in the F1rats. ThereforeNOAEL was considered to be250 mg /kg bw/day for test material  inCharles RiverCD male and female rats by oral administration (feed) in 3 generation study.(F0,F1 andF2 generation)

 

 Study 2.

Reproductive and development toxicity study oftestmaterial was performed on male and femaleHarlan-Wistar rats. Test material administered in diet for up to 3 generation. 10 male and 20 females were used. Asno indications of any influence on

maternal weight gain,reproductive parameters andNo developmental toxic effects were seen up to the highest dose of 500 mg/kg bw, Hence No Observed Adverse Effect Level (NOAEL) for maternal toxicity was considered to be 500 mg/kg/day.When male and femalewistar rats were treated withtest materialorally up to 3 generation.

Study 3

The reproductive and developmental toxicity of test material was performed on male and female rats. The test material suspended in 1 w / v% aqueous solution of sodium carmellose in dose concentration0, 250, 500, 1000 mg/kg bw/day and administered orally by gavage. The dose concentration were selected on the bases on dose finding study (5, 70 and 1000 mg / kg).Exposure to test material for the males, 46 days including the mating period and for the females for 14 days before mating and the mating period until mating, and further the mating trial was the period of pregnancy and 3 days of gestation. For males and females on 14th day of administration, they were allowed to live together in the same group within 1 to 1 (random combination) from the evening for only 14 days and the day on which sperm was confirmed in female vaginal plaque was taken as the 0th gestation. For all sexes, behaviours and appearance were observed by visual inspection and palpation at frequencies of once or more during the test period. For all female mated females, from the 21st day of pregnancy to the end of calving, the status of parturition, nursing behaviour, the number of total births, the number of surviving children and the number of dead children, the sex and the outer table of the infant were observed.

In males, hair removal and crusts were observed in one patient in the 250 mg / kg group from 37 days to the autopsy date, but were accidental, not seen in other groups. In females, no abnormality was observed in any of pre-pregnancy administration period, pregnancy period and nursing period.No significant difference was observed between treated group and control group in any of period of male administration, female pre-pregnancy administration period, pregnancy period and nursing period.In males, low food intake was observed on day 46 of administration in the 500 mg / kg group, but it was a transient change with no dose correlation. In females, there was no significant difference in treated administration group in each pregnancy administration period, pregnancy period and nursing period compared with the control group in each group.

In reproductive tests, male and female mating ratios, female sexual cycles and conception rates, test material administration in the necropsy, weight and histopathological examination of reproductive organs (testis, epididymis and ovaries) and endocrine organs (pituitary, adrenal) No effect was observed. On the other hand, pathological examination of the infertile reproductive organs of 2 cases observed in the 1000 mg / kg group showed no abnormality and no influence by test material administration was observed.

There was no effect of test material administration on maternal necropsy, pregnancy period, number of corpus luteums, implantation number, implantation rate, birth rate, delivery rate, number of births and number of surviving children and birth rate at birth confirmation. However, in one case of the 1000 mg / kg group, among 9 males and 8 females, 1 male and 1 female died on 2 nursing, 2 male on 3 nursing and 1 male female on nursery 4.Three cases were unknown, and the weight of surviving newborn infants was decreasing. In this mother animals, body weight loss and low food intake were also observed during the nursing period, and histopathological examination revealed peripallal fatty liver in the liver, fatty degeneration of the proximal tubular epithelium in the kidney and thymic Atrophy was observed. Therefore, although only one case appeared, the possibility that test material administration might have affected the mother's ability to nurture was considered.

In addition, in one case of 500 mg / kg group where delivery was not observed until 26th day of pregnancy, laparotomy of left and right uterus and cervix was observed at necropsy. In the same example, five implantation marks were seen by observation with the naked eye, both of which are considered to be early embryonic deaths, and the relation with test material administration was observed. survival were observed, in neonates weighing transition observed low values of body weight nursing 4 days in male and female 500 and 1000 mg / kg group, In addition, in the 500 mg / kg females, the body weight gain and the body weight gain rate were low, and even in females of the 1000 mg / kg group, the body weight gain was low. But neonatal weight during delivery was not changed in each group treated with test material. Therefore, a decrease or milk migration nursing ability of dams by test material administration was believed to have affected the newborn weighing 500 and 1000 mg / kg group, including one example of 1000 mg / kg group described above, The mechanism could not be clarifiedunder this test condition. Hence The no effect level (NOEL) for reproduction of parent animals by repetitive oral administration of test material in this study was considered to be 1,000 mg / kg / day in male, 250 mg / kg / day for females, and 250 mg / kg / day for no effect on neonatal development (NOEL).

 

Based on the data available from different studies,Test material did not showed reproductive toxicityat dose concentration 250 mg/kg bw/day by oral route.Hence the test chemical is not likely to classify as a reproductive toxicant as per the criteria mentioned in CLP regulation.

