Registration Dossier

Administrative data

Key value for chemical safety assessment

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 600mg /kg bw/day.When rats or rabbits 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:
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:
read-across source
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Qualifier:
equivalent or similar to
Guideline:
other: As mentioned below
Principles of method if other than guideline:
WoE report is based on reproductive toxicity studies on rats or rabbits
GLP compliance:
not specified
Limit test:
no
Justification for study design:
No data available
Species:
other: 1& 2.rats 3.rabbits
Strain:
other: 1.Wistar:Han (TM): RCCHan(TM): WIST 2.Sprague-Dawley 3.New Zealand White
Remarks:
Han (TM): RCCHan(TM): WIST 2.Sprague-Dawley
Details on species / strain selection:
No data available
Sex:
male/female
Details on test animals and environmental conditions:
3.Details on test animals and env. conditions
TEST ANIMALS
- Source: Hazelton Research Products, I n c . , Denver. PA
- Age at study initiation: (P) approximately 6 mos
- Weight at study initiation: Females: 2490-4440g
- Fasting period before study: No data available
- Housing: Animals were individually hosed in stainless steel cage with mesh flooring (Hoeltge. In c .Cincinnat,, OH).
- Use of restrainers for preventing ingestion (if dermal):no
- Diet (e.g. ad libitum): Purina certified Rabbit chow@ (#5322) pellets , ad libitum
- Water (e.g. ad libitum): deionized/filtered water ad libitum
- Acclimation period:14 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C):18.33-18.66°C
- Humidity (%):58%-64%
- Air changes (per hr): No data available
- Photoperiod (hrs dark / hrs light): lights were on from 07.00h to 19.00h.

Route of administration:
oral: gavage
Vehicle:
other: 1.Arachis oil BP 2.corn oil
Details on exposure:
1.Details on exposure
PREPARATION OF DOSING SOLUTIONS: test material soluble in Arachis oil BP.
DIET PREPARATION
- Rate of preparation of diet (frequency):Fresh diets were made and distributed weekly
- 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): Arachis oil BP
- Concentration in vehicle: 0, 30, 300 and 600mg/kg bw/day
- Amount of vehicle (if gavage):
- Lot/batch no. (if required): No data available
- Purity: No data available
2.Details on exposure
PREPARATION OF DOSING SOLUTIONS:
Test material dissolved in corn oil
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): Test material dissolved in corn oil
- Concentration in vehicle: 0,30,100,300mg/kg bw/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 materials dissolved in corn oil. The corn oil used i n the dose formulations contained less than 2 mill equivalents of peroxide/kg corn oil.
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): Test material dissolved in corn oil and stable throughout the study.
- Concentration in vehicle: 0, 50,200,400mg/kg bw/day
- Amount of vehicle (if gavage): 2ml/kg bw
- Lot/batch no. (if required): No data available
- Purity: No data available
Details on mating procedure:
1.- M/F ratio per cage:1:1
- Length of cohabitation:
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy
- 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:
3.A artificially inseminated female animals were used
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
1.11 week ( (including a two week pre-pairing phase, pairing, gestation and early lactation for females)
2.During premating, mating, gestation, and lactation.
3.13 days ( gestational days 6-19)
Frequency of treatment:
Daily
Details on study schedule:
No data available
Remarks:
Study1.
0, 30,300,600 mg/kg bw/day
Study 2.
0, 30,100,300mg/kg /day
Study 3.
0, 50, 200, 400 mg/kg bw/day
No. of animals per sex per dose:
Study 1
Total:80
0 mg/kg bw/day: 10 male and 10 female
30mg/kg bw/day: 10 male and 10 female
300mg/kg bw/day: 10 male and 10 female
600mg/kg bw/day: 10 male and 10 female
Study 2.
Total:100
0 mg/kg bw/day:25
30mg/kg bw/day:25
100mg/kg bw/day:25
300 mg/kg bw/day:25
Study 3.
Total:104
0 mg/kg bw/day:26
50mg/kg bw/day:26
200mg/kg bw/day:26
400 mg/kg bw/day:26
Control animals:
yes, concurrent vehicle
Details on study design:
No data available
Positive control:
No data available
Parental animals: Observations and examinations:
1.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 for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes


WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): yes
2.Parental animals observation and examinations
CAGE SIDE OBSERVATIONS: yes

DETAILED CLINICAL OBSERVATIONS: Yes

Time schedule: daily


BODY WEIGHT: Yes
Time schedule for examinations: daily
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:
3.Parental animals observation and examinations
CAGE SIDE OBSERVATIONS: yes

DETAILED CLINICAL OBSERVATIONS:

Time schedule: Animals were observed daily before (gd 0-5). During (gd 6-19). and after (gd 20-30) dosing
For clinical signs of toxicity .

BODY WEIGHT: Yes
Time schedule for examinations: animals were weighed on the mornings of gd 0, 6 through 19,25 and 30
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes Food was weighed on gd 0,3,6,9,12,15,18,19,22,25,28 and 30.
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:


Oestrous cyclicity (parental animals):
No data available
Sperm parameters (parental animals):
1.Parameters examined in [all/P/F1/F2] male parental generations:
testis weight, and spermatogenetic cycle were observed
2.Parameters examined in [all/P/F1/F2] male parental generations:
sperm assessments for motility and concentration
Litter observations:
1.Offspring litter size, sex ratio and viability were observed
2.STANDARDISATION OF LITTERS:After weaning, 25 F1 generation pups/sex/dosage group were randomly selected for evaluation until sexual maturity.
3.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. No data available

PARAMETERS EXAMINED
The following parameters were examined in [F1] offspring:
number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies were observed
GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities; possible cause of death was/was not determined for pups born or found dead

ASSESSMENT OF DEVELOPMENTAL NEUROTOXICITY:No data available

ASSESSMENT OF DEVELOPMENTAL IMMUNOTOXICITY:No data available


PARAMETERS EXAMINED
The following parameters were examined in [F1] offspring:
viability, body weights, sexual maturation, anogenital distance (days 1 and 22 postpartum), nipple eruption (day 12 postpartum) were observed

GROSS EXAMINATION OF DEAD PUPS:yes
gross necropsy observations were recorded.

ASSESSMENT OF DEVELOPMENTAL NEUROTOXICITY:No data available

ASSESSMENT OF DEVELOPMENTAL IMMUNOTOXICITY:No data available
Postmortem examinations (parental animals):
1.Postmortem examinations (Parent Animal)
SACRIFICE :
Male animals: Adult males were terminated on Day 52 of the study following the completion of the second pairing at 600 mg/kg bw/day
Maternal animals: yes
Females were terminated on day 5 post partum. Any female which did not produce a pregnancy (unless allocated to a further mating phase) was terminated on or after Day 25 post coitum.

GROSS NECROPSY: yes

HISTOPATHOLOGY / ORGAN WEIGHTS
histopathological evaluation of reproductive tissues was performed. Additional male organ weight and detailed histopathological examination of the testes were performed to more fully assess male fertility.
2.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 was performed

HISTOPATHOLOGY / ORGAN WEIGHTS: yes
3.SACRIFICE
- Male animals: All surviving animals [describe when, e.g. as soon as possible after the last litters in each generation were produced.]No data available
- Maternal animals: All surviving animals [describe when, e.g. after the last litter of each generation was weaned.]The animals were killed with an injection of sodium pentobarbital (Barber Veterinary Supply. Fayetteville NC) in the marginal ear vein on gd 30


GROSS NECROPSY: yes
At gross necropsy Liver and intact uterus was weighed and corpora lutea were counted. Uterine contents were examined to determine the number of implantation sites, resorptions, dead foetuses and live foetuses. Uteri which had no visible implantation sites was stained with ammonium sulphide (10%) to detect very early resorptions

HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.

