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 100 -180 mg/kg /day for reproductive toxicity, when male and female rats were treated with test material orally. Thus, comparing this value with the criteria of CLP regulation test material is not 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:
Weight of evidence approach based on various test chemicals
Justification for type of information:
Weight of evidence approach based on various test chemicals
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Qualifier:
according to
Guideline:
other: Weight of evidence approach based on various test chemicals
Principles of method if other than guideline:
Weight of evidence approach based on various test chemicals
GLP compliance:
not specified
Limit test:
no
Justification for study design:
not specified
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
2.Details on test animal
TEST ANIMALS- Source: IGS, Japan Chiyarusu-Lipa Inc., Atsugi breeding center.
- Age at study initiation: (P) x wks; (F1) x wks : Parent : 10 weeks old
- Weight at study initiation:
(P) Males: x-x g; Females: x-x g;
(F1) Males: x-x g; Females: x-x g :
Parent : Male 389 to 452 g Female 219 to 266 g
- Fasting period before study: No data available
- Housing: Animals were housed one per by sex in a metal net cage, during the mating period two per sex per cage will mother were housed individually in plastic Ekonkeji you turn on the (white flakes Japan Chiyarusu River Co., Ltd.)
- Diet (e.g. ad libitum): solid feed (NMF, Oriental Yeast Co., Ltd.), ad libitum.
- Water (e.g. ad libitum): Drinking water (NMF, Oriental Yeast Co., Ltd.), ad libitum.
- Acclimation period: 19 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23 ± 3 ℃
- Humidity (%): 50 ± 20 %
- Air changes (per hr): 10-15 times per hour ventilation.
- Photoperiod (hrs dark / hrs light): 12 hours dark / 12 hours light
Route of administration:
other: 2.oral: gavage; 3.oral: feed
Type of inhalation exposure (if applicable):
not specified
Vehicle:
water
Remarks:
Purified water
Details on exposure:
2. Details on exposure
PREPARATION OF DOSING SOLUTIONS:
Test material dissolved in Water for injection
DIET PREPARATION
- Rate of preparation of diet (frequency):No data available
- Mixing appropriate amounts with (Type of food)
- Storage temperature of food: No data available

3. Details on exposure
PREPARATION OF DOSING SOLUTIONS: Undiluted test material given by oral route in food
DIET PREPARATION
- Rate of preparation of diet (frequency):No data available
- Mixing appropriate amounts with (Type of food )
- Storage temperature of food: No data available
VEHICLE
- Justification for use and choice of vehicle (if other than water): none
- Concentration in vehicle: 0, 20, 50 and 100 mg /kg bw/day
- Amount of vehicle (if gavage): No data available

- Lot/batch no. (if required): No data available
- Purity: No data available
Details on mating procedure:
2.- M/F ratio per cage: 1:1 ratio
- Length of cohabitation: Overnight
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancyConfirmed the sperm in the pupil plaque or during Hitomi plug formation considered as day 0 of pregnancy.
- After … days of unsuccessful pairing replacement of first male by another male with proven fertility. No data available
- Further mating 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
3.- M/F ratio per cage: No data available
- Length of cohabitation: 21 days
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy No data available
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility.
- Further matings after two unsuccessful attempts: [no / yes (explain)] No data available
- After successful mating each pregnant female was caged (how): No data available
- Any other deviations from standard protocol: No data available
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
2.Male : 42days
Female : 41 - 53 days (from 14 days before mating to day 4 of lactation)
3.starting 10 weeks before mating (F0) or at weaning (F1), throughout mating, gestation and lactation periods until scheduled necropsy
Frequency of treatment:
2,3. Daily
Details on study schedule:
3.F0 rats were mated twice to produce F1a and F1b litters. Twenty-eight parents/sex of the F2 generation were selected from the F1b litters. They were mated twice to produce F2a and F2b litters. Mating was allowed for up to 21 days. Necropsy was performed on 10 pups/sex/dose from the F1a and F1b generation
Remarks:
Study 2.Doses / Concentrations: 0, 60, 180 or 600 mg/kg/dayBasis:actual ingested
Remarks:
Study 3.0, 20, 50 and 100 mg /kg bw/day
No. of animals per sex per dose:
2.Total:116
Test group
0 mg/kg bw/day:12 male and 12 female
60mg/kg bw/day:12 male and 12 female
180mg/kg bw/day:12 male and 12 female
600mg/kg bw/day:12 male and 12 female
Recovery group
0 mg/kg bw/day:5 male and 5 female
0 mg/kg bw/day:5male and 5 female
3.Total:448
For F0 generation
0 mg/kg bw/day:28 male and 28 female
20mg/kg bw/day:28 male and 28 female
50mg/kg bw/day:28 male and 28 female
100 mg/kg bw/day:28 male and 28 female
For F1 generation
0 mg/kg bw/day:28 male and 28 female
20mg/kg bw/day:28 male and 28 female
50mg/kg bw/day:28 male and 28 female
100 mg/kg bw/day:28 male and 28 female
Control animals:
yes, concurrent vehicle
Details on study design:
2. Dose selection rationale:
14-day repeated oral dose toxicity study using rats for the test chemical (dose setting test)" was carried out earlier dose (dose: 100,300,1000 mg / kg). 1 rat died on the 14th administration in male 1000 mg / kg dose group, in a general state of deaths loose stools (reddish brown), body weight and food consumption was incresed in 300 mg/kg, liver weight was observed high, low value of the ALP, high total protein, expansion of the cecum at autopsy in male and female were observed at 1000 mg/kg daoe gruop. Therefore, the dose level of 0, 60, 180 and 600 mg/kg/bw/day was chosen in the current study.
- Rationale for animal assignment (if not random): Animals were randomized by body weight.
- Rationale for selecting satellite groups: 5 female as satellite group.
- Post-exposure recovery period in satellite groups: No data available
- Section schedule rationale (if not random): No data available
Positive control:
No data available
Parental animals: Observations and examinations:
2. Observations and examinations performed & frequency
CAGE SIDE OBSERVATIONS: Yes
Attitude, fistula attack, abnormal behaviour were observed.