 

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

Toxicity to reproduction

Data available from different studies were reviewed to determine the reproductive toxicity of test material.The studies are as mentioned below:

Study 1

In a three generation reproduction study, groups of ten males and twenty female Charles River CD rats were fed with test material  at dietary levels providing intakes of 0, 2.5, 25, 75 and 250 mg/kg bw/day. Two females were placed in a male's cage for the entire mating period (15 days). Vaginal smears were performed daily until sperm or a copulatory plug was found; this was designated as day 0 of pregnancy. At the end of the mating period, each

female was placed on corn-cob bedding in an individual plastic shoe-box cage and remained there until it was remated. Each female was rested for a minimum of 10 days after lactation before being mated again. Each mating was with a different male from the same dosage group.

The control and treated F0 rats were maintained on their respective diets for 2 wk prior to the first mating period. They were then mated, when approximately 100 days old. The rats (F0) were mated twice. The pups (F1a) from the first mating were examined for external abnormalities and killed at the end of the 21-day lactation period. After the second mating, random selections were made of ten male and 20 female pups from the control and treated groups of the F1b litters to serve as the second generation (F1) parental rats. The remaining F1b pups were examined for external abnormalities, killed and discarded. The F0 parent rats were also killed at this time.

After weaning, the F1b pups selected for the second generation continued to be fed on their control or appropriately treated diet for 80 days and were then mated. The pups from the first mating (F2a) were examined for external abnormalities and killed at the end of the 21-day lactation period. From the second mating (F2b), selections were made of ten male and 20 female pups from the control and treated groups to serve as the third generation (F2) parental rats. The remaining pups (F2b) were examined for external abnormalities, killed and discarded. Following the third mating, one-half of the mated females were killed on day 19 of gestation and the uteri and ovaries were examined. The remaining females were allowed to deliver. After weaning, the F2c pups were examined for external abnormalities, killed and discarded. The F1 parent rats were then killed. Five male and five female rats from the control and each of the treated groups were necropsied and their tissues were collected for histopathology.

After weaning, the F2b pups selected for the third generation continued to be fed on their control or appropriately treated diet for 80 days and were then mated. Five male and five female pups (F3a) from the control and each of the treated groups were killed when 21 days old and necropsied, and tissues were collected for histopathology. The parent rats (F2) were then re-mated and the pups from the second mating (F3b) were examined for external abnormalities and killed at the end of the 21-day lactation period. The F 2 parent rats were then re-mated for the third mating (F3c). On day 19 of gestation, half of the dams from the control and

treated groups were killed by chloroform anesthesia. Uteri were examined for any abnormal conditions and the presence of live and dead foetuses and of resorptions were recorded. All ovaries were inspected and the corpora lutea were counted. Remaining F2 parent rats were then killed and discarded.

The rats were observed twice daily for changes in behaviour or appearance and for morbidity and mortality. Individual body weights and food consumption were recorded weekly. Specific parameters for the reproductive phase of this study included observations of fertility, litter size, numbers of male and female pups, viability of the newborn, survival of pups to weaning and growth of the pups.All stillborn offspring and any progeny that died during the study were examined either by skeletal clearing or by necropsy. Body weights of pups aged 4 days (before and after reduction of litters to ten pups) and aged 14 days were obtained by weighing as litters. The pups were weighed individually at 21 days of age. The thyroid, adrenals, lung. heart, spleen, stomach, jejunum, ileum, colon, liver, kidneys, urinary bladder, testes or ovaries and uterus from five male and five female rats from the F1 and F3, generations of the control and 250-mg/kg/day groups were embedded in paraffin, sectioned, stained with haematoxylin and eosin and examined microscopically.

 

Fur and faeces were bluish-coloured in 75 mg/kg bw/day and 250 mg/kg bw/day groups. Gestation, viability and lactation indices of all litters from exposed animals did not differ from controls. Fertility indices were statistically significantly lower for F2female rats in the 2.5 and 25 mg/kg bw/day groups only and consequently considered as not being dose-related. Fertility indices were also reduced in F2b and F2cgroup of male rats. No statistically significant changes in the fertility index were observed in the F3generation. As effects on fertility indices in the F2generation were not dose-related and effects were not identified in the F1and F3generation this effect was not considered to be compound-related. Examination of the ovaries and uteri of F1dams killed on gestation day (GD) 19 revealed no gross anatomical abnormalities. No unusual changes were observed in the stillborn pups or in pups that died during the study. No compound-related gross or microscopic pathological lesions were noted in any of the F1or F3arats that were sacrificed and necropsied. Finally, no compound-related organ-weight variations were recorded in the F1rats. ThereforeNOAEL was considered to be250 mg /kg bw/day for test material  inCharles RiverCD male and female rats by oral administration (feed) in 3 generation study.(F0,F1 andF2 generation)

 

 Study 2.

Reproductive and development toxicity study oftestmaterial was performed on male and femaleHarlan-Wistar rats. Test material administered in diet for up to 3 generation. 10 male and 20 females were used. Asno indications of any influence on

maternal weight gain,reproductive parameters andNo developmental toxic effects were seen up to the highest dose of 500 mg/kg bw, Hence No Observed Adverse Effect Level (NOAEL) for maternal toxicity was considered to be 500 mg/kg/day.When male and femalewistar rats were treated withtest materialorally up to 3 generation.