HISTOPATHOLOGY / ORGAN WEIGTHS
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.
Postmortem examinations (offspring):
1.Postmortem examinations (offspring)
SACRIFICE
Offspring were terminated on day 5 post partum.
GROSS NECROPSY
Yes
HISTOPATHOLOGY / ORGAN WEIGTHS
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively
2.SACRIFICE
- The F1 offspring were sacrificed at 22 days of age.
- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows:

GROSS NECROPSY: yes
- Gross necropsy was performed

HISTOPATHOLOGY / ORGAN WEIGTHS: No data available
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.
3.SACRIFICE:live foetuses were dissected from the uterus and also anesthetized with sodium pentobarbital.

- 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:
General Linear Models (GLM) procedures were applied for the analyses of variance (ANOVA) of maternal and fetal parameters. Prior to GLM analysis ,an arcsine-square root transformation was performed on a litter- derived percentage data (Snedecor and Cochran. 1967) and Bartletts test for homogeneity of variance was performed on all data to be analysed by ANOVA (Winer. 196'2). GLM analysis determined the significance of dose-response relationships and the significance of dose effects. Replicate effects and dose x replicate interactions. When ANOVA revealed a significant (p<0.05) dose effect. Williams' and Dunnett's Multiple Comparison Tests (Williams, 1971: 1972: Dunnett. 1955: 1964) compared CAM-exposed to control groups. One-tailed tests were used for all pairwise comparisons except maternal body and organ weights. Maternal food consumption. fetal body weight and percent males per litter. Non-significant (p>0.05) dose x replicate effects on selected fetal parametric measures were considered justification for pooling data across replicates for nonparametric analysis on related measures. When a significant (p<0.05#) dose x replicate' interaction occurred, the data for that endpoint and f0.r any related nominal scale data were analysed separately for dose effects within each replicate in the study. As well as for all replicates combined. Nominal scale measures were analysed by a x2 test for independence and by a test 'f0.r linear trend on proportions. When x2 test showed significant group differences. a one tailed Fisher's exact probability test was used for pairwise comparisons of CAM and control groups.
Reproductive indices:
No data available
Offspring viability indices:
No data available
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
1.Transient post-dosing salivation was observed from Day 7 at 600 mg/kg bw/day and Day 13 at 300 mg/kg bw/day. This sign was observed regularly throughout the treatment period with all animals being affected at both dosages, although the incidence of this finding was greatest at the high dosage. Transient post dosing salivation was also observed at 30 mg/kg bw/day for two females on Day 2 and one male on Day 50 of this study.
2.Increased incidences of excess salivation occurred in P generation male and female rats at 100 and/or 300 mg/kg/d throughout the dosage period but these clinical signs were not considered as adverse effects of test material administration.
Dermal irritation (if dermal study):
not specified
Mortality:
no mortality observed
Description (incidence):
1.There were no unscheduled deaths in the study.
2.No incedence of mortality was noted
3.There were no dose related maternal deaths during the study, but two does were removed from the 400 mg/kg/day dose group because of dosing error.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
1.At 600 mg/kg bw/day body weight gain of males was statistically significantly lower than control during the first week of treatment. Subsequent body weight gains were considered to reflect normal biological variation, but overall gain at termination remained lower than control.
Bodyweight gain of males at 30 and 300 mg/kg bw/day and for females at all dosages, throughout the pre-pairing, gestation and lactation phases of the study, were considered to have been unaffected by treatment.
2. no effects on body weight was observed.
3.Maternal body weight before, during and after treatment period in the treated animals was also comparable to control weights at all gestational ages.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
1.Food consumption for both sexes was considered to have been unaffected by treatment throughout the study, and including gestation and lactation phases for females, at 30, 300 and 600 mg/kg bw/day.
2.no effects were observed on food consumption in male and female rats.
3.Food consumption in all dose -treated groups was similar to control throughout the study.
Food efficiency:
effects observed, treatment-related
Description (incidence and severity):
Food efficiency: At 600 mg/kg bw/day food conversation efficiency for males was lower than control during week 1; subsequent food utilisation was similar to control. Food conversation efficiency of males at 30 and 300 mg/kg bw/day and for females at all dosages, throughout the pre-pairing, gestation and lactation phases of the study, were considered to have been unaffected by the treatment.
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
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
1.Male reproductive organ weights were unaffected by treatment at 30, 300 and 600 mg/kg bw/day and did not indicate any effect on fertility
2.Organ weight remain unaffected.
3.Gravid uterine and absolute and relative maternal liver weights were similar to vehicle control values.
Gross pathological findings:
no effects observed
Description (incidence and severity):
2. No relevant gross pathological findings were recorded.
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
Histopathological examinations did not indicate any effect of treatment at 600 mg/kg bw/day and these examinations, including detailed assessment of the spermatogenetic cycle for the testes did not indicate any effect on male fertility
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Reproductive function: oestrous cycle:
not specified
Reproductive function: sperm measures:
no effects observed
Description (incidence and severity):
2.no effects were observed for sperm assessments for motility and concentration.
Reproductive performance:
no effects observed
Description (incidence and severity):
1.Mating: Pre-coital interval and mating evidence at the times of conception did not indicate any adverse effect of treatment on mating performance at 30, 300 and 600 mg/kg bw/day. Fertility: At 600 mg/kg bw/day, only seven females delivered a litter following the initial pairing but subsequent re-mating and additional assessment of male organ weight and detailed testicular histopathology did not indicate any treatment related effect on fertility for either sex. There was also no effect of treatment on fertility at 30 and 300 mg/kg bw/day. Gestation lengths: Gestation length was considered to be unaffected by treatment at 30, 300 and 600 mg/kg bw/day. Litter responses: Offspring litter size, sex ratio and viability: There was no effect of maternal treatment on corpora lutea and implantations counts, pre- and postimplantation
loss, number of offspring born, sex ratio or subsequent survival to Day 4 of age at 30, 300 and 600 mg/kg bw/day.
2.No effects were observed on mating and fertility and litter observations. Also natural delivery and ovarian follicle counts were unaffected in treated group animals.
Dose descriptor:
NOAEL
Effect level:
> 300 - <= 600 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
food efficiency
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
reproductive function (sperm measures)
reproductive performance
Remarks on result:
other: No toxic effects were 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.Assessment of surface righting ability on Day 1; offspring clinical signs did not indicate any effect of maternal treatment
2.no effects on clinical signs were noted
Dermal irritation (if dermal study):
not specified
Mortality / viability:
no mortality observed
Description (incidence and severity):
2.No effects on viability was observed.
3.Live litter size were unaffected .
Body weight and weight changes:
no effects observed
Description (incidence and severity):
1.At 600 mg/kg bw/day initial offspring body weight was similar to control but weight gain to Day 4 was statistically significantly lower than control. Mean offspring body weights, litter weight and weight gains to Day 4 of age were unaffected by maternal treatment at 30 and 300 mg/kg bw/day.
3.average fetal body weight were unaffected
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
Description (incidence and severity):
2.no effects observed on sexual maturation
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
no effects observed
Description (incidence and severity):
1.Macroscopic necropsy findings did not indicate any effect of treatment at 30, 300 and 600 mg/kg bw/day.
2. No relevant gross pathological findings were recorded.
3.No external ,skeletal and visceral malformations were observed .
Histopathological findings:
not specified
Other effects:
no effects observed
Description (incidence and severity):
1.No dose related effects were observed on anogenital distance (days 1 and 22 postpartum), nipple eruption (day 12 postpartum),
3.No incidence of anatomical variation or defects were observed.
Behaviour (functional findings):
not specified
Developmental immunotoxicity:
not specified
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
> 300 - <= 600 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
not specified
Basis for effect level:
clinical signs
body weight and weight gain
gross pathology
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
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:
No Observed Adverse Effect Level (NOAEL) for reproductive toxicity was considered to 600mg/kg/day, When male and female rats orrabbits were treated with test material orally.
Executive summary:

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

Study 1

The reproductive and developmental toxicity study of test material was performed on male and femaleWistar Han (TM): RCCHan(TM): WIST strain rats. The test material was dissolved in Arachis oil BP and administered in dose concentration 0, 30,300,600mg/kg bw /day via oral gavage for up to eleven weeks (including a two week pre-pairing phase, pairing, gestation and early lactation for females).Each treatment group contain 10 male and 10 female rats while a control group of ten males and ten females was dosed with vehicle alone (Arachis oil BP). Clinical signs, bodyweight change, dietary intake and water consumption were monitored during the study. Pairing of animals within each dose group was undertaken on a one male: one female basis within each treatment group on Day 5 of the study, with females subsequently being allowed to litter and rear their offspring to Day 5 of lactation. An additional pairing for high dose females that failed to achieve pregnancy was performed to fully assess mating performance and fertility. During the lactation phase, daily clinical observations were performed on all surviving offspring, together with litter size and offspring weights and assessment of surface righting reflex. Adult males were terminated on Day 52 of the study following the completion of the second pairing at 600 mg/kg bw/day. Females and offspring were terminated on day 5 post partum. Any female which did not produce a pregnancy (unless allocated to a further mating phase) was terminated on or after Day 25 post coitum. All animals were subjected to a gross necropsy examination and histopathological evaluation of reproductive tissues was performed. Additional male organ weight and detailed histopathological examination of the testes were performed to more fully assess male fertility.

 

There were no unscheduled deaths in the study.Transient post-dosing salivation was observed from Day 7 at 600 mg/kg bw/day and Day 13 at 300 mg/kg bw/day. This sign was observed regularly throughout the treatment period with all animals being affected at both dosages, although the incidence of thisfinding was greatest at the high dosage. Transient post dosing salivation was also observed at 30 mg/kg bw/day for two females on Day 2 and one male on Day 50 of this study.At 600 mg/kg bw/day body weight gain of males was statistically significantly lower than control during thefirst week of treatment. Subsequent body weight gains were considered to reflect normal biological variation, but overall gain at termination remained lower than control.

Bodyweight gain of males at 30 and 300 mg/kg bw/day and for females at all dosages, throughout the pre-pairing, gestation and lactation phases of the study, were considered to have been unaffected by treatment. Food consumption for both sexes was considered to have been unaffected by treatment throughout the study, and including gestation and lactation phases for females, at 30, 300 and 600 mg/kg bw/day. Food efficiency: At 600 mg/kg bw/day food conversation efficiency for males was lower than control during week 1; subsequent food utilisation was similar to control. Food conversation efficiency of males at 30 and 300 mg/kg bw/day and for females at all dosages, throughout the pre-pairing, gestation and lactation phases of the study, were considered to have been unaffected by the treatment. Male reproductive organ weights were unaffected by treatment at 30, 300 and 600 mg/kg bw/day and did not indicate any effect on fertility. Histopathological examinations did not indicate any effect of treatment at 600 mg/kg bw/day and these examinations, including detailed assessment of the spermatogenetic cycle for the testes did not indicate any effect on male fertility. Pre-coital interval and mating evidence at the times of conception did not indicate any adverse effect of treatment on mating performance at 30, 300 and 600 mg/kg bw/day. Fertility: At 600 mg/kg bw/day, only seven females delivered a litter following the initial pairing but subsequent re-mating and additional assessment of male organ weight and detailed testicular histopathology did not indicate any treatment related effect on fertility for either sex. There was also no effect of treatment on fertility at 30 and 300 mg/kg bw/day. Gestation length was considered to be unaffected by treatment at 30, 300 and 600 mg/kg bw/day. Litter responses: Offspring litter size, sex ratio and viability: There was no effect of maternal treatment on corpora lutea and implantations counts, pre- and postimplantation loss, number offspring born, sex ratio or subsequent survival to Day 4 of age at 30, 300 and 600 mg/kg bw/day. Assessment of surface righting ability on Day 1; offspring clinical signs and necropsyfindings and did not indicate any effect of maternal treatment. Assessment of surface righting ability on Day 1; offspring clinical signs and necropsyfindings and did not indicate any effect of maternal treatment.Macroscopic necropsy findings did not indicate any effect of treatment at 30, 300 and 600 mg/kgbw/day. HenceNo Observed Adverse Effect Level (NOAEL) for reproductive toxicity was considered to 600mg/kg/day,When male and femalerats were treated with test material orally for 11weeks.

 Study 2.

Reproductive toxicity of test material was investigated in a 1-generation reproduction study according toOECD Guideline 415in which 25 Crl: CD (Sprague-Dawley) rats/sex/group were gavaged with dosages of 0 (corn oil vehicle), 30, 100, or 300 mg/kg/d during premating, mating, gestation, and lactation. After weaning, 25 F1 generation pups/sex/dosage group were randomly selected for evaluation until sexual maturity. In P generation males and females viability, clinical signs, body weights, feed consumption, mating and fertility, organ weights, gross and microscopic observations, sperm assessments (motility and concentration), natural delivery , litter and ovarian follicle counts were observations,. In F1 generation pups, viability, body weights, sexual maturation, anogenital distance (days 1 and 22 postpartum), nipple eruption (day 12 postpartum), and gross necropsy observations were recorded.

 

Increased incidences of excess salivation occurred in P generation male and female rats at 100 and/or 300 mg/kg/d throughout the dosage period, and low incidences of urine-stained abdominal fur were seen in females at 300 mg/kg/d during the gestation period. These clinical signs were not considered as adverse effects of test material administration. No effects were observed for viability, clinical signs, body weights, feed consumption, mating and fertility, organ weights, gross and microscopic observations, sperm assessments (motility and concentration), natural delivery , litter and ovarian follicle counts in male and female animals in parpental generation. Also in F1 generation pups, viability, body weights, sexual maturation, anogenital distance (days 1 and 22 postpartum), nipple eruption (day 12 postpartum), and gross necropsy observations were unaffected . Thus, the NOAEL for reproductive toxicity in the P generation rats and the NOAEL for viability and growth of the F1 generation offspring is considered to be ≥300 mg/kg/d.When male and femaleSprague Dawley rats were treated with test material orally.

Study 3.

