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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 was considered to be in range of 30-500mg/kg body weight/day ,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:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Data is from peer reviewed journal
Qualifier:
according to guideline
Guideline:
other: as below
Principles of method if other than guideline:
Toxicity of test material was assessed in pregnant Sprague-Dawley Rats.
GLP compliance:
not specified
Limit test:
no
Species:
rat
Strain:
other: Crl:CD® (SD) IGS BR VAF®/Plus
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Raleigh, NC
- Age at study initiation: 6-8 weeks old
- Weight at study initiation: Females: 213 to 241 g,
- Fasting period before study: No data available
- Housing: Animals were housed individually in cage.
- Diet (e.g. ad libitum): Certified Rodent DietR 5002 (PMI Nutrition International, St. Louis, MO), ad libitum
- Water (e.g. ad libitum): reverse osmosis deionized water (with chlorine added to the processed water as a bacteriostat), ad libitum
- Acclimation period: acclimatizated for short period.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17.7 ± 26.11°C
- Humidity (%): 30% to 70%
- Air changes (per hr):at least 10 changes per hour of 100% fresh air passed through 99.97% HEPA filters.
- Photoperiod (hrs dark / hrs light): 12:12-h light-dark lighting cycle was used

IN-LIFE DATES: From: To: No data available
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
Dosing formulations were prepared daily from bulk materials. Samples from each concentration of the dosing suspensions (first and last days of treatment) were analyzed for test material content.3 mg/ml conc.level was found to be stable for 10 days when stored at ambient conditions and protected from light.
DIET PREPARATION
- Rate of preparation of diet (frequency):Not applicable
- Mixing appropriate amounts with (Type of food): Not applicable
- Storage temperature of food: Not applicable
VEHICLE
- Justification for use and choice of vehicle (if other than water): corn oil
- Concentration in vehicle: 0, 3, 10 and 30 mg/kg/day
- Amount of vehicle (if gavage): 10 ml/kg
- Lot/batch no. (if required): 103K0107 and 074K0025
- Purity: Not available
Details on mating procedure:
- M/F ratio per cage:1:1
- Length of cohabitation:5 days
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility:Not available
- Further matings after two unsuccessful attempts:Not available
- Verification of same strain and source of both sexes:Not available
- Proof of pregnancy: The presence of spermatozoa and/or a copulatory plug in situ was designated as gestational day 0 (GD 0).
- Any other deviations from standard protocol:Not available
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Samples from each concentration of the dosing suspensions (first and last days of treatment) were analyzed for AIM content.No details of analytical methods.
Duration of treatment / exposure:
11 days (gestational days 7 to 17)
Frequency of treatment:
Daily
Details on study schedule:
- Dose selection rationale: Dosages were selected on the basis of range-finding study. dosages of 0, 1.25, 2.5, 5, or 10 mg/kg/day were administered daily to 8 rats/group on GDs 7 to 17. No adverse AIM related clinical signs and food consumption observed. During the postdosage period, the body weight gains remained increased. All rats were pregnant and all caesarean sectioning and litter observations were comparable among the five groups. No fetal gross external alterations were observed. Based on these data, a dosage greater than 10 mg/kg/day AIM was recommended for the definitive developmental toxicity study in rats.
- Rationale for animal assignment (if not random): The healthy, mated female rats, weighing 213 to 241 g, were assigned to four dosage groups, 25 rats/group, using a computer-generated (weight-ordered) randomization procedure based on body weights recorded on initiation of study.