Study 3

The reproductive and developmental toxicity of test material was performed on male and female rats. The test material suspended in 1 w / v% aqueous solution of sodium carmellose in dose concentration0, 250, 500, 1000 mg/kg bw/day and administered orally by gavage. The dose concentration were selected on the bases on dose finding study (5, 70 and 1000 mg / kg).Exposure to test material for the males, 46 days including the mating period and for the females for 14 days before mating and the mating period until mating, and further the mating trial was the period of pregnancy and 3 days of gestation. For males and females on 14th day of administration, they were allowed to live together in the same group within 1 to 1 (random combination) from the evening for only 14 days and the day on which sperm was confirmed in female vaginal plaque was taken as the 0th gestation. For all sexes, behaviours and appearance were observed by visual inspection and palpation at frequencies of once or more during the test period. For all female mated females, from the 21st day of pregnancy to the end of calving, the status of parturition, nursing behaviour, the number of total births, the number of surviving children and the number of dead children, the sex and the outer table of the infant were observed.

In males, hair removal and crusts were observed in one patient in the 250 mg / kg group from 37 days to the autopsy date, but were accidental, not seen in other groups. In females, no abnormality was observed in any of pre-pregnancy administration period, pregnancy period and nursing period.No significant difference was observed between treated group and control group in any of period of male administration, female pre-pregnancy administration period, pregnancy period and nursing period.In males, low food intake was observed on day 46 of administration in the 500 mg / kg group, but it was a transient change with no dose correlation. In females, there was no significant difference in treated administration group in each pregnancy administration period, pregnancy period and nursing period compared with the control group in each group.

In reproductive tests, male and female mating ratios, female sexual cycles and conception rates, test material administration in the necropsy, weight and histopathological examination of reproductive organs (testis, epididymis and ovaries) and endocrine organs (pituitary, adrenal) No effect was observed. On the other hand, pathological examination of the infertile reproductive organs of 2 cases observed in the 1000 mg / kg group showed no abnormality and no influence by test material administration was observed.

There was no effect of test material administration on maternal necropsy, pregnancy period, number of corpus luteums, implantation number, implantation rate, birth rate, delivery rate, number of births and number of surviving children and birth rate at birth confirmation. However, in one case of the 1000 mg / kg group, among 9 males and 8 females, 1 male and 1 female died on 2 nursing, 2 male on 3 nursing and 1 male female on nursery 4.Three cases were unknown, and the weight of surviving newborn infants was decreasing. In this mother animals, body weight loss and low food intake were also observed during the nursing period, and histopathological examination revealed peripallal fatty liver in the liver, fatty degeneration of the proximal tubular epithelium in the kidney and thymic Atrophy was observed. Therefore, although only one case appeared, the possibility that test material administration might have affected the mother's ability to nurture was considered.

In addition, in one case of 500 mg / kg group where delivery was not observed until 26th day of pregnancy, laparotomy of left and right uterus and cervix was observed at necropsy. In the same example, five implantation marks were seen by observation with the naked eye, both of which are considered to be early embryonic deaths, and the relation with test material administration was observed. survival were observed, in neonates weighing transition observed low values of body weight nursing 4 days in male and female 500 and 1000 mg / kg group, In addition, in the 500 mg / kg females, the body weight gain and the body weight gain rate were low, and even in females of the 1000 mg / kg group, the body weight gain was low. But neonatal weight during delivery was not changed in each group treated with test material. Therefore, a decrease or milk migration nursing ability of dams by test material administration was believed to have affected the newborn weighing 500 and 1000 mg / kg group, including one example of 1000 mg / kg group described above, The mechanism could not be clarifiedunder this test condition. Hence The no effect level (NOEL) for reproduction of parent animals by repetitive oral administration of test material in this study was considered to be 1,000 mg / kg / day in male, 250 mg / kg / day for females, and 250 mg / kg / day for no effect on neonatal development (NOEL).

 

Based on the data available from different studies,Test material did not showedreproductive toxicityat dose concentration 250mg/kg bw/day by oral route.Hence the test chemical is not likely to classify as a reproductive toxicant as per the criteria mentioned in CLP regulation.

 

Effects on developmental toxicity

Description of key information

Developmental toxicity study

Based on the various studies available for the test chemical were reviewed to determine the developmental toxicity, NOAELfor test chemical was considered to be 250/kg bw/day .When rats were treated with test chemical orally. Thus, comparing this value with the criteria ofCLP regulation test chemical is not likely to classify as reproductive and developmental toxicant.