The reproductive and developmental toxicity study of test materialwas performed in new Zealand white rabbits.104 rabbits were divided as 26/ dose group. The test material dissolved in corn oil and administered in dose volume 2ml /kg in concentration0, 50, 200, 400 mg/kg bw/day by oral gavage routeon gestational day 6 through 19.The designated dose level were based upon

Preliminary dose rang – finding study. A artificially inseminated female animals were used and the day of insemination was designated as Gestational day (gd) 0. Animals were observed daily before (gd 0-5),during (gd 6-19) and after (gd 20-30) dosing for clinical signs of toxicity.Animalswere weighed on the mornings of gd 0, 6 through 19,25and 30.Food was weighed on gd 0,3,6,9,12,15,18,19,22,25,28 and 30. At necropsy,The animals were killed with an injection of sodium pentobarbital(Barber Veterinary Supply. Fayetteville NC)in the marginal ear vein on gd 30. Liver and intact uterus was weighed and corpora lutea were counted. Uterine contents were examined to determine the number of implantation sites, resorptions, dead foetuses and live foetuses. Uteri which had no visible implantation sites was stained with ammonium sulphide (10%) to detect very early resorptions.live foetuses were dissected from the uterus and also anesthetized with sodium pentobarbital.The foetuses were weighed, sexed and subjected to external, soft tissue or skeletal examinations.Half of the foetuses were decapitated prior to dissection, the heads were fixed in bouins solution and then examined by free hand sectioning technique . All fetal carcasses (50% without heads) were stained with alcian blue/ alizarin red S and examined for skeletal malformations

 There were no dose related maternal deaths during the study, but two animals were removed from the 400 mg/kg/day dose group because of dosing error. Food consumption in all dose -treated groups was similar to control throughout the study. Maternal body weight before, during and after treatment period in the treated animals was also comparable to control weights at all gestational ages. Maternal weight gain during the dosing period tended to decrease with increasing dose. Maternal weight gain relative to control animals was reduced 13%, 5% and59%in the 50, 200 and 400 mg/kg/day dose groups respectively. Gravid uterine and absolute and relative maternal liver weights were similar to vehicle control values.

Results of the uterine examination revealed that test material was not developmental toxic effects even at the highest dose. No effect was seen for the percent resorption per litter. The percent late fetal deaths per litter , the percent non live implants per litter or the percent adversely affected implants per litter. The proportion of litters with one or more resorption , late fetal deaths, nonlive implants or adversely affected implants were not affected by dose exposure. Live litter size and average fetal body weight were also unaffected .No external , skeletal and visceral malformations were observed . No incidence of anatomical variation or defects were observed. HenceNo Observed Adverse Effect Level (NOAEL) for maternal and foetal toxicity was considered to be 400 mg/kg/day.When femalenew Zealand white rabbits were treated with test material orally.

 

Based on the data available from different studies test chemical did not showedreproductive toxicityat dose concentration 600mg/kg bw/day.When rats or rabbits were treated with test material orally.Hence the test chemical is not likely to classify as a reproductive toxicant as per the criteria mentioned in CLP regulation.

 

 

 

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
600 mg/kg bw/day
Study duration:
chronic
Species:
other: Rats or rabbits
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

Reproductive toxicity study

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

Study 1

The reproductive and developmental toxicity study of test material was performed on male and femaleWistar Han (TM): RCCHan(TM): WIST strain rats. The test material was dissolved in Arachis oil BP and administered in dose concentration 0, 30,300,600mg/kg bw /day via oral gavage for up to eleven weeks (including a two week pre-pairing phase, pairing, gestation and early lactation for females).Each treatment group contain 10 male and 10 female rats while a control group of ten males and ten females was dosed with vehicle alone (Arachis oil BP). Clinical signs, bodyweight change, dietary intake and water consumption were monitored during the study. Pairing of animals within each dose group was undertaken on a one male: one female basis within each treatment group on Day 5 of the study, with females subsequently being allowed to litter and rear their offspring to Day 5 of lactation. An additional pairing for high dose females that failed to achieve pregnancy was performed to fully assess mating performance and fertility. During the lactation phase, daily clinical observations were performed on all surviving offspring, together with litter size and offspring weights and assessment of surface righting reflex. Adult males were terminated on Day 52 of the study following the completion of the second pairing at 600 mg/kg bw/day. Females and offspring were terminated on day 5 post partum. Any female which did not produce a pregnancy (unless allocated to a further mating phase) was terminated on or after Day 25 post coitum. All animals were subjected to a gross necropsy examination and histopathological evaluation of reproductive tissues was performed. Additional male organ weight and detailed histopathological examination of the testes were performed to more fully assess male fertility.

 

There were no unscheduled deaths in the study.Transient post-dosing salivation was observed from Day 7 at 600 mg/kg bw/day and Day 13 at 300 mg/kg bw/day. This sign was observed regularly throughout the treatment period with all animals being affected at both dosages, although the incidence of thisfinding was greatest at the high dosage. Transient post dosing salivation was also observed at 30 mg/kg bw/day for two females on Day 2 and one male on Day 50 of this study.At 600 mg/kg bw/day body weight gain of males was statistically significantly lower than control during thefirst week of treatment. Subsequent body weight gains were considered to reflect normal biological variation, but overall gain at termination remained lower than control.

Bodyweight gain of males at 30 and 300 mg/kg bw/day and for females at all dosages, throughout the pre-pairing, gestation and lactation phases of the study, were considered to have been unaffected by treatment. Food consumption for both sexes was considered to have been unaffected by treatment throughout the study, and including gestation and lactation phases for females, at 30, 300 and 600 mg/kg bw/day. Food efficiency: At 600 mg/kg bw/day food conversation efficiency for males was lower than control during week 1; subsequent food utilisation was similar to control. Food conversation efficiency of males at 30 and 300 mg/kg bw/day and for females at all dosages, throughout the pre-pairing, gestation and lactation phases of the study, were considered to have been unaffected by the treatment. Male reproductive organ weights were unaffected by treatment at 30, 300 and 600 mg/kg bw/day and did not indicate any effect on fertility. Histopathological examinations did not indicate any effect of treatment at 600 mg/kg bw/day and these examinations, including detailed assessment of the spermatogenetic cycle for the testes did not indicate any effect on male fertility. Pre-coital interval and mating evidence at the times of conception did not indicate any adverse effect of treatment on mating performance at 30, 300 and 600 mg/kg bw/day. Fertility: At 600 mg/kg bw/day, only seven females delivered a litter following the initial pairing but subsequent re-mating and additional assessment of male organ weight and detailed testicular histopathology did not indicate any treatment related effect on fertility for either sex. There was also no effect of treatment on fertility at 30 and 300 mg/kg bw/day. Gestation length was considered to be unaffected by treatment at 30, 300 and 600 mg/kg bw/day. Litter responses: Offspring litter size, sex ratio and viability: There was no effect of maternal treatment on corpora lutea and implantations counts, pre- and postimplantation loss, number offspring born, sex ratio or subsequent survival to Day 4 of age at 30, 300 and 600 mg/kg bw/day. Assessment of surface righting ability on Day 1; offspring clinical signs and necropsyfindings and did not indicate any effect of maternal treatment. Assessment of surface righting ability on Day 1; offspring clinical signs and necropsyfindings and did not indicate any effect of maternal treatment.Macroscopic necropsy findings did not indicate any effect of treatment at 30, 300 and 600 mg/kgbw/day. HenceNo Observed Adverse Effect Level (NOAEL) for reproductive toxicity was considered to 600mg/kg/day,When male and femalerats were treated with test material orally for 11weeks.

 

Study 2.

Reproductive toxicity of test material was investigated in a 1-generation reproduction study according toOECD Guideline 415in which 25 Crl: CD (Sprague-Dawley) rats/sex/group were gavaged with dosages of 0 (corn oil vehicle), 30, 100, or 300 mg/kg/d during premating, mating, gestation, and lactation. After weaning, 25 F1 generation pups/sex/dosage group were randomly selected for evaluation until sexual maturity. In P generation males and females viability, clinical signs, body weights, feed consumption, mating and fertility, organ weights, gross and microscopic observations, sperm assessments (motility and concentration), natural delivery , litter and ovarian follicle counts were observations,. In F1 generation pups, viability, body weights, sexual maturation, anogenital distance (days 1 and 22 postpartum), nipple eruption (day 12 postpartum), and gross necropsy observations were recorded.