- Rationale for selecting satellite groups:No data avaiable
- Post-exposure recovery period in satellite groups: No data avaiable
- Section schedule rationale (if not random):No data avaiable
Remarks:
Doses / Concentrations:
0, 3, 10, or 30 mg/kg/day
Basis:
actual ingested
No. of animals per sex per dose:
Total: 100
Control(0)-25 female pregnant rats
3 mg/kg/day-25 female pregnant rats
10 mg/kg/day-25 female pregnant rats
30 mg/kg/day-25 female pregnant rats
Control animals:
yes, concurrent vehicle
Details on study design:
No data available
Positive control:
No data available
Parental animals: Observations and examinations:
Clinical signs and cage side observations-
- Time schedule: twice daily
-parameters examined: abnormal, abortions, and premature deliveries before dosage administration and approximately 1 h later
Body weights –
- Time schedule: start of the study and daily during the dosage and postdosage periods.
Feed consumption –GDs 0, 7, 10, 12, 15, 18, and 21.
Oestrous cyclicity (parental animals):
Implantation sites were examined.
Sperm parameters (parental animals):
Not examined
Litter observations:
Litter size and Fetal body weight /litter were examined.
Postmortem examinations (parental animals):
Organ weight and gross pathology were examined.
Postmortem examinations (offspring):
Gross external alterations- Yes.
Soft tissue alterations- Yes,half of the fetuses in each litter.
Skeletal alterations- Yes, half of the fetuses in each litter.
Statistics:
1.Continuous data were analyzed by using Bartlett’s test of homogeneity of variances and the analysis of variance.
2.Dunnett’s test -identify statistical significance of individual groups.
3.Kruskal-Wallis test or Dunn’s method of multiple comparisons-identify statistical significance of individual groups.
4.Greater than 75% ties-Fisher’s exact test
Reproductive indices:
Dead or resorbed conceptuses indice and live male fetuses indice were examined.
Offspring viability indices:
Number of dead fetuses were observed.
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
Occasional incidences of chromorhinorrhea, excess salivation, sparse hair coat, localized alopecia, urine-stained abdominal fur, soft or liquid feces, or decreased activity were observed in some animals from each group of rats.
However, none of these clinical signs were attributed to test material because the number of affected rats was not dosage related and did not significantly differ between control and treated groups.
Dermal irritation (if dermal study):
not specified
Mortality:
no mortality observed
Description (incidence):
No effect was observed on survival of treated female rats as compared to control.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Body weight gains were unaffected by dosages of test material , body weight gains were 99.0%, 99.9%, and 101.2% of the control value over the same time period. Body weight gains were also comparable during the postdosage period (GDs 18 to 21).
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
not examined
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Other effects:
not examined
Reproductive function: oestrous cycle:
not specified
Reproductive function: sperm measures:
not examined
Reproductive performance:
no effects observed
Description (incidence and severity):
Pregnancy occurred in 24 (96%), 25 (100%), 24 (96%), and 21 (84%) of the 25 rats in the 0, 3, 10, and 30 mg/kg/day
dosage groups, an event determined before exposure was initiated. No caesarean sectioning parameters were affected by dosages of test material.Reproductive performance was comparable with that observed in the control group and were within the ranges observed historically.
No significant effect were observed on corpora lutea, implantations, litter sizes, live fetuses, resorptions, percentage of dead or resorbed conceptuses and percentage of live male fetuses of treated female rats as compared to control.
Mortality: No effect was observed on survival of treated female rats as compared to control.