Link to relevant study records
Reference
Endpoint:
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:
data from handbook or collection of data
Remarks:
Experimental data from various test chemicals
Justification for type of information:
Data is summarized based on the available information from various test chemicals.
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Qualifier:
equivalent or similar to guideline
Guideline:
other: As mentioned below
Principles of method if other than guideline:
WoE report is based on two developmental toxicity studies on rats
Repeated oral administration toxicity / reproductive developmental toxicity combined test using test material in rat.
GLP compliance:
not specified
Limit test:
no
Species:
rat
Strain:
other: 1.Crj: CD (SD) IGS 2.CD-1
Details on test animals or test system and environmental conditions:
1.Details on test animals and env. conditions
TEST ANIMALS
- Source: Charles River Japan Co., Ltd.
- Age at study initiation: 10week old
- Weight at study initiation: 371.7 g (350 to 402 g) for males
231.5 g (195 to 257 g) for female
- Fasting period before study:
- Housing: bracket type metal wire net floor cage.
- Use of restrainers for preventing ingestion (if dermal): yes/no
- Diet (e.g. ad libitum): CRF-1, Oriental Yeast Industry Co., Ltd, ad libitum
- Water (e.g. ad libitum): drinking water using tap water (Sapporo city) ad libitum
- Acclimation period:2 week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 to 24 ° C
- Humidity (%):45 to 61%,
- Air changes (per hr): 10 to 15 times / hour
- Photoperiod (hrs dark / hrs light): 12 hours (lit from 8 am to 8 pm)
2.TEST ANIMALS
- Source: Charles River Breeding Laboratories
(Wilmington, MA, USA)
- Age at study initiation: 90 days old
- Weight at study initiation: 250 – 275 g female
- Fasting period before study: No data available
- Housing: Females were housed individually (after mating) in suspended stainless-steel wire-mesh cages. Each rat was identified with a metal ear tag.
- Diet (e.g. ad libitum): Food (Purina Rodent Chow from Ralston Purina Co., Inc., St Louis, MO), ad libitum
- Water (e.g. ad libitum): Water, ad libitum
- Acclimation period: 2 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-21°C
- Humidity (%):40-60%
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light): 12-hr light/dark cycle

IN-LIFE DATES: From: To: No data available

Route of administration:
oral: gavage
Vehicle:
other: 1.1 w / v% aqueous solution of sodium carmellose 2. 0.5% Methocel
Details on exposure:
1.Details on exposure
PREPARATION OF DOSING SOLUTIONS:
Test material suspended in a 1 w / v% aqueous solution of sodium carmellose
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): 1 w / v% aqueous solution of sodium carmellose
- Concentration in vehicle: 0,250,500,1000 mg/kg bw/day
- Amount of vehicle (if gavage): 10ml/kg bw
- Lot/batch no. (if required):
- Purity:
2.PREPARATION OF DOSING SOLUTIONS: Test chemical was dissolved in 0.5% Methocel

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): 0.5% Methocel
- Concentration in vehicle: 0, 25, 75 and 250 mg/kg/day
- Amount of vehicle (if gavage): 1.0 ml/body weight
- Lot/batch no. (if required): No data available
- Purity: No data available
Analytical verification of doses or concentrations:
not specified
Details on mating procedure:
1.- M/F ratio per cage:1:1
- Length of cohabitation:14 days
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy:the day on which sperm was
confirmed in female vaginal plaque was taken as the 0th gestation
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility.:No data available
- Further matings after two unsuccessful attempts: [no / yes (explain)]No data available
- After successful mating each pregnant female was caged (how):No data available
2.- M/F ratio per cage: 1: 2 ratio
- Length of cohabitation: Until evidence of copulation was found
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy The day of evidence of copulation were considered to be day 0 of pregnancy.
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility. No data available
- Further matings after two unsuccessful attempts: [no / yes (explain)]: No data available
- After successful mating each pregnant female was caged (how): Individually
- Any other deviations from standard protocol: No data available

Duration of treatment / exposure:
1.Males:46
Females: 14 days before mating and the mating period until mating, and further the mating trial was the period of pregnancy and 3 days of gestation.
2.10 days
Frequency of treatment:
Daily
Duration of test:
1.Approx 54 days
2. 20 days
Remarks:
Study 1
0,250,500,1000 mg/kg bw/day
Study 2.
0,25,75,250mg/kg /day
No. of animals per sex per dose:
Study 1
Total:96
0mg/kg bw/day:12 male and 12 female
250 mg/kg bw/day:12 male and 12 female
500 mg/kg bw/day:12 male and 12 female
1000mg/kg bw/day:12 male and 12 female
Study 2.
Total: 80
0 mg/kg/day: 20 female
25 mg/kg/day: 20 female
75 mg/kg/day: 20 female
250 mg/kg/day: 20 female
Control animals:
yes, concurrent vehicle
Details on study design:
No data available
Maternal examinations:
study 1&2
CAGE SIDE OBSERVATIONS: Yes
- Time schedule:
- For all sexes, behaviours and appearance were observed by visual inspection and palpation at frequencies of once or more during the test period.
Parental animals observation and examinations
DETAILED CLINICAL OBSERVATIONS: Yes / No / No data
- Time schedule:

BODY WEIGHT: Yes
- Time schedule for examinations: On 1 day of administration (before administration), 2, 5, 7, 10 and 14 days of administration, after every 7 days (including the administration finish date) for males, for females 0, 1, , 7, 10, 14, 17, and 20, at 0, 1 and 4 days of nursing, and during the mating period

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Feed intake was measured for males except for the mating period, final administration day and autopsy date, and
for females on the same day as the body weight measurement day except for gestation day 0 and nursing 0 day,
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes / No / No data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes / No / No data