Increased incidences of excess salivation occurred in P generation male and female rats at 100 and/or 300 mg/kg/d throughout the dosage period, and low incidences of urine-stained abdominal fur were seen in females at 300 mg/kg/d during the gestation period. These clinical signs were not considered as adverse effects of test material administration. No effects were observed for viability, clinical signs, body weights, feed consumption, mating and fertility, organ weights, gross and microscopic observations, sperm assessments (motility and concentration), natural delivery , litter and ovarian follicle counts in male and female animals in parpental generation. Also in F1 generation pups, viability, body weights, sexual maturation, anogenital distance (days 1 and 22 postpartum), nipple eruption (day 12 postpartum), and gross necropsy observations were unaffected . Thus, the NOAEL for reproductive toxicity in the P generation rats and the NOAEL for viability and growth of the F1 generation offspring is considered to be ≥300 mg/kg/d.When male and femaleSprague Dawley rats were treated with test materialorally.

Study 3.

The reproductive and developmental toxicity study of test materialwas performed in new Zealand white rabbits.104 rabbits were divided as 26/ dose group. The test material dissolved in corn oil and administered in dose volume 2ml /kg in concentration0, 50, 200, 400 mg/kg bw/day by oral gavage routeon gestational day 6 through 19.The designated dose level were based upon

Preliminary dose rang – finding study. A artificially inseminated female animals were used and the day of insemination was designated as Gestational day (gd) 0. Animals were observed daily before (gd 0-5),during (gd 6-19) and after (gd 20-30) dosing for clinical signs of toxicity.Animalswere weighed on the mornings of gd 0, 6 through 19,25and 30.Food was weighed on gd 0,3,6,9,12,15,18,19,22,25,28 and 30. At necropsy,The animals were killed with an injection of sodium pentobarbital(Barber Veterinary Supply. Fayetteville NC)in the marginal ear vein on gd 30. Liver and intact uterus was weighed and corpora lutea were counted. Uterine contents were examined to determine the number of implantation sites, resorptions, dead foetuses and live foetuses. Uteri which had no visible implantation sites was stained with ammonium sulphide (10%) to detect very early resorptions.live foetuses were dissected from the uterus and also anesthetized with sodium pentobarbital.The foetuses were weighed, sexed and subjected to external, soft tissue or skeletal examinations.Half of the foetuses were decapitated prior to dissection, the heads were fixed in bouins solution and then examined by free hand sectioning technique . All fetal carcasses (50% without heads) were stained with alcian blue/ alizarin red S and examined for skeletal malformations

 There were no dose related maternal deaths during the study, but two animals were removed from the 400 mg/kg/day dose group because of dosing error. Food consumption in all dose -treated groups was similar to control throughout the study. Maternal body weight before, during and after treatment period in the treated animals was also comparable to control weights at all gestational ages. Maternal weight gain during the dosing period tended to decrease with increasing dose. Maternal weight gain relative to control animals was reduced 13%, 5% and59%in the 50, 200 and 400 mg/kg/day dose groups respectively. Gravid uterine and absolute and relative maternal liver weights were similar to vehicle control values.

Results of the uterine examination revealed that test material was not developmental toxic effects even at the highest dose. No effect was seen for the percent resorption per litter. The percent late fetal deaths per litter , the percent non live implants per litter or the percent adversely affected implants per litter. The proportion of litters with one or more resorption , late fetal deaths, nonlive implants or adversely affected implants were not affected by dose exposure. Live litter size and average fetal body weight were also unaffected .No external , skeletal and visceral malformations were observed . No incidence of anatomical variation or defects were observed. HenceNo Observed Adverse Effect Level (NOAEL) for maternal and foetal toxicity was considered to be 400 mg/kg/day.When femalenew Zealand white rabbits were treated with test material orally.

 

Based on the data available from different studies test chemical did not showedreproductive toxicityat dose concentration 600mg/kg bw/day.When rats or rabbits were treated with test material orally.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 600 mg /kg bw/day .When rats or rabbits were treated with test chemical orally. Thus, comparing this value with the criteria of CLP regulation test chemical is not likely to classify as reproductive and developmental toxicant.

Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Remarks:
Experimental data from various test chemicals
Justification for type of information:
Weight of evidence approach based on the available information from various test chemicals.
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Qualifier:
equivalent or similar to
Guideline:
other: As mentioned below
Principles of method if other than guideline:
WoE report is based on developmental toxicity studies on rats or rabbits

GLP compliance:
not specified
Limit test:
no
Species:
other: 1 &2.rats 3.rabbits
Strain:
other: 1.&2.Wistar 3.New Zealand White
Details on test animals and environmental conditions:
3.Details on test animals and env. conditions
TEST ANIMALS
- Source: Hazelton Research Products, I n c . , Denver. PA
- Age at study initiation: (P) approximately 6 mos
- Weight at study initiation: Females: 2490-4440g
- Fasting period before study: No data available
- Housing: Animals were individually hosed in stainless steel cage with mesh flooring (Hoeltge. In c .Cincinnat,, OH).
- Use of restrainers for preventing ingestion (if dermal):no
- Diet (e.g. ad libitum): Purina certified Rabbit chow@ (#5322) pellets , ad libitum
- Water (e.g. ad libitum): deionized/filtered water ad libitum
- Acclimation period:14 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C):18.33-18.66°C
- Humidity (%):58%-64%
- Air changes (per hr): No data available
- Photoperiod (hrs dark / hrs light): lights were on from 07.00h to 19.00h.

IN-LIFE DATES: From: To:
Route of administration:
oral: gavage
Vehicle:
other: 1.Arachis oil BP 2.Sesame oil 3.corn oil
Details on exposure:
1.Details on exposure
PREPARATION OF DOSING SOLUTIONS: test material soluble in Arachis oil BP.
DIET PREPARATION
- Rate of preparation of diet (frequency):Fresh diets were made and distributed weekly
- 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): Arachis oil BP
- Concentration in vehicle: 0, 30, 300 and 600mg/kg bw/day
- Amount of vehicle (if gavage):
- Lot/batch no. (if required): No data available
- Purity: No data available
2.Details on exposure
PREPARATION OF DOSING SOLUTIONS:
Test material dissolved in Sesame oil

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): Test material dissolved in Sesame oil

- Concentration in vehicle: 0,1000 mg/kg bw/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 materials dissolved in corn oil. The corn oil used i n the dose formulations contained less than 2 mill equivalents of peroxide/kg corn oil.
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): Test material dissolved in corn oil and stable throughout the study.
- Concentration in vehicle: 0, 50,200,400mg/kg bw/day
- Amount of vehicle (if gavage): 2ml/kg bw
- 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. Impregnation procedure: [artificial insemination / purchased timed pregnant / cohoused]
- If cohoused:
- M/F ratio per cage: 1:1
- Length of cohabitation:
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility.
- Further matings after two unsuccessful attempts: [no / yes (explain)]
- Verification of same strain and source of both sexes: [yes / no (explain)]
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy
- Any other deviations from standard protocol:

3. A artificially inseminated female animals were used
Duration of treatment / exposure:
1.
11 week ( (including a two week pre-pairing phase, pairing, gestation and early lactation for females)
2. 9 days (7th - 16th gestation day)
3.13 days ( gestational days 6-19)
Frequency of treatment:
Daily
Duration of test:
11 weeks
Remarks:
Study 1.
0, 30,300,600 mg/kg bw/day
Study 2.1000.0 mg/kg bw/day
No. of animals per sex per dose:
Study 1
Total:80
0 mg/kg bw/day: 10 male and 10 female
30mg/kg bw/day: 10 male and 10 female
300mg/kg bw/day: 10 male and 10 female
600mg/kg bw/day: 10 male and 10 female
Study 2
Total:40
0 mg/kg bw/day:20
1000mg/kg bw/day:20
3.Total:104
0 mg/kg bw/day:26
50mg/kg bw/day:26
200mg/kg bw/day:26
400 mg/kg bw/day:26

Control animals:
yes, concurrent vehicle
Details on study design:
No data available
Maternal examinations:
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 for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes


WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): yes
3.Parental animals observation and examinations
CAGE SIDE OBSERVATIONS: yes

DETAILED CLINICAL OBSERVATIONS: Yes

Time schedule: Animals were observed daily before (gd 0-5). During (gd 6-19). and after (gd 20-30) dosing
For clinical signs of toxicity .