Clinical sing: Chromorhinorrhea, excess salivation, sparse hair coat, localized alopecia, urine-stained abdominal fur, soft or liquid feces, or decreased activities were observed in a few animals from each group of rats.

None of these clinical signs were attributed to treatment because the number of affected rats was not dosage related and did not significantly differ between control and treated groups.

Body weight and body weight gain: No effects were observed on body weight and body weight gain of treated female rat during dosing and post dosage period (GDs 18 to 21).

Food consumption: Absolute feed consumption was 101.0%, 102.1%, and 102.6% of the control value for 3, 10, and 30 mg/kg/day dose group.

No effect was observed on food consumption of treated female rats as compared to control.

Reproductive performenus: No significant effect were observed on corpora lutea, implantations, litter sizes, live fetuses, resorptions, percentage of dead or resorbed conceptuses and percentage of live male fetuses of treated female rats as compared to control.

Gross pathology: No alterations were observed on litters with fetuses and % fetuses/litter in treated female rats as compared to control.
Key result
Dose descriptor:
NOAEL
Effect level:
30 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
female
Basis for effect level:
other: No advease effect on survival, clilnical signs, body weight, feed consumption, gross pathology, reproductive performenus and fetal development
Remarks on result:
other: No effects on reproductive performance
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:
not specified
Dermal irritation (if dermal study):
not specified
Mortality / viability:
no mortality observed
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Fetal body weight did not differ significantly from that of the controls
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 examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Description (incidence and severity):
Gross external alterations of the fetuses made showed a depressed left eye bulge, a fleshy thoracic protrusion, and the presence of an extra hind limb in the control fetus. Skeletal examination of the same control fetus revealed a small left eye socket; the presence of three femurs, tibia, and fibulas on the right side of the body (partially fused), and extra metatarsals, digits, and three sets of phalanges (four, five, and five) on the right hindlimbs.

No soft tissue malformations occurred in any of the groups. Soft tissue variations were limited to folded retinas in 7
fetuses from two different litters at 10 mg/kg/day, slight dilation of the renal pelvis in one fetus from the control and one fetus from the 3 mg/kg/day dosage groups, and an aberrant umbilical artery in one and two fetuses at 3 and 10 mg/kg/day, respectively. Skeletal
malformations were absent, and skeletal variations were limited to infrequent incidences of delayed ossification at various bone ossification centers (non–dose dependent) and the presence of cervical ribs at the 7th cervical vertebra in four fetuses in each of the control and 30 mg/kg/day dosage groups.
Histopathological findings:
no effects observed
Other effects:
not specified
Behaviour (functional findings):
not specified
Developmental immunotoxicity:
not specified
Mortality: No effect was observed on number of live fetuses in treated groups as compared to control.

Fetus weight: No effect was observed on fetus weight in treated groups as compared to control.

Gross pathology: No gross external fetal alterations were observed in treated rats as compared to control.

Soft tissue malformations:
Limited to infrequent incidences of skeletal variations as delayed ossification at various bone ossification centers (non–dose dependent) and the presence of cervical ribs at the 7th cervical vertebra in four foetuses of 30 mg/kg/day and in control.

Folded retinas in seven foetuses and aberrant umbilical artery in two fetuses at 10 mg/kg/day

Slight dilation of the renal pelvis in one fetus at 3 mg/kg/day was observed.

All fetal gross external, soft tissue, or skeletal alterations (malformations or variations) were considered unrelated to AIM
because (1) neither the fetal nor litter incidences were dosage dependent; (2) the incidences did not significantly differ from the control group values; and/or (3) the incidences were within the ranges observed historically at the Testing Facility.
Key result
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
30 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No adverse effect on live and dead fetous, fetal body weight, gross external, soft tissue, skeletal variations and malformations of fetouese
Remarks on result:
other: No developmental 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
Reproductive effects observed:
not specified
Treatment related:
not specified
Relation to other toxic effects:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified
Conclusions:
NOAEL was considered to be 30 mg/kg/day for P0 and F1 generation when Crl:CD® (SD) IGS BR VAF®/Plus female rats treated with test material orally.
Executive summary:

In this study the toxicity of test material was evaluated in Crl:CD® (SD) IGS BR VAF®/Plus female rats during formative stages of pregnancy and its effect on developing embryo-fetus orally by gavage in the concentration of 0, 3, 10 and 30 mg/kg/day. No effects were observed on survival, body weight and body weight gain and food consumption of treated female rats as compared to control. Chromorhinorrhea, excess salivation, sparse hair coat, localized alopecia, urine-stained abdominal fur, soft or liquid feces, or decreased activities were observed in a few animals from each group of rats. None of these clinical signs were attributed to treatment because the number of affected rats was not dosage related and did not significantly differ between control and treated groups. Similarly, No significant effect were observed on corpora lutea, implantations, litter sizes, live fetuses, resorptions, percentage of dead or resorbed conceptuses and percentage of live male fetuses and no alterations were observed on litters with fetuses and % fetuses/litter in treated female rats as compared to control. In addition, no effects were observed on number of live fetuses, fetus weight and on gross external fetal alterations in treated rats as compared to control. Limited to infrequent incidences of skeletal variations as delayed ossification at various bone ossification centers (non–dose dependent) and the presence of cervical ribs at the 7thcervical vertebra in four foetuses were observed in 30 mg/kg/day and in control. Folded retinas in seven foetuses and aberrant umbilical artery in two fetuses at 10 mg/kg/day and Slight dilation of the renal pelvis in one fetus at 3 mg/kg/day was observed. All fetal gross external, soft tissue, or skeletal alterations (malformations or variations) were considered unrelated to treatment because (1) neither the fetal nor litter incidences were dosage dependent; (2) the incidences did not significantly differ from the control group values; and/or (3) the incidences were within the ranges observed historically at the Testing Facility. Therefore, NOAEL was considered to be 30 mg/kg/day for P0 and F1 generation when Crl: CD® (SD) IGS BR VAF®/Plus female rats were treated with test material orally by gavge for 11 days.