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes / No / No data
- Time schedule for examinations:
Ovaries and uterine content:
study 1&2
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Other:
Fetal examinations:
study 1&2
- External examinations: Yes: all per litter
- Soft tissue examinations: Yes:
- Skeletal examinations: Yes:
- Head examinations: No data
Statistics:
Multiple sample χ ^ 2- test was performed on findings that showed one level of positive grade among sexual cycle, mating rate, conception rate, childbirth rate and nursing rate, and histopathological examination results, and in case of significant 2 Sample c 2 - Assay was performed. In addition, Fisher's direct probability test method was used when these tests failed. For other observations and findings showing positive grade of 2 or more out of the results of histopathological examination, after equality dispersal test of Bartlett, analyzed by one way analysis of variance or Kruskal-Wallis method, significant , The comparison group and the administration group were compared by Dunnett's test method or Mann-Whitney U-test method. For the test with the control group, the significance level was set at 5%.
Indices:
No data available
Historical control data:
No data available
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Stuy 1.In males, hair removal and crusts were observed in one patient in the 250 mg / kg group from 37 days to the autopsy date, but were accidental, not seen in other groups.
In females, no abnormality was observed in any of pre-pregnancy administration period, pregnancy period and nursing period.
Study 2.No effect on behaviour and appearance of treated female rats were observed as compared to control.
Dermal irritation (if dermal study):
not specified
Mortality:
no mortality observed
Description (incidence):
Study 2.No effect on survival of treated rats were observed as compared to control
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Study 1.No significant difference was observed between treated group and control group in any of period of male administration, female pre-pregnancy administration period, pregnancy period and nursing period.
Study 2.No effect on body weight gain of treated female rats were observed as compared to control.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Study 1.In males, low food intake was observed on day 46 of administration in the 500 mg / kg group, but it was a transient change with no dose correlation.
In females, there was no significant difference in treated administration group in each pregnancy administration period, pregnancy period and nursing period compared with the control group in each group.
Food efficiency:
effects observed, treatment-related
Description (incidence and severity):
Study 1.In males, low food intake was observed on day 46 of administration in the 500 mg / kg group, but it was a transient change with no dose correlation.
In females, there was no significant difference in treated administration group in each pregnancy administration period, pregnancy period and nursing period compared with the control group in each group.
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:
no effects observed
Description (incidence and severity):
Study 1.There was no significant difference in any of the test items compared with the control group in each group of male
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
Study 1.In males, no significant difference was observed in each treated group and control group.
In the females, the high weight of the body weight ratio of the lung was in the 250 mg / kg group and the high value of the weight and weight ratio of the spleen was observed in the 500 mg / kg group, but in the 1000 mg /kg group compared with the control group There was no significant difference.
Gross pathological findings:
not specified
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
Study 1.Histopathological examination revealed eosinophilic corpuscle of the proximal tubular epithelium in the kidney in males in 1 to 3 cases in the 250, 500 and 1000 mg / kg groups, but in comparison with the control group No significant difference was observed and it was not considered to be an effect of test material administration no dose-dependent increase was observed in the collection of lung foam cells and the number of occurrences of lymphocyte infiltration of the prostate, and the other findings were also changed only in one case, which is not related to test material administration. Histological examination of females showed only one case in the 1000 mg / kg group, but peripallal fatty liver in the liver, fatty degeneration of the proximal tubular epithelium in the kidney and atrophy of the thymus were observed in the kidney
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Number of abortions:
not specified
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
no effects observed
Early or late resorptions:
no effects observed
Dead fetuses:
no effects observed
Changes in pregnancy duration:
not specified
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): no effects observed
Changes in number of pregnant:
no effects observed
Other effects:
effects observed, treatment-related
Description (incidence and severity):
Study 1.There was no effect of test material administration on maternal necropsy, pregnancy period, number of corpus luteums, implantation number, implantation rate, birth rate, delivery rate, number of births and number of surviving children and birth rate at birth confirmation. However, in one case of the 1000 mg / kg group, among 9 males and 8 females, 1 male and 1 female died on 2 nursing, 2 male on 3 nursing and 1 male female on nursery 4.
Dose descriptor:
NOAEL
Effect level:
250 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
body weight and weight gain
changes in number of pregnant
clinical signs
early or late resorptions
food consumption and compound intake
gross pathology
histopathology: non-neoplastic
mortality
organ weights and organ / body weight ratios
pre and post implantation loss
total litter losses by resorption
urinalysis
Remarks on result:
other: No toxic effects obaserved
Abnormalities:
not specified
Localisation:
not specified
Fetal body weight changes:
effects observed, treatment-related
Description (incidence and severity):
Study 1.Compared with the control group, the low value of body weight of 4 days of nursing in males of the 500 mg / kg group, the 4 days of nursing of body weight gain, the weight gain and the body weight gain rate were low in females as compared with the control group. Even in the 1000 mg / kg group, low values of weight of body
weight of 4 days of nursing were observed in male, 4 days of body weight in nursing and low value of body weight gain were observed in females.
Study 2.No effects on body weight of fetous were observed as compared to control.
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): not specified
Reduction in number of live offspring:
no effects observed
Description (incidence and severity):
Study 2.No effect on viability of fetous were observed as compared to control.
Changes in sex ratio:
not specified
Changes in litter size and weights:
not specified
Changes in postnatal survival:
no effects observed
External malformations:
effects observed, treatment-related
Description (incidence and severity):
Study 1.Deficits in the tail tips of the control group observed in general condition observation and one in each of the 1000 mg / kg group, and abdominal or cervical trauma in 1 case of each sex of 1000 mg / kg group were observed , No other abnormalities were found in any of the dead cases and autopsy of newborn sacrificed on the 4th day of nursing in any animals.
Skeletal malformations:
no effects observed
Visceral malformations:
no effects observed
Other effects:
not specified
Dose descriptor:
NOAEL
Effect level:
250 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
not specified
Basis for effect level:
fetal/pup body weight changes
changes in postnatal survival
external malformations
Remarks on result:
other: No developmental toxic effects were observed
Abnormalities:
not specified
Localisation:
other: not specified
Developmental effects observed:
not specified
Treatment related:
not specified
Relation to maternal toxicity:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified

Influence of test material on reproductive performances of rats in combined repeat dose and reproductive/ developmental toxicity screening test

 

Item

0mg/kg

250mg/kg

500mg/kg

1000mg/kg

No of animals examined

12

12

12

12

No of pairs mated

12

12

12

12

No of pairs with successful copulation

12

12

12

12

Duration of mating (days. Mean ±SD.)

3.3± 1.1

2.9 ±1.2

3.7± 3.4

3.6 ±2.5

Copulation index(%)

100.0

100.0

100.0

100.0

No of pregnant animals

12

12

12

10

Fertility index(%)

100.0

100.0

100.0

83.3

a:(No. of pairs with successful copulation/ no. of pairs mated)x103

b:(No. of pregnant animals/ no. of pairs with successful copulation)x100

 

Influence of test material on developmental performances of rats in combined repeat dose and reproductive/ developmental toxicity screening test

Item

0 mg/kg

250mg/kg

500 mg/kg

1000 mg/kg

No of pregnant animals

12

12

12

12

No of corpora lutea

16.9± 2.4

17.5± 2.8

17.7 ±2.1(11)

18.1± 1.9

No of implantation sites

15.7 ±1.7

16.6 ±1.7

15.7± 4.2

15.5± 3.2

Implantation index‘(%)

93.3± 8.3

95.6 ±7.3

93.7 ±10.5

86.0 ±18.5

No of pups born

14.3± 1.7

15.6± 2.1

14.3 ±5.2

15.2± 3.2

Delivery index"(%)

91.2± 7.2

93.9 ±7.0

85.5± 27.3

98.2± 3.0**

Live pups born

 

 

 

 

No.

14.3± 1.7

15.4 ±2.1

15.5 ±2.6(11)

15.1 ±3.1

Live birth index‘(%)

100.0± 0.0

98.9± 2.5

99.5± 1.6(11)

99.4 ±1.8

Sex ratio (M/F)

1.21 ±0.63

1.36± 0.64

1.10 ±0.82

1.23 ±0.57

Dead pups born

 

 

 

 

No.

0.0 ±0.0

0.2± 0.4

0.1± 0.3(11)

0.1± 0.3

Gestation length(day)

22.5± 0.5

22.7± 0.5

22.7 ±0.2(11)

22.3 ±0.5

Gestation index‘(%)

100.0

100.0

91.7

100.0

Nursing index°(%)

100.0

100.0

100.0

100.0

Live pups on day 4

 

 

 

 

No

14.1 ±1.4

15.3 ±2.1

15.0 ±2.4(11)

14.1± 3.4

Viability index‘(%)

99.0 ±2.3

99.5 ±1.8

97.3 ±5.0(11)

91.1± 14.7

Body weight of pups(g)

 

 

 

 

Male Day 0

6.93 ±0.40

6.82 ±0.69

6.71 ±0.50(11)

6.41 ±0.62

Day 1

7.61 ±0.55

7.53± 0.91

7.24± 0.60(11)

6.94 ±0.94

Day 4

10.88± 0.86

10.62± 1.55

10.04 ±0.83*(11)

9.68 ±1.93*

Day 04 gain(g)

3.96 ±0.51

3.80 ±1.04

3.33 ±0.64(11)

3.27± 1.54

Body weight gain%

57.04± 5.54

55.48± 11.51

49.81± 10.29(11)

50.441 ±22.95

Female Day 0

6.58 ±0.40

6.44± 0.64

6.27 ±0.49(11)

6.09 ±0.56

Day 1

7.21± 0.47

7.06± 0.47

6.74± 0.59(11)

6.58± 0.81

Day 4

10.43 ±0.79

10.08 ±1.46

9.39 ±0.65**(11)

9.12± 1.71*

Day 0-4. gain(g)

3.85 ±0.47

3.63± 1.04

3.12± 0.46**(11)

3.03 ±1.42*

Body weight gain(%)

58.50 ±5.70

56.26 ±13.16

50.03 ±8.93*(11)

49.47± 22.16

 

Values are expressed as Mean ±S.D.

Values in parent theses are no. of animals examined.