BODY WEIGHT: Yes
Time schedule for examinations: animals were weighed on the mornings of gd 0, 6 through 19,25 and 30
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes Food was weighed on gd 0,3,6,9,12,15,18,19,22,25,28 and 30.
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:

Ovaries and uterine content:
1&3The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: No data
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Other:
Fetal examinations:
1&3- External examinations: Yes: all per litter
- Soft tissue examinations:No data
- Skeletal examinations: No data
- Head examinations: No data
Statistics:
General Linear Models (GLM) procedures were applied for the analyses of variance (ANOVA) of maternal and fetal parameters. Prior to GLM analysis ,an arcsine-square root transformation was performed on a litter- derived percentage data (Snedecor and Cochran. 1967) and Bartletts test for homogeneity of variance was performed on all data to be analysed by ANOVA (Winer. 1962). GLM analysis determined the significance of dose-response relationships and the significance of dose effects. Replicate effects and dose x replicate interactions. When ANOVA revealed a significant (p<0.05) dose effect. Williams' and Dunnett's Multiple Comparison Tests (Williams, 1971: 1972: Dunnett. 1955: 1964) compared CAM-exposed to control groups. One-tailed tests were used for all pairwise comparisons except maternal body and organ weights. Maternal food consumption. fetal body weight and percent males per litter. Non-significant (p>0.05) dose x replicate effects on selected fetal parametric measures were considered justification for pooling data across replicates for nonparametric analysis on related measures. When a significant (p<0.05#) dose x replicate' interaction occurred, the data for that endpoint and f0.r any related nominal scale data were analysed separately for dose effects within each replicate in the study. As well as for all replicates combined. Nominal scale measures were analysed by a x2 test for independence and by a test 'f0.r linear trend on proportions. When x2 test showed significant group differences. a one tailed Fisher's exact probability test was used for pairwise comparisons of CAM and control groups.
Indices:
No data available
Historical control data:
No data available
Clinical signs:
no effects observed
Description (incidence and severity):
1.Assessment of surface righting ability on Day 1; offspring clinical signs did not indicate any effect of maternal treatment
Dermal irritation (if dermal study):
not specified
Mortality:
mortality observed, non-treatment-related
Description (incidence):
3.There were no dose related maternal deaths during the study, but two does were removed from the 400 mg/kg/day dose group because of dosing error.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
1.At 600 mg/kg bw/day initial offspring body weight was similar to control but weight gain to Day 4 was statistically significantly lower than control. Mean offspring body weights, litter weight and weight gains to Day 4 of age were unaffected by maternal treatment at 30 and 300 mg/kg bw/day.
3.Maternal body weight before, during and after treatment period in the treated animals was also comparable to control weights at all gestational ages. Maternal weight gain during the dosing period tended to decrease with increasing dose. Maternal weight gain relative to control animals was reduced 13%, 5% and 59% in the 50, 200 and 400 mg/kg/day dose groups respectively.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
Food consumption in all dose -treated groups was similar to control throughout the study.
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
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
Gravid uterine and absolute and relative maternal liver weights were similar to vehicle control values.
Gross pathological findings:
no effects observed
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
not specified
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:
not specified
Changes in pregnancy duration:
no effects observed
Description (incidence and severity):
1.Gestation length was considered to be unaffected by treatment at 30, 300 and 600 mg/kg bw/day.
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): not specified
Changes in number of pregnant:
not specified
Other effects:
no effects observed
Description (incidence and severity):
2.No maternal toxic effects were observed
Dose descriptor:
NOAEL
Effect level:
> 400 - <= 1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
clinical signs
body weight and weight gain
gross pathology
Remarks on result:
other: No developmental toxic effects observed
Abnormalities:
not specified
Localisation:
not specified
Fetal body weight changes:
no effects observed
Description (incidence and severity):
1.At 600 mg/kg bw/day initial offspring body weight was similar to control but weight gain to Day 4 was statistically significantly lower than control. Mean offspring body weights, litter weight and weight gains to Day 4 of age were unaffected by maternal treatment at 30 and 300 mg/kg bw/day.
3.Average fetal body weight were unaffected
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:
not specified
Changes in sex ratio:
no effects observed
Changes in litter size and weights:
no effects observed
Description (incidence and severity):
3.Live litter size and average fetal body weight were unaffected
Changes in postnatal survival:
not specified
External malformations:
no effects observed
Description (incidence and severity):
Macroscopic necropsy findings did not indicate any effect of treatment at 30, 300 and 600 mg/kg bw/day.
Skeletal malformations:
no effects observed
Visceral malformations:
no effects observed
Other effects:
no effects observed
Description (incidence and severity):
2.No embryonal / fetal toxicity as well as none teratogenicity was observed
3.No incidence of anatomical variation or defects were observed
Dose descriptor:
NOAEL
Effect level:
> 400 - <= 1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
not specified
Basis for effect level:
reduction in number of live offspring
changes in litter size and weights
external malformations
Remarks on result:
other: No developmental toxic effects was 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:
No Observed Adverse Effect Level (NOAEL) for reproductive and developmental toxicity was considered to 600mg/kg/day, When male and female rats were treated with test material orally.
Executive summary:

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

Study 1

The reproductive and developmental toxicity study of test material was performed on male and femaleWistar Han (TM): RCCHan(TM): WIST strain rats. The test material was dissolved in Arachis oil BP and administered in dose concentration 0, 30,300,600mg/kg bw /day via oral gavage for up to eleven weeks (including a two week pre-pairing phase, pairing, gestation and early lactation for females).Each treatment group contain 10 male and 10 female rats while a control group of ten males and ten females was dosed with vehicle alone (Arachis oil BP). Clinical signs, bodyweight change, dietary intake and water consumption were monitored during the study. Pairing of animals within each dose group was undertaken on a one male: one female basis within each treatment group on Day 5 of the study, with females subsequently being allowed to litter and rear their offspring to Day 5 of lactation. An additional pairing for high dose females that failed to achieve pregnancy was performed to fully assess mating performance and fertility. During the lactation phase, daily clinical observations were performed on all surviving offspring, together with litter size and offspring weights and assessment of surface righting reflex. Adult males were terminated on Day 52 of the study following the completion of the second pairing at 600 mg/kg bw/day. Females and offspring were terminated on day 5 post partum. Any female which did not produce a pregnancy (unless allocated to a further mating phase) was terminated on or after Day 25 post coitum. All animals were subjected to a gross necropsy examination and histopathological evaluation of reproductive tissues was performed. Additional male organ weight and detailed histopathological examination of the testes were performed to more fully assess male fertility.

