Registration Dossier
Registration Dossier
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EC number: 230-049-5 | CAS number: 6925-69-5
Endpoint summary
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
- 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. - 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
- 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.
Reference
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.
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.
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
- 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
- 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.
Reference
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
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
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