- Time schedule: Once, once a week (but 1,7,14 and 20 days of gestation, mating confirmation female animals nursing the 4th calving animals) were observed.

- Cage side observations checked in table [No.?] were included: On 1,7,14 and 20 days of gestation, mating confirmation female animals nursing the 4th calving animals were observed.

DETAILED CLINICAL OBSERVATIONS: Yes
Pupil diameter, abnormal breathing, the state of the fur-skin, secretions of the eyes, nose, exophthalmos,flair closed state, the visible mucous membranes, autonomic function and urination were observed.

- Time schedule: 3 times daily

BODY WEIGHT: Yes
Throughout the recovery period and the period of administration.

- Time schedule for examinations:
At a frequency of twice a week, check mating female animals were weighed on the 4th and lactation, as well as 0 and 20 days 0, 4, 7, 11, 14, 17 pregnancy.

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

- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data available

- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data available

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No data available

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data available

- Time schedule for examinations: No data available

OPHTHALMOSCOPIC EXAMINATION: Yes
Pupil diameter and secretions of the eyes were observed.

- Time schedule for examinations: Daily

- Dose groups that were examined: 0, 60, 180 and 600 mg/kg body weight/day

HAEMATOLOGY: Yes

- Time schedule for collection of blood:
Before autopsy.

- Anaesthetic used for blood collection: Yes

- Animals fasted: Yes
On day 43, overnight (16-20 hours) fasted.
- How many animals: each 5 per sex per group

- Parameters checked in table [No.?] were examined. : RBC, Hb, Ht, MCV, MCH, MCHC, Reticulocyte, Platelet, PT, APTT, Fibrinogen, WBC and differential leucocytes were examined.

CLINICAL CHEMISTRY: Yes / No / No data
- Time schedule for collection of blood:
- Animals fasted: Yes / No / No data
- How many animals:
- Parameters checked in table [No.?] were examined.

URINALYSIS: Yes
Urinalysis of male rat were observed on administration of Tetrabutylammonium bromide and the last week of recovery, urine collected from 5 animals in each group and observed the color, pH, protein, ketone bodies, glucose, occult blood, bilirubin, urobilinogen and urine volume.

- Time schedule for collection of urine: Daily 20 hour.

- Metabolism cages used for collection of urine: No data available

- Animals fasted: Yes
For 4 hours

- Parameters checked in table [No.?] were examined.:
Color, pH, protein, ketone bodies, glucose, occult blood, bilirubin, urobilinogen and urine volume were checked.

NEUROBEHAVIOURAL EXAMINATION: No data available

- Time schedule for examinations: No data available

- Dose groups that were examined: n No data available

- Battery of functions tested: sensory activity / grip strength / motor activity / other:
Grip strength: Grip strength measurement of hindlimb and forelimb were tested.

Spontaneous momentum: Spontaneous movement sensor was counting the momentum of 60 minutes and 10 minute intervals.

OTHER: Auditory response, approaching response, contact reaction, pain reaction, anti-morphism air righting reflex, landing leg width was observed.
3. Parental animals observation and examinations
CAGE SIDE OBSERVATIONS: yes

DETAILED CLINICAL OBSERVATIONS: Yes

Time schedule:

BODY WEIGHT: Yes
Time schedule for examinations:
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes Food consumption was determined weekly.

Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes / No / No data: No data available


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

OTHER:
Oestrous cyclicity (parental animals):
Any irregularities in the estrous cycle were investigated.
Sperm parameters (parental animals):
No data available
Litter observations:
2. STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: [yes/no]
- If yes, maximum of [...] pups/litter ([...]/sex/litter as nearly as possible); excess pups were killed and discarded.

PARAMETERS EXAMINED
The following parameters were examined in [F1 / F2 / F3] offspring:
[number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain, physical or behavioural abnormalities, anogenital distance (AGD), pup weight on the day of AGD, presence of nipples/areolae in male pups, other. Particular attention should be paid to the external reproductive genitals which should be examined for signs of altered development; gross evaluation of external genitalia]
: Litter were observed for survival rate, external abnormalities, body weight, number of live pups and sex ratio

GROSS EXAMINATION OF DEAD PUPS:
[no / yes, for external and internal abnormalities; possible cause of death was/was not determined for pups born or found dead]

ASSESSMENT OF DEVELOPMENTAL NEUROTOXICITY:

ASSESSMENT OF DEVELOPMENTAL IMMUNOTOXICITY:
Postmortem examinations (parental animals):
2. Sacrifice and pathology
GROSS PATHOLOGY: Yes
Change attributed in intestinal tracts of male and female of 180 and 600 mg/kg/bw/day group and in liver of male and female of 600 mg/kg/bw/day group.

HISTOPATHOLOGY: Yes
Diffuse hyperplasis in mucosa in cecum,which thought to be related to the macroscopic dilation in lunina of cecum, in the rectum, the debris in crypt were observed in female of 600 mg/kg/bw/day group.

In recovery group of 600 mg/kg/bw/day cell infertilation in mucosa were observed cecum,colon and rectum. In the liver , hypertropy of preiobular hepatocyte were observed.
3. SACRIFICE
- Male animals: All surviving animals [describe when, e.g. as soon as possible after the last litters in each generation were produced.]
- Maternal animals: All surviving animals [describe when, e.g. after the last litter of each generation was weaned.]

GROSS NECROPSY: yes
- Gross necropsy consisted of [external and internal examinations including the cervical, thoracic, and abdominal viscera.]

HISTOPATHOLOGY / ORGAN WEIGHTS: yes
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.
Postmortem examinations (offspring):
2. GROSS NECROPSY
- Gross necropsy consisted of [external and internal examinations including the cervical, thoracic, and abdominal viscera.]: External observation and necropsy.
3. SACRIFICE
- The F1 offspring not selected as parental animals and all F2 offspring were sacrificed at [#?] days of age.
- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows:

GROSS NECROPSY:yes
- Gross necropsy consisted of [external and internal examinations including the cervical, thoracic, and abdominal viscera.]

HISTOPATHOLOGY / ORGAN WEIGTHS
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.
Statistics:
Statistical analysis was carried out by using x 2 test with continuity correction of Yeates, expected frequency was observed cell of 5 or less, by the direct probability calculation method of Fisher test (significance level for the 4th). Dunnett type test (mean rank test method) (significance level 0,06 ) performed for homogeneity of variance of each group and Bartlett method firstly test was applied. (significance level 0.01, both sides)
Reproductive indices:
Copulation index, Fertility index, Gestation index, Delivery index and Live birth index.
Offspring viability indices:
Viability (survival rate) on Day 4 after birth was recorded.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
2.In 1 male (600 mg/kg group, dead animal) Fracture of incisor, soft stool and decrease in the amount of feces was observed. While in 1 female Staining of lower abdominal fur, staggering gait ((600 mg/kg group ,dead animal).
3.No treatment-related clinical signs were observed
Dermal irritation (if dermal study):
not specified
Mortality:
mortality observed, treatment-related
Description (incidence):
2.Deaths occurred in 1 male and 1 pregnant female in the 600 mg/kg group.
3.There were no treatment related deaths during the study. One male at each dosing level was killed due to trauma or caging accident or sacrificed moribund.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
2.In the 600 mg/kg group, males showed suppressed body weight gain and females showed tendencies toward high values in body weight and food consumption during the administration period. A high value in food consumption was also observed in females in the 180 mg/kg group. Males in the 600 mg/kg group also showed a low value in body weight during the recovery period. However, since the body weight gain in this group during the recovery period was comparable to that of the control group, they were thought to have recovery.
3.Body weight gain was reduced in males and females at 100 mg/kg/day food during the premating period.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
3.Food consumption was reduced at 100 mg/kg /day during the premating period in males and females.
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:
effects observed, treatment-related
Description (incidence and severity):
2.In blood chemistry examination, males in the 600 mg/kg group showed a high value in AST and a low value in blood urea nitrogen and females showed high values in AST and LDH and low values in blood urea nitrogen and creatinine. The females in the 600 mg/kg group showed a high value in calcium as well. These changes were not observed after the end of the recovery period showing reversibility of the changes.
Urinalysis findings:
effects observed, treatment-related
Description (incidence and severity):
2.some males in the 600 mg/kg group showed red urine sporadically and light brown urine was observed at urinalysis, but red urine was a transient change.
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
3.No treatment-related abnormalities were observed, including histopathological examinations
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Reproductive function: oestrous cycle:
effects observed, treatment-related
Description (incidence and severity):
3.In the 600 mg/kg group, the number of females that showed abnormalities in estrous cycle tended to be high,
Reproductive function: sperm measures:
not specified
Reproductive performance:
effects observed, treatment-related
Description (incidence and severity):
2.In parent animals in the 60 and 180 mg/kg groups, there were no test article-related changes in the number of days until copulation, copulation index, insemination index or fertility index. There
were no test article-related changes in the delivery index, length of gestation period, number of
corporalutea, number of implantation sites, implantation index, number of live born pups or sex ratio, and there were no test article-related changes in parturition condition or lactation observations.