 

There were no unscheduled deaths in the study.Transient post-dosing salivation was observed from Day 7 at 600 mg/kg bw/day and Day 13 at 300 mg/kg bw/day. This sign was observed regularly throughout the treatment period with all animals being affected at both dosages, although the incidence of thisfinding was greatest at the high dosage. Transient post dosing salivation was also observed at 30 mg/kg bw/day for two females on Day 2 and one male on Day 50 of this study.At 600 mg/kg bw/day body weight gain of males was statistically significantly lower than control during thefirst week of treatment. Subsequent body weight gains were considered to reflect normal biological variation, but overall gain at termination remained lower than control.

Bodyweight gain of males at 30 and 300 mg/kg bw/day and for females at all dosages, throughout the pre-pairing, gestation and lactation phases of the study, were considered to have been unaffected by treatment. Food consumption for both sexes was considered to have been unaffected by treatment throughout the study, and including gestation and lactation phases for females, at 30, 300 and 600 mg/kg bw/day. Food efficiency: At 600 mg/kg bw/day food conversation efficiency for males was lower than control during week 1; subsequent food utilisation was similar to control. Food conversation efficiency of males at 30 and 300 mg/kg bw/day and for females at all dosages, throughout the pre-pairing, gestation and lactation phases of the study, were considered to have been unaffected by the treatment. Male reproductive organ weights were unaffected by treatment at 30, 300 and 600 mg/kg bw/day and did not indicate any effect on fertility. Histopathological examinations did not indicate any effect of treatment at 600 mg/kg bw/day and these examinations, including detailed assessment of the spermatogenetic cycle for the testes did not indicate any effect on male fertility. Pre-coital interval and mating evidence at the times of conception did not indicate any adverse effect of treatment on mating performance at 30, 300 and 600 mg/kg bw/day. Fertility: At 600 mg/kg bw/day, only seven females delivered a litter following the initial pairing but subsequent re-mating and additional assessment of male organ weight and detailed testicular histopathology did not indicate any treatment related effect on fertility for either sex. There was also no effect of treatment on fertility at 30 and 300 mg/kg bw/day. Gestation length was considered to be unaffected by treatment at 30, 300 and 600 mg/kg bw/day. Litter responses: Offspring litter size, sex ratio and viability: There was no effect of maternal treatment on corpora lutea and implantations counts, pre- and postimplantation loss, number of offspring born, sex ratio or subsequent survival to Day 4 of age at 30, 300 and 600 mg/kg bw/day. Assessment of surface righting ability on Day 1; offspring clinical signs and necropsyfindings and did not indicate any effect of maternal treatment. Assessment of surface righting ability on Day 1; offspring clinical signs and necropsyfindings and did not indicate any effect of maternal treatment.Macroscopic necropsyfindings did not indicate any effect of treatment at 30, 300 and 600 mg/kgbw/day. HenceNo Observed Adverse Effect Level (NOAEL) for reproductive and developmental toxicity was considered to 600mg/kg/day,When male and femalerats were treated with test material orally for 11weeks.

Study 2.

Developmental toxicity study of test material was performed on female wistar rats according to OECD guideline 414. The test material dissolved in Sesame oil and administered by oral gavage route in dose concentration 0, 1000 mg/kg be /day from 7th - 16th gestation day.20 female /dose group were used. No adverse effects was observed on treated female. Also No fetal toxicity was observed at dose concentration 1000mg/kg bw /day. Hence NOAEL for reproductive toxicity in the P generation rats and the NOAEL for the F1 generation offspring is considered to be 1000mg/kg/d. When female wistar rats were treated with test material orally.

Study 3.

The developmental toxicity study of test materialwas performed in new Zealand white rabbits.104 rabbits were divided as 26/ dose group. The test material dissolved in corn oil and administered in dose volume 2ml /kg in concentration0, 50, 200, 400 mg/kg bw/day by oral gavage routeon gestational day 6 through 19.The designated dose level were based upon Preliminary dose rang – finding study. A artificially inseminated female animals were used and the day of insemination was designated as Gestational day (gd) 0. Animals were observed daily before (gd 0-5),during (gd 6-19) and after (gd 20-30) dosing for clinical signs of toxicity.Animalswere weighed on the mornings of gd 0, 6 through 19,25and 30.Food was weighed on gd 0,3,6,9,12,15,18,19,22,25,28 and 30. At necropsy,The animals were killed with an injection of sodium pentobarbital(Barber Veterinary Supply. Fayetteville NC)in the marginal ear vein on gd 30. Liver and intact uterus was weighed and corpora lutea were counted. Uterine contents were examined to determine the number of implantation sites, resorptions, dead foetuses and live foetuses. Uteri which had no visible implantation sites was stained with ammonium sulphide (10%) to detect very early resorptions.live foetuses were dissected from the uterus and also anesthetized with sodium pentobarbital.The foetuses were weighed, sexed and subjected to external, soft tissue or skeletal examinations.Half of the foetuses were decapitated prior to dissection, the heads were fixed in bouins solution and then examined by free hand sectioning technique . All fetal carcasses (50% without heads) were stained with alcian blue/ alizarin red S and examined for skeletal malformations

 There were no dose related maternal deaths during the study, but two animals were removed from the 400 mg/kg/day dose group because of dosing error. Food consumption in all dose -treated groups was similar to control throughout the study. Maternal body weight before, during and after treatment period in the treated animals was also comparable to control weights at all gestational ages. Maternal weight gain during the dosing period tended to decrease with increasing dose. Maternal weight gain relative to control animals was reduced 13%, 5% and59%in the 50, 200 and 400 mg/kg/day dose groups respectively. Gravid uterine and absolute and relative maternal liver weights were similar to vehicle control values.

Results of the uterine examination revealed that test material was not developmental toxic effects even at the highest dose. No effect was seen for the percent resorption per litter. The percent late fetal deaths per litter , the percent non live implants per litter or the percent adversely affected implants per litter. The proportion of litters with one or more resorption , late fetal deaths, nonlive implants or adversely affected implants were not affected by dose exposure. Live litter size and average fetal body weight were also unaffected .No external , skeletal and visceral malformations were observed . No incidence of anatomical variation or defects were observed. HenceNo Observed Adverse Effect Level (NOAEL) for maternal and foetal toxicity was considered to be 400 mg/kg/day.When femalenew Zealand white rabbits were treated with test material orally.

Thus, based on the data available fortest chemical,No Observed Adverse Effect Level (NOAEL) was considered to be 600 mg /kg bw .When rats or rabbits were treated with test chemical orally. Thus, comparing this value with the criteria of CLP regulationtest chemicalis not likely to classify as reproductive and developmental toxicant.