Significantly different from 0 mg/kg group*p<0.05,**p <0.01

a:(No. of implantation sites/no. of corpora lutea)X100

b:(No of pups born/no. of implantation sites)X100

c:(No. or live pups born/ no. or pups born)X100

d:(No of females with live pups delivered/no. of pregnant females)X100

e:(No. of females nursing live pup/no. of females with normal delivery)X100

f: (No. of live pups on day 4/ no. of live pups born)X100

g:(Body weight gain/body weight on day 0)X100

Conclusions:
NOAEL was considered to be 250 mg/kg/day for F0 and F1 generation when pregnant female rats were treateed with test material orally.
Executive summary:

Data available from different studies were reviewed to determine the developmental toxicity of test material.The studies are as mentioned below:

Study 1

The reproductive and developmental toxicity of test material was performed on male and female rats. The test material suspended in 1 w / v% aqueous solution of sodium carmellose in dose concentration0, 250, 500, 1000 mg/kg bw/day and administered orally by gavage. The dose concentration were selected on the bases on dose finding study (5, 70 and 1000 mg / kg).Exposure to test material for the males, 46 days including the mating period and for the females for 14 days before mating and the mating period until mating, and further the mating trial was the period of pregnancy and 3 days of gestation. For males and females on 14th day of administration, they were allowed to live together in the same group within 1 to 1 (random combination) from the evening for only 14 days and the day on which sperm was confirmed in female vaginal plaque was taken as the 0th gestation. For all sexes, behaviours and appearance were observed by visual inspection and palpation at frequencies of once or more during the test period. For all female mated females, from the 21st day of pregnancy to the end of calving, the status of parturition, nursing behaviour, the number of total births, the number of surviving children and the number of dead children, the sex and the outer table of the infant were observed.

In males, hair removal and crusts were observed in one patient in the 250 mg / kg group from 37 days to the autopsy date, but were accidental, not seen in other groups. In females, no abnormality was observed in any of pre-pregnancy administration period, pregnancy period and nursing period.No significant difference was observed between treated group and control group in any of period of male administration, female pre-pregnancy administration period, pregnancy period and nursing period.In males, low food intake was observed on day 46 of administration in the 500 mg / kg group, but it was a transient change with no dose correlation. In females, there was no significant difference in treated administration group in each pregnancy administration period, pregnancy period and nursing period compared with the control group in each group.

In reproductive tests, male and female mating ratios, female sexual cycles and conception rates, test material administration in the necropsy, weight and histopathological examination of reproductive organs (testis, epididymis and ovaries) and endocrine organs (pituitary, adrenal) No effect was observed. On the other hand, pathological examination of the infertile reproductive organs of 2 cases observed in the 1000 mg / kg group showed no abnormality and no influence by test material administration was observed.

There was no effect of test material administration on maternal necropsy, pregnancy period, number of corpus luteums, implantation number, implantation rate, birth rate, delivery rate, number of births and number of surviving children and birth rate at birth confirmation. However, in one case of the 1000 mg / kg group, among 9 males and 8 females, 1 male and 1 female died on 2 nursing, 2 male on 3 nursing and 1 male female on nursery 4.Three cases were unknown, and the weight of surviving newborn infants was decreasing. In this mother animals, body weight loss and low food intake were also observed during the nursing period, and histopathological examination revealed peripallal fatty liver in the liver, fatty degeneration of the proximal tubular epithelium in the kidney and thymic Atrophy was observed. Therefore, although only one case appeared, the possibility that test material administration might have affected the mother's ability to nurture was considered.

In addition, in one case of 500 mg / kg group where delivery was not observed until 26th day of pregnancy, laparotomy of left and right uterus and cervix was observed at necropsy. In the same example, five implantation marks were seen by observation with the naked eye, both of which are considered to be early embryonic deaths, and the relation with test material administration was observed. survival were observed, in neonates weighing transition observed low values of body weight nursing 4 days in male and female 500 and 1000 mg / kg group, In addition, in the 500 mg / kg females, the body weight gain and the body weight gain rate were low, and even in females of the 1000 mg / kg group, the body weight gain was low. But neonatal weight during delivery was not changed in each group treated with test material. Therefore, a decrease or milk migration nursing ability of dams by test material administration was believed to have affected the newborn weighing 500 and 1000 mg / kg group, including one example of 1000 mg / kg group described above, The mechanism could not be clarifiedunder this test condition. Hence The no effect level (NOEL) for reproduction of parent animals by repetitive oral administration of test material in this study was considered to be 1,000 mg / kg / day in male, 250 mg / kg / day for females, and 250 mg / kg / day for no effect on neonatal development (NOEL).

Study 2.

In a teratogenicity study, pregnant female rats were treateed with test material in the concnetration of 0, 25, 75 and 250 mg/kg/day orally by gavage in 0.5% Methocel. No effect were observed on survival, clinical sign and body weight gain of treated female rats were observed as compared to control. Similarly, no effect on number of corpora lutea, number of implantation sites, Early and Late resorptions, number of resorptions and number of dams with resorptions of treated female rats were observed as compared to control. In addition, No effect on viability, number of offspring (normal or abnormal), sex ratio and body weight of fetous of treated female rats were observed as compared to control. No effect on gross pathology skeletal andvisceral abnormalities of fetous of treated female rats were observed as compared to control. Therefore, NOAEL was considered to be 250 mg/kg/day for F0 and F1 generation when pregnant female rats were treateed with test material orally by gavage for 10 days.

 

Based on the data available from different studies,Test material did not showed developmental toxicity at dose concentration 250mg/kg bw/day by oral route.Hence the test chemical is not likely to classify as a reproductive developmental toxicant as per the criteria mentioned in CLP regulation.