In the 600 mg/kg group, the number of females that showed abnormalities in estrous cycle tended to be high, the number of days until copulation tended to be low, copulation index, insemination index or fertility index tended to be low values, a high value in stillborn index, and low values in viability of pups (day 0 and day 4 of lactation)
3.There were no changes detected between parental animals of the treated and control groups in mating indices and pregnancy rates. No treatment-related changes were detected in litter size, live birth index, viability index, lactation index and sex ratio of the F0 offspring
Results of examination: Parental Clinical signs and Mortality:Deaths occurred in 1 male and 1 pregnant female when treated with 600 mg/kg/day. Clinical signs: Before death, the male showed fractures of incisors, soft stool and a decreased in the amount of feces produced. The female showed staining of lower abdominal fur and staggering gait before death.
Body weight and food consumption:
Body weight: When treated with 600 mg/kg/day, the males showed suppressed body weight gain while females showed a tendency toward high values in body weight during the administration period. In comparison to control, males in the 600 mg/kg/day group showed a low value in body weight during the recovery period.
Food consumption: Females showed a tendency toward high values in food consumption during the administration period. A high value in food consumption was also observed in females treated with 180 mg/kg/day.
Test substance intake: No data available
Reproductive function-
Estrous cycle: Treatment with 600 mg/kg/day in females resulted in abnormalities in estrous cycle, which tended to be high. In addition, the number of days until copulations, copulation index, insemination index or fertility index tended to be low.
Reproductive function-
Reproductive performanceThe 600 mg/kg/day group showed high value in the stillborn index.
Organ weights: In males treated with 600 mg/kg/day, the absolute organ weight was decreased for thymus, heart, kidneys and epididymis. In females treated with 600 mg/kg/day, the absolute organ weight was decreased for brain and thymus, while the organ weight for thyroids, heart, liver, spleen and kidneys increased.
Gross pathology: Change attributed in intestinal tracts of male and female of 180 or 600 mg/kg/day groups and in liver of male and female of the 600 mg/kg/day group. Dilatation in lumina of the cecum was also observed.
Histopathology: Diffuse hyperplasia in mucosa in the cecum was observed. In rectum, cell debris in crypt was observed in females. In the recovery group, these changes were no longer observed. However in the recovery group, cell infiltration in mucosa was observed in cecum, colon and rectum. In the liver, hypertrophy of perilobular hepatocytes was observed. other findingsHematologyMales treated with 600 mg/kg/day showed a slight decrease in platelets. Similar effects were observed during the recovery period.An increased level of fibrinogen was also seen in the same animals, i.e. the 600 mg/kg/day-treated males and males from the recovery period. Males in recovery period also showed decreased levels of lymphocyte and eosinophils while the level of neutrophils and monocytes were increased.Females treated with 600 mg/kg/day showed a slight increase in RBC and a slight decrease in platelets. These effects were not observed during the recovery period.In addition, a decreased level of fibrinogen was also seen in the same animals, i.e. the 600 mg/kg/day-treated females. Females in recovery period showed decreased levels of lymphocytes and increased levels of monocytes.
Clinical chemistry: When treated with 600 mg/kg/day, males showed a high value in AST and a low value in blood urea nitrogen, while females showed high values in AST and LDH and low values in blood urea nitrogen and creatinine. During treatment high values of calcium was also observed in these females, however, the high levels of calcium was not detected after the recovery period. UrinanalysisIn male rat treated with 600 mg/kg/day showed red urine sporadically and light brown urine were observed.
Dose descriptor:
NOAEL
Effect level:
> 100 - <= 180 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Effects on body weight, survival rate, organ weight, hematology, urinalysis and histopathology.
Remarks on result:
other: No toxic effetcs were observed
Dose descriptor:
LOAEL
Effect level:
600 mg/kg bw/day
Based on:
test mat.
Sex:
female
Basis for effect level:
reproductive function (oestrous cycle)
reproductive performance
Remarks on result:
other: effects observed on reproductive performance
Critical effects observed:
not specified
System:
other: not specified
Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality / viability:
mortality observed, treatment-related
Description (incidence and severity):
2.Low value of survival were observed in pups with parent from the 600 mg/kg/day group.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
2.In the 600 mg/kg group, low values in the body weight of males and females at the time of birth and on day 4 after birth, and male and female pups showed a low value in the body weight gain during the lactation period.
3.F1 pup weight gains were initially similar to those in the control group, but were significantly decreased on day 21 and 28 post-partum at 100 mg/kg /day (82-87% and 83-86% of control for F1a males and females, and 76-85% and 78-83% of control for F1b males and females, resp.).
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
no effects observed
Description (incidence and severity):
2.In the examination of newborn pups, there were no test article-related changes in external observation or necropsy on day 4 after birth
3.F1a and F1b pups showed no treatment-related macroscopic findings.
Histopathological findings:
not specified
Other effects:
not specified
Behaviour (functional findings):
not specified
Developmental immunotoxicity:
not specified
Results of examination of Offspring:
Clinical signs: No data available
Body weight and food consumption-
Body weight: Male and female pups showed a low value in the body weight gain during the lactation period.
Food consumption No data available
Test substance intake: No data available
Reproductive function- e
Estrous cycle: No data available
Reproductive function-
Sperm measures: Reproductive performance: Low value of survival were observed in pups with parent from the 600 mg/kg/day group.
Organ weights: No data available
Gross pathology: No data available
Histopathology: No data available
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
> 50 - <= 180 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
gross pathology
other: Effects on body weight and survival rate.
Remarks on result:
other: No toxic effects were observed
Dose descriptor:
LOAEL
Generation:
F1
Effect level:
600 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
viability
Remarks on result:
other: Low value of survival were observed in pups with parent from the 600 mg/kg/day group.
Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality / viability:
not specified
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
3. F2a pup weight gains were decreased at 100 mg/kg /day on day 21 and 28 post-partum (85-89% and 83-90% of control for males and female resp.). F2b pup weight gains were decreased at 100 mg/kg/day on day 21 and 28 post-partum (85-89% and 84-86% of control).
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
no effects observed
Description (incidence and severity):
3. F2a and F2b pups showed no treatment-related macroscopic finding
Histopathological findings:
not specified
Other effects:
not specified
Developmental immunotoxicity:
not specified
Dose descriptor:
NOAEL
Generation:
F2
Effect level:
50 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
not specified
Basis for effect level:
viability
body weight and weight gain
gross pathology
Remarks on result:
other: No developmental toxicity was observed
Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Reproductive effects observed:
not specified
Treatment related:
not specified
Conclusions:
Based on the available results and applying the weight of evidence approach, the NOAEL for the test chemical lies between 40 -180 mg/kg/day in both the Parental and F1 generation when rats were exposed to the test chemical.
Executive summary:

Various studies have been reviewed to determine the reproductive toxicity potential of the test chemical. These include in vivo experimental studies carried out on rats for the test chemical. The results are mentioned below:

Combined Repeated Dose Toxicity Study with the Reproduction/ Developmental Toxicity Screening Test was performed for the test chemical to evaluate its toxicity potential.The male and female Crl:CD (SD)rats were treated withtest chemicalin dose concentrations of 0, 60, 180, 600 mg/kg/day, Recovery 0, 600 mg/kg/day (R600) in Water for injection by oral gavage route for 42 days in male and 53 days (from 14 days before mating to day 4 of lactation) in female.12 animals /sex / dose group were used in test group and 5 animals /sex / dose in the recovery group. All the animals were observed for clinical signs, toxicity and body weight and food consumption. The male sacrificed on day 43 while female sacrificed on day 5 of lactation. Organ weight, gross pathological examination, histopathlogical examination, haematological examination was performed. Reproductive parameter like to the copulation index, gestation length, delivery conditions, nursing conditions, fertility index, and number of corpora lutea, implantation rate, or gestation index were noted.Deaths occurred in 1 male and 1 pregnant female in the 600 mg/kg group.In 1 male (600 mg/kg group ,dead animal)Fracture of incisor, soft stool and decrease in the amount of feces was observed. While in 1 female Staining of lower abdominal fur, staggering gait ((600 mg/kg group ,dead animal).In the 600 mg/kg group, males showed suppressed body weight gain and females showed tendencies toward high values in body weight and food consumption during the administration period. A high value in food consumption was also observed in females in the 180 mg/kg group. Males in the 600 mg/kg group also showed a low value in body weight during the recovery period. However, since the body weight gain in this group during the recovery period was comparable to that of the control group, they were thought to have recovery.In blood chemistry examination, males in the 600 mg/kg group showed a high value in AST and a low value in blood urea nitrogen and females showed high values in AST and LDH and low values in blood urea nitrogen and creatinine. The females in the 600 mg/kg group showed a high value in calcium as well. These changes were not observed after the end of the recovery period showing reversibility of the changes.Some males in the 600 mg/kg group showed red urine sporadically and lightbrown urine was observed at urinalysis, but it was considered as a transient change.Macroscopically, dilatation in lumina of the cecum was observed. Histopathologically, diffuse hyperplasia in mucosa in the cecum, which was thought to be related to the macroscopic dilatation in lumina of the cecum, was observed. In the rectum, cell debris in crypt was observed in females. In the recovery group, the above changes were no longer observed and thus recovery from these changes was indicated In the recovery group, cell infiltration in mucosa was observed in the cecum, colon and rectum. In the liver, hypertrophy of perilobular hepatocytes was observed. In parent animals in the 60 and 180 mg/kg groups, there were no test chemical-related changes in the number of days until copulation, copulation index, insemination index or fertility index. There were no test chemical-related changes in the delivery index, length of gestation period, number ofcorpora lutea, number of implantation sites, implantation index, number of live born pups or sex ratio, and there were no test chemical-related changes in parturition condition or lactation observations. In the 600 mg/kg group, the number of females that showed abnormalities in estrouscycle tended to be high, the number of days until copulation tended to be low, copulation index, insemination index or fertility index tended to be low values, a high value in stillborn index, and low values in viability of pups (day 0 and day 4 of lactation) .In the examination of new born pups, there were no test article-related changes in external observation or necropsy on day 4 after birth.In the 600 mg/kg group, low values in the body weight of males and females at the time of birth and on day 4 after birth, and male and female pups showed a low value in the body weight gain during the lactation period. Hence, the No Observed Adverse Effect Level (NOAEL) was considered to be 180mg/kg/day, on the bases of reproductive parameter and developmental effects.When male and femalerats were treated withtest chemical orally.

This result is supported by a two generation reproduction study of test chemical was performed on male and female Sprague Dawley CD® rats in accordance with OECD guideline 416 (1983). Although oestrus cycle, sperm parameters and sexual maturation of pups was not investigated, no reproductive effects were observed indicating indirectly no effect on these parameters. The study is considered acceptable. The test chemical mixed with food in dose concentrations 0, 20, 50 and 100 mg /kg bw/day (0, 300, 750 and 1500 mg/kg food ).28 male and 28 female placed in each dose group. Diet starting 10 weeks before mating (F0) or at weaning (F1), throughout mating, gestation and lactation periods until scheduled necropsy. F0 rats were mated twice to produce F1a and F1b litters. Twenty-eight parents/sex of the F2 generation were selected from the F1b litters. They were mated twice to produce F2a and F2b litters. Mating was allowed for up to 21 days. Necropsy was performed on 10 pups/sex/dose from the F1a and F1b generation. Clinical signs, mortality, body weight and Food consumption were noted in each generation. 

 No treatment-related clinical signs were observed. There were no treatment related deaths during the study. One male at each dosing level was killed due to trauma or caging accident or sacrificed moribund. Body weight gain was reduced in males and females at 100 mg/kg/day food during the premating period. Food consumption was reduced at 100 mg/kg /day during the premating period in males and females. At necropsy no treatment-related abnormalities were observed, including histopathological examinations there were no changes detected between parental animals of the treated and control groups in mating indices and pregnancy rates. No treatment-related changes were detected in litter size, live birth index, viability index, lactation index and sex ratio of the F0 offspring.