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
500 mg/kg bw/day
Study duration:
subacute
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

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

In this study the toxicity of test material was evaluated in Crl:CD® (SD) IGS BR VAF®/Plus female rats during formative stages of pregnancy and its effect on developing embryo-fetus orally by gavage in the concentration of 0, 3, 10 and 30 mg/kg/day. No effects were observed on survival, body weight and body weight gain and food consumption of treated female rats as compared to control. Chromorhinorrhea, excess salivation, sparse hair coat, localized alopecia, urine-stained abdominal fur, soft or liquid feces, or decreased activities were observed in a few animals from each group of rats. None of these clinical signs were attributed to treatment because the number of affected rats was not dosage related and did not significantly differ between control and treated groups. Similarly, No significant effect were observed on corpora lutea, implantations, litter sizes, live fetuses, resorptions, percentage of dead or resorbed conceptuses and percentage of live male fetuses and no alterations were observed on litters with fetuses and % fetuses/litter in treated female rats as compared to control. In addition, no effects were observed on number of live fetuses, fetus weight and on gross external fetal alterations in treated rats as compared to control. Limited to infrequent incidences of skeletal variations as delayed ossification at various bone ossification centers (non–dose dependent) and the presence of cervical ribs at the 7thcervical vertebra in four foetuses were observed in 30 mg/kg/day and in control. Folded retinas in seven foetuses and aberrant umbilical artery in two fetuses at 10 mg/kg/day and Slight dilation of the renal pelvis in one fetus at 3 mg/kg/day was observed. All fetal gross external, soft tissue, or skeletal alterations (malformations or variations) were considered unrelated to treatment because (1) neither the fetal nor litter incidences were dosage dependent; (2) the incidences did not significantly differ from the control group values; and/or (3) the incidences were within the ranges observed historically at the Testing Facility. Therefore, NOAEL was considered to be 30 mg/kg/day for P0 and F1 generation when Crl: CD® (SD) IGS BR VAF®/Plus female rats were treated with test material orally by gavge for 11 days.

Study 2

In a dose range finding study, develpmental toxicity was evaluated in Crl:CD(SD) IGS BR VAF/plus pregnant female rats by using test material in the concentration of  0, 1.25, 2.5, 5 and 10 mg/kg/day orally by gavage in corn oil. No mortality and clinical sign were observed in treated female rats as compared to control. Increase in Body weight gains were observed in treated rats during the entire dosage period, the gestation period following the initiation of dosing and the entire gestation period, but this was not considered to be treatment related. Similarly, No effects were observed on Number and distribution of corpora lutea, implantation sites and uterine contents. No thoracic abdominal and pelvic viscera abnormalites were observed in treated rats as compared to control. In addition, No effect on sex of fetuses and no Gross external alterations were observed in fetuses of treated rats as compared to control. Therefore, NOAEL was considered to be > 10 mg/kg/day for P0 and F1 generation when Crl:CD(SD) IGS BR VAF/plus pregnant female rats were treated with test material orally for 11 days.

Study 3

In a Subchronic oral toxicity study, FDRL male and female rats were treated with test material in the concentration of 0,3.55 mg/kg bw/day for male and 4.10 mg/kg bw/day for female orally in diet. No significant effect in body weight and body weight gain, food consumption and food intake of treated male and female rats were observed as compared to control.Slightly decrease haemoglobin and Decrease mean blood urea nitrogen level was observed in male rats but were not considered as statistically significant as compared to control.In addition, no gross pathological changes, no liver and kidney weight change and No histopathological changes were observed in liver, kidneys, stomach, small and large intestines, spleen, pancreas, heart, lungs, bone marrow, muscle, brain, spinal cord, bladder, adrenals, thyroid, pituitary, gonads, salivary glands and lymph nodes of treated male and femlae rats as compared to control. Therefore,NOAEL was considered to be 3.55 mg/kg bw/day for male and 4.10 mg/kg bw/day for female rats when FDRLmale and female rats were treated with test material orally in diet for 90 days.The study does not conclude higher dose value. Hence it is acceptable to take such low NOAEL .