 

 

 

 

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
600 mg/kg bw/day
Study duration:
chronic
Species:
other: Rats or rabbits
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 for test chemicals were reviewed to determine the developmental toxicity of test chemical. The studies are as mentioned below:

Study 1

The reproductive and developmental toxicity study of test material was performed on male and femaleWistar Han (TM): RCCHan(TM): WIST strain rats. The test material was dissolved in Arachis oil BP and administered in dose concentration 0, 30,300,600mg/kg bw /day via oral gavage for up to eleven weeks (including a two week pre-pairing phase, pairing, gestation and early lactation for females).Each treatment group contain 10 male and 10 female rats while a control group of ten males and ten females was dosed with vehicle alone (Arachis oil BP). Clinical signs, bodyweight change, dietary intake and water consumption were monitored during the study. Pairing of animals within each dose group was undertaken on a one male: one female basis within each treatment group on Day 5 of the study, with females subsequently being allowed to litter and rear their offspring to Day 5 of lactation. An additional pairing for high dose females that failed to achieve pregnancy was performed to fully assess mating performance and fertility. During the lactation phase, daily clinical observations were performed on all surviving offspring, together with litter size and offspring weights and assessment of surface righting reflex. Adult males were terminated on Day 52 of the study following the completion of the second pairing at 600 mg/kg bw/day. Females and offspring were terminated on day 5 post partum. Any female which did not produce a pregnancy (unless allocated to a further mating phase) was terminated on or after Day 25 post coitum. All animals were subjected to a gross necropsy examination and histopathological evaluation of reproductive tissues was performed. Additional male organ weight and detailed histopathological examination of the testes were performed to more fully assess male fertility.

 There were no unscheduled deaths in the study.Transient post-dosing salivation was observed from Day 7 at 600 mg/kg bw/day and Day 13 at 300 mg/kg bw/day. This sign was observed regularly throughout the treatment period with all animals being affected at both dosages, although the incidence of thisfinding was greatest at the high dosage. Transient post dosing salivation was also observed at 30 mg/kg bw/day for two females on Day 2 and one male on Day 50 of this study.At 600 mg/kg bw/day body weight gain of males was statistically significantly lower than control during thefirst week of treatment. Subsequent body weight gains were considered to reflect normal biological variation, but overall gain at termination remained lower than control.

Bodyweight gain of males at 30 and 300 mg/kg bw/day and for females at all dosages, throughout the pre-pairing, gestation and lactation phases of the study, were considered to have been unaffected by treatment. Food consumption for both sexes was considered to have been unaffected by treatment throughout the study, and including gestation and lactation phases for females, at 30, 300 and 600 mg/kg bw/day. Food efficiency: At 600 mg/kg bw/day food conversation efficiency for males was lower than control during week 1; subsequent food utilisation was similar to control. Food conversation efficiency of males at 30 and 300 mg/kg bw/day and for females at all dosages, throughout the pre-pairing, gestation and lactation phases of the study, were considered to have been unaffected by the treatment. Male reproductive organ weights were unaffected by treatment at 30, 300 and 600 mg/kg bw/day and did not indicate any effect on fertility. Histopathological examinations did not indicate any effect of treatment at 600 mg/kg bw/day and these examinations, including detailed assessment of the spermatogenetic cycle for the testes did not indicate any effect on male fertility. Pre-coital interval and mating evidence at the times of conception did not indicate any adverse effect of treatment on mating performance at 30, 300 and 600 mg/kg bw/day. Fertility: At 600 mg/kg bw/day, only seven females delivered a litter following the initial pairing but subsequent re-mating and additional assessment of male organ weight and detailed testicular histopathology did not indicate any treatment related effect on fertility for either sex. There was also no effect of treatment on fertility at 30 and 300 mg/kg bw/day. Gestation length was considered to be unaffected by treatment at 30, 300 and 600 mg/kg bw/day. Litter responses: Offspring litter size, sex ratio and viability: There was no effect of maternal treatment on corpora lutea and implantations counts, pre- and postimplantation loss, number of offspring born, sex ratio or subsequent survival to Day 4 of age at 30, 300 and 600 mg/kg bw/day. Assessment of surface righting ability on Day 1; offspring clinical signs and necropsyfindings and did not indicate any effect of maternal treatment. Assessment of surface righting ability on Day 1; offspring clinical signs and necropsyfindings and did not indicate any effect of maternal treatment.Macroscopic necropsyfindings did not indicate any effect of treatment at 30, 300 and 600 mg/kgbw/day. HenceNo Observed Adverse Effect Level (NOAEL) for reproductive and developmental toxicity was considered to 600mg/kg/day,When male and femalerats were treated with test material orally for 11weeks.

Study 2.

Developmental toxicity study of test material was performed on female wistar rats according to OECD guideline 414. The test material dissolved in Sesame oil and administered by oral gavage route in dose concentration 0, 1000 mg/kg be /day from 7th - 16th gestation day.20 female /dose group were used. No adverse effects was observed on treated female. Also No fetal toxicity was observed at dose concentration 1000mg/kg bw /day. Hence NOAEL for reproductive toxicity in the P generation rats and the NOAEL for the F1 generation offspring is considered to be 1000mg/kg/d. When female wistar rats were treated with test material orally.

Study 3.

The developmental toxicity study of test materialwas performed in new Zealand white rabbits.104 rabbits were divided as 26/ dose group. The test material dissolved in corn oil and administered in dose volume 2ml /kg in concentration0, 50, 200, 400 mg/kg bw/day by oral gavage routeon gestational day 6 through 19.The designated dose level were based upon Preliminary dose rang – finding study. A artificially inseminated female animals were used and the day of insemination was designated as Gestational day (gd) 0. Animals were observed daily before (gd 0-5),during (gd 6-19) and after (gd 20-30) dosing for clinical signs of toxicity.Animalswere weighed on the mornings of gd 0, 6 through 19,25and 30.Food was weighed on gd 0,3,6,9,12,15,18,19,22,25,28 and 30. At necropsy,The animals were killed with an injection of sodium pentobarbital(Barber Veterinary Supply. Fayetteville NC)in the marginal ear vein on gd 30. Liver and intact uterus was weighed and corpora lutea were counted. Uterine contents were examined to determine the number of implantation sites, resorptions, dead foetuses and live foetuses. Uteri which had no visible implantation sites was stained with ammonium sulphide (10%) to detect very early resorptions.live foetuses were dissected from the uterus and also anesthetized with sodium pentobarbital.The foetuses were weighed, sexed and subjected to external, soft tissue or skeletal examinations.Half of the foetuses were decapitated prior to dissection, the heads were fixed in bouins solution and then examined by free hand sectioning technique . All fetal carcasses (50% without heads) were stained with alcian blue/ alizarin red S and examined for skeletal malformations

 There were no dose related maternal deaths during the study, but two animals were removed from the 400 mg/kg/day dose group because of dosing error. Food consumption in all dose -treated groups was similar to control throughout the study. Maternal body weight before, during and after treatment period in the treated animals was also comparable to control weights at all gestational ages. Maternal weight gain during the dosing period tended to decrease with increasing dose. Maternal weight gain relative to control animals was reduced 13%, 5% and59%in the 50, 200 and 400 mg/kg/day dose groups respectively. Gravid uterine and absolute and relative maternal liver weights were similar to vehicle control values.

Results of the uterine examination revealed that test material was not developmental toxic effects even at the highest dose. No effect was seen for the percent resorption per litter. The percent late fetal deaths per litter , the percent non live implants per litter or the percent adversely affected implants per litter. The proportion of litters with one or more resorption , late fetal deaths, nonlive implants or adversely affected implants were not affected by dose exposure. Live litter size and average fetal body weight were also unaffected .No external , skeletal and visceral malformations were observed . No incidence of anatomical variation or defects were observed. HenceNo Observed Adverse Effect Level (NOAEL) for maternal and foetal toxicity was considered to be 400 mg/kg/day.When femalenew Zealand white rabbits were treated with test material orally.

Thus, based on the data available fortest chemical,No Observed Adverse Effect Level (NOAEL) was considered to be 600 mg /kg bw .When rats or rabbits were treated with test chemical orally. Thus, comparing this value with the criteria of CLP regulationtest chemicalis not likely to classify as reproductive and developmental toxicant.

 

 

 

 

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.