 F1 pup weight gains were initially similar to those in the control group, but were significantly decreased on day 21 and 28 post-partum at 100 mg/kg /day (82-87% and 83-86% of control for F1a males and females, and 76-85% and 78-83% of control for F1b males and females, resp.). F1a and F1b pups showed no treatment-related macroscopic findings. F1 parental animals showed no treatment-related mortality or clinical signs. Body weight gain was only incidentally reduced for F1 parents. Body weights of males and females at 100 mg/kg /day were already significantly reduced at the beginning of mating due to the reduced body weight gain during their lactation period. Body weights of females were still decreased during gestation and were equal to controls at the end of lactation due 10 increased body weight gains relative to controls during gestation and lactation (F28 and F2b). Food consumption was significantly reduced at 100mg/kg /day in F1 males until week 7 and F1 females during the whole pre-mating period. No changes were noted between parental animals of the treated and control groups in mating indices, pregnancy rates and male fertility. At necropsy no treatment-related abnormalities were observed, including histopathological examination. No treatment-related changes were observed in litter size, live birth index, viability index,l actation index or sex ratio of the F, offspring (F2pups). F2apup weight gains were decreased at 100 mg/kg /day on day 21 and 28 post-partum (85-89% and 83-90% of control for males and female resp.). F2bpup weight gains were decreased at 100 mg/kg/day on day 21 and 28post-partum (85-89%and 84-86% of control).F2a andF2b pups showed no treatment-related macroscopic finding. Hence No Observed Adverse Effect Level (NOAEL) for F0 generation was considered to be 100mg/kg/day, As No treatment related changes were noted in mating behaviour, conception and gestation and the NOAEL for developmental toxicity was considered to be 50 mg/kg /day, based on decreased body weight. When female Sprague Dawley rats were treated with test material orally.

Based on the available results and applying the weight of evidence approach, the NOAEL for the test chemical lies between 40 -180 mg/kg/day in both the Parental and F1 generation when rats were exposed to the test chemical.

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
100 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

Various studies have been reviewed to determine the reproductive toxicity potential of the test chemical. These include in vivo experimental studies carried out on rats for the test chemical. The results are mentioned below:

Combined Repeated Dose Toxicity Study with the Reproduction/ Developmental Toxicity Screening Test was performed for the test chemical to evaluate its toxicity potential.The male and female Crl:CD (SD)rats were treated withtest chemicalin dose concentrations of 0, 60, 180, 600 mg/kg/day, Recovery 0, 600 mg/kg/day (R600) in Water for injection by oral gavage route for 42 days in male and 53 days (from 14 days before mating to day 4 of lactation) in female.12 animals /sex / dose group were used in test group and 5 animals /sex / dose in the recovery group. All the animals were observed for clinical signs, toxicity and body weight and food consumption. The male sacrificed on day 43 while female sacrificed on day 5 of lactation. Organ weight, gross pathological examination, histopathlogical examination, haematological examination was performed. Reproductive parameter like to the copulation index, gestation length, delivery conditions, nursing conditions, fertility index, and number of corpora lutea, implantation rate, or gestation index were noted.Deaths occurred in 1 male and 1 pregnant female in the 600 mg/kg group.In 1 male (600 mg/kg group ,dead animal)Fracture of incisor, soft stool and decrease in the amount of feces was observed. While in 1 female Staining of lower abdominal fur, staggering gait ((600 mg/kg group ,dead animal).In the 600 mg/kg group, males showed suppressed body weight gain and females showed tendencies toward high values in body weight and food consumption during the administration period. A high value in food consumption was also observed in females in the 180 mg/kg group. Males in the 600 mg/kg group also showed a low value in body weight during the recovery period. However, since the body weight gain in this group during the recovery period was comparable to that of the control group, they were thought to have recovery.In blood chemistry examination, males in the 600 mg/kg group showed a high value in AST and a low value in blood urea nitrogen and females showed high values in AST and LDH and low values in blood urea nitrogen and creatinine. The females in the 600 mg/kg group showed a high value in calcium as well. These changes were not observed after the end of the recovery period showing reversibility of the changes.Some males in the 600 mg/kg group showed red urine sporadically and lightbrown urine was observed at urinalysis, but it was considered as a transient change.Macroscopically, dilatation in lumina of the cecum was observed. Histopathologically, diffuse hyperplasia in mucosa in the cecum, which was thought to be related to the macroscopic dilatation in lumina of the cecum, was observed. In the rectum, cell debris in crypt was observed in females. In the recovery group, the above changes were no longer observed and thus recovery from these changes was indicated In the recovery group, cell infiltration in mucosa was observed in the cecum, colon and rectum. In the liver, hypertrophy of perilobular hepatocytes was observed. In parent animals in the 60 and 180 mg/kg groups, there were no test chemical-related changes in the number of days until copulation, copulation index, insemination index or fertility index. There were no test chemical-related changes in the delivery index, length of gestation period, number ofcorpora lutea, number of implantation sites, implantation index, number of live born pups or sex ratio, and there were no test chemical-related changes in parturition condition or lactation observations. In the 600 mg/kg group, the number of females that showed abnormalities in estrouscycle tended to be high, the number of days until copulation tended to be low, copulation index, insemination index or fertility index tended to be low values, a high value in stillborn index, and low values in viability of pups (day 0 and day 4 of lactation) .In the examination of new born pups, there were no test article-related changes in external observation or necropsy on day 4 after birth.In the 600 mg/kg group, low values in the body weight of males and females at the time of birth and on day 4 after birth, and male and female pups showed a low value in the body weight gain during the lactation period. Hence, the No Observed Adverse Effect Level (NOAEL) was considered to be 180mg/kg/day, on the bases of reproductive parameter and developmental effects.When male and femalerats were treated withtest chemical orally.