 

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
250 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
Data is Klimicsh 2 and from authoritative database
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

Data available from different studies were reviewed to determine the developmental toxicity of test material.The studies are as mentioned below:

Study 1

The reproductive and developmental toxicity of test material was performed on male and female rats. The test material suspended in 1 w / v% aqueous solution of sodium carmellose in dose concentration0, 250, 500, 1000 mg/kg bw/day and administered orally by gavage. The dose concentration were selected on the bases on dose finding study (5, 70 and 1000 mg / kg).Exposure to test material for the males, 46 days including the mating period and for the females for 14 days before mating and the mating period until mating, and further the mating trial was the period of pregnancy and 3 days of gestation. For males and females on 14th day of administration, they were allowed to live together in the same group within 1 to 1 (random combination) from the evening for only 14 days and the day on which sperm was confirmed in female vaginal plaque was taken as the 0th gestation. For all sexes, behaviours and appearance were observed by visual inspection and palpation at frequencies of once or more during the test period. For all female mated females, from the 21st day of pregnancy to the end of calving, the status of parturition, nursing behaviour, the number of total births, the number of surviving children and the number of dead children, the sex and the outer table of the infant were observed.

In males, hair removal and crusts were observed in one patient in the 250 mg / kg group from 37 days to the autopsy date, but were accidental, not seen in other groups. In females, no abnormality was observed in any of pre-pregnancy administration period, pregnancy period and nursing period.No significant difference was observed between treated group and control group in any of period of male administration, female pre-pregnancy administration period, pregnancy period and nursing period.In males, low food intake was observed on day 46 of administration in the 500 mg / kg group, but it was a transient change with no dose correlation. In females, there was no significant difference in treated administration group in each pregnancy administration period, pregnancy period and nursing period compared with the control group in each group.

In reproductive tests, male and female mating ratios, female sexual cycles and conception rates, test material administration in the necropsy, weight and histopathological examination of reproductive organs (testis, epididymis and ovaries) and endocrine organs (pituitary, adrenal) No effect was observed. On the other hand, pathological examination of the infertile reproductive organs of 2 cases observed in the 1000 mg / kg group showed no abnormality and no influence by test material administration was observed.

There was no effect of test material administration on maternal necropsy, pregnancy period, number of corpus luteums, implantation number, implantation rate, birth rate, delivery rate, number of births and number of surviving children and birth rate at birth confirmation. However, in one case of the 1000 mg / kg group, among 9 males and 8 females, 1 male and 1 female died on 2 nursing, 2 male on 3 nursing and 1 male female on nursery 4.Three cases were unknown, and the weight of surviving newborn infants was decreasing. In this mother animals, body weight loss and low food intake were also observed during the nursing period, and histopathological examination revealed peripallal fatty liver in the liver, fatty degeneration of the proximal tubular epithelium in the kidney and thymic Atrophy was observed. Therefore, although only one case appeared, the possibility that test material administration might have affected the mother's ability to nurture was considered.

In addition, in one case of 500 mg / kg group where delivery was not observed until 26th day of pregnancy, laparotomy of left and right uterus and cervix was observed at necropsy. In the same example, five implantation marks were seen by observation with the naked eye, both of which are considered to be early embryonic deaths, and the relation with test material administration was observed. survival were observed, in neonates weighing transition observed low values of body weight nursing 4 days in male and female 500 and 1000 mg / kg group, In addition, in the 500 mg / kg females, the body weight gain and the body weight gain rate were low, and even in females of the 1000 mg / kg group, the body weight gain was low. But neonatal weight during delivery was not changed in each group treated with test material. Therefore, a decrease or milk migration nursing ability of dams by test material administration was believed to have affected the newborn weighing 500 and 1000 mg / kg group, including one example of 1000 mg / kg group described above, The mechanism could not be clarifiedunder this test condition. Hence The no effect level (NOEL) for reproduction of parent animals by repetitive oral administration of test material in this study was considered to be 1,000 mg / kg / day in male, 250 mg / kg / day for females, and 250 mg / kg / day for no effect on neonatal development (NOEL).

Study 2.

In a teratogenicity study, pregnant female rats were treateed with test material in the concnetration of 0, 25, 75 and 250 mg/kg/day orally by gavage in 0.5% Methocel. No effect were observed on survival, clinical sign and body weight gain of treated female rats were observed as compared to control. Similarly, no effect on number of corpora lutea, number of implantation sites, Early and Late resorptions, number of resorptions and number of dams with resorptions of treated female rats were observed as compared to control. In addition, No effect on viability, number of offspring (normal or abnormal), sex ratio and body weight of fetous of treated female rats were observed as compared to control. No effect on gross pathology skeletal andvisceral abnormalities of fetous of treated female rats were observed as compared to control. Therefore, NOAEL was considered to be 250 mg/kg/day for F0 and F1 generation when pregnant female rats were treateed with test material orally by gavage for 10 days.

 

Based on the data available from different studies,Test material did not showed developmental toxicity at dose concentration 250mg/kg bw/day by oral route.Hence the test chemical is not likely to classify as a reproductive developmental toxicant as per the criteria mentioned in CLP regulation.

 

Justification for classification or non-classification

Thus, comparing this value with the criteria of CLP regulation test chemical is not likely to classify as reproductive and developmental toxicant.

Additional information