 

Study 4

 

In a Subchronic Oral Toxicity Study, Sprague-Dawley Crl:CD IGS BR male and female rats were treated with test material in the concentration of 0, 5, 30 and 500 mg/kg/day orally by gavage in corn oil. No effects were observed on survival, clinical sign and Functional/behavioral parameters were observed in treated rats as compared to control. Statistically significant decrease in body weight gains were observed during week 7 and statistically significantly increased in body weight gains during weeks 9 and 13 were observed in 500 mg/kg/day treated rats but, not significant as compared to control at the end of study and considered to be non-toxic. Similarly, no effect s was observed on food consumption, food efficiency, water consumption, Ophthalmoscopic examination and hematological parameters of treated rats as compared to control. Statistically significant increase in albumin, plasma creatinine, total protein and cholesterol and statistically significant decrease in AST and Chloride level were observed in male rats and statistically significant increase in plasma creatinine, total protein and cholesterol and statistically significant decrease in alkaline phosphatase, AST and Bilirubin level of treated female rats at 500 mg/kg/day as compared to control. All individual values were within the normal range for rats of the strain and age used, and the intergroup difference was considered not to be toxicologically significant. In addition, no reproductive orange gross pathological changes were observed in treated rats as compared to control. Some absolute and relative Adrenals, Heart, Kidneys, Liver and Spleen weigh changes were observed in male and female rats as compared to control. All individual values for observed changes were within the normal range for rats of the strain and age used and in the absence of a dose-related response the differences were considered to be of no toxicological importance. At 500 mg/kg/day, minimal to slight adipose infiltration in Bone and bone marrow , minimal to moderate Globular accumulations of eosinophilic material in Kidneys, minimal generalized and centrilobular Hepatocyte enlargement of Liver and minimal Follicular cell hypertrophy of Thyroid were observed in treated male rats and minimal centrilobular Hepatocyte enlargement of Liver in female rats as compared to control. Kidney changes were also observed in 30 mg/kg/day treated male rats. Liver and Kidneys changes were commonly seen following the administration of xenobiotics and is generally regarded as adaptive in nature in the absence of associated degenerative changes. No effects were observed on mammary glands, ovaries, prostate and testes of treated rats as compared to control. Therefore, NOAEL was considered to be 500 mg/kg/day when Sprague-Dawley Crl:CD IGS BR male and female rats were treated with test material orally by gavage for 90 days.

Study 5

The reproductive and developmental toxicity of the test chemical was performed on male and female wistar rats, the test chemical was administered into the rats by oral gavage in the concentrations of 0. 250, 500, 750 and 1000 mg/kg bw/day once on pregnancy day 11.The vehicle used to gavage the test chemical was corn oil. The source for the animals was Center for breeding of laboratory animals of the Oswaldo Cruz Foundation (Cecal-Fiocruz) and weighed around 200 -300 g at the initiation of the study. The rats were kept in the controlled temperature environment wherein the humidity was maintained around 70% and photoperiod was adjusted to 12 hours dark and 12 hours light. The rats were housed in standard plastic cages with stainless steel cover lids and wood shavings as bedding and kept under conditions of controlled temperature. The animals were mated in the ratio of 2:1 (2 females and 1 males) and were cohabited for 2 hours. The pregnancy was confirmed by the presence of sperm in the vaginal smear and was referred to as day '0' of pregnancy. On day 21 of gestation all dams were anesthetized and killed by CO2 inhalation. The gravid uterus was weighed with its contents and the number of implantation sites, living and dead fetuses, and resorptions was recorded. The ratio of resorptions per implantations, and the percentage of resorptions per implantations per litter were substantially increased, while the ratio of live fetuses per implantations per litter was drastically decreased, in dams treated with the highest dose of the test chemical (1000 mg/kg bw/day). In maternal animals, no signs of toxicity was observed till 750 mg/kg bw/day. The animals gained normal weight until sacrifice. After sacrifice and gross necropsy, the gravid uterus weight did not differ significantly till 750 mg/kg bw/day. Although, at 1000 mg/kg bw/day dose group, the number of resorptions were observed to be increased in maternal animals. Also, the gravid uterine weight was observed to be decreased significantly as compared to the control group. Also, the ratio of implantation sites to resorptions was found to be lower as compared to the control group. No externally-visible anomaly was noted in fetuses from dams treated orally with test material on pregnancy day 11. Thus, NOAEL was considered to be 750 mg/kg bw/day and LOAEL was considered to be 1000 mg/kg bw/day. When maie and female rats were treated with test material orally once on pregnancy day 11.