This result is supported by a two generation reproduction study of test chemical was performed on male and female Sprague Dawley CD® rats in accordance with OECD guideline 416 (1983). Although oestrus cycle, sperm parameters and sexual maturation of pups was not investigated, no reproductive effects were observed indicating indirectly no effect on these parameters. The study is considered acceptable. The test chemical mixed with food in dose concentrations 0, 20, 50 and 100 mg /kg bw/day (0, 300, 750 and 1500 mg/kg food ).28 male and 28 female placed in each dose group. Diet starting 10 weeks before mating (F0) or at weaning (F1), throughout mating, gestation and lactation periods until scheduled necropsy. F0 rats were mated twice to produce F1a and F1b litters. Twenty-eight parents/sex of the F2 generation were selected from the F1b litters. They were mated twice to produce F2a and F2b litters. Mating was allowed for up to 21 days. Necropsy was performed on 10 pups/sex/dose from the F1a and F1b generation. Clinical signs, mortality, body weight and Food consumption were noted in each generation. 

 No treatment-related clinical signs were observed. There were no treatment related deaths during the study. One male at each dosing level was killed due to trauma or caging accident or sacrificed moribund. Body weight gain was reduced in males and females at 100 mg/kg/day food during the premating period. Food consumption was reduced at 100 mg/kg /day during the premating period in males and females. At necropsy no treatment-related abnormalities were observed, including histopathological examinations there were no changes detected between parental animals of the treated and control groups in mating indices and pregnancy rates. No treatment-related changes were detected in litter size, live birth index, viability index, lactation index and sex ratio of the F0 offspring.

 F1 pup weight gains were initially similar to those in the control group, but were significantly decreased on day 21 and 28 post-partum at 100 mg/kg /day (82-87% and 83-86% of control for F1a males and females, and 76-85% and 78-83% of control for F1b males and females, resp.). F1a and F1b pups showed no treatment-related macroscopic findings. F1 parental animals showed no treatment-related mortality or clinical signs. Body weight gain was only incidentally reduced for F1 parents. Body weights of males and females at 100 mg/kg /day were already significantly reduced at the beginning of mating due to the reduced body weight gain during their lactation period. Body weights of females were still decreased during gestation and were equal to controls at the end of lactation due 10 increased body weight gains relative to controls during gestation and lactation (F28 and F2b). Food consumption was significantly reduced at 100mg/kg /day in F1 males until week 7 and F1 females during the whole pre-mating period. No changes were noted between parental animals of the treated and control groups in mating indices, pregnancy rates and male fertility. At necropsy no treatment-related abnormalities were observed, including histopathological examination. No treatment-related changes were observed in litter size, live birth index, viability index,l actation index or sex ratio of the F, offspring (F2pups). F2apup weight gains were decreased at 100 mg/kg /day on day 21 and 28 post-partum (85-89% and 83-90% of control for males and female resp.). F2bpup weight gains were decreased at 100 mg/kg/day on day 21 and 28post-partum (85-89%and 84-86% of control).F2a andF2b pups showed no treatment-related macroscopic finding. Hence No Observed Adverse Effect Level (NOAEL) for F0 generation was considered to be 100mg/kg/day, As No treatment related changes were noted in mating behaviour, conception and gestation and the NOAEL for developmental toxicity was considered to be 50 mg/kg /day, based on decreased body weight. When female Sprague Dawley rats were treated with test material orally.

Based on the available results and applying the weight of evidence approach, the NOAEL for the test chemical lies between 40 -180 mg/kg/day in both the Parental and F1 generation when rats were exposed to the test chemical.

Effects on developmental toxicity

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available

Justification for classification or non-classification

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