Based on the data available from different studies ,test material did not showedreproductive toxicityat dose concentration 500 mg/kg body weight/day,when male and female rats were treated with test material orally,Thus, comparing this value with the criteria ofCLP regulationtest materialis not likely to classify as reproductive toxicant.

 

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, NOAEL for test chemical was considered to be 30mg /kg bw/day .When rats 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:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Data is from peer reviewed journal
Qualifier:
according to guideline
Guideline:
other: as below
Principles of method if other than guideline:
Test material was evaluated for developmental toxicity in presumed pregnant Sprague-Dawley rats.
GLP compliance:
not specified
Limit test:
no
Species:
rat
Strain:
other: Crl:CD® (SD) IGS BR VAF®/Plus
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Raleigh, NC
- Age at study initiation: 6-8 weeks old
- Weight at study initiation: Females: 213 to 241 g,
- Fasting period before study: No data available
- Housing: Animals were housed individually in cage.
- Diet (e.g. ad libitum): Certified Rodent DietR 5002 (PMI Nutrition International, St. Louis, MO), ad libitum
- Water (e.g. ad libitum): reverse osmosis deionized water (with chlorine added to the processed water as a bacteriostat), ad libitum
- Acclimation period: acclimatizated for short period.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17.7 ± 26.11°C
- Humidity (%): 30% to 70%
- Air changes (per hr):at least 10 changes per hour of 100% fresh air passed through 99.97% HEPA filters.
- Photoperiod (hrs dark / hrs light): 12:12-h light-dark lighting cycle was used

IN-LIFE DATES: From: To: No data available
Route of administration:
oral: gavage
Type of inhalation exposure (if applicable):
not specified
Vehicle:
corn oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
Dosing formulations were prepared daily from bulk materials. Samples from each concentration of the dosing suspensions (first and last days of treatment) were analyzed for AIM content.3 mg/ml conc.level was found to be stable for 10 days when stored at ambient conditions and protected from light.
DIET PREPARATION
- Rate of preparation of diet (frequency):Not applicable
- Mixing appropriate amounts with (Type of food): Not applicable
- Storage temperature of food: Not applicable
VEHICLE
- Justification for use and choice of vehicle (if other than water): corn oil
- Concentration in vehicle: 0, 3, 10 and 30 mg/kg/day
- Amount of vehicle (if gavage): 10 ml/kg
- Lot/batch no. (if required): 103K0107 and 074K0025
- Purity: Not available
Analytical verification of doses or concentrations:
not specified
Details on mating procedure:
- M/F ratio per cage:1:1
- Length of cohabitation:5 days
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility:Not available
- Further matings after two unsuccessful attempts:Not available
- Verification of same strain and source of both sexes:Not available
- Proof of pregnancy: The presence of spermatozoa and/or a copulatory plug in situ was designated as gestational day 0 (GD 0).
- Any other deviations from standard protocol:Not available

Duration of treatment / exposure:
11 days (gestational days 7 to 17)
Frequency of treatment:
Daily
Duration of test:
gestational day 21
Remarks:
Doses / Concentrations:
0, 3, 10, or 30 mg/kg/day
Basis:
no data
No. of animals per sex per dose:
Total:100
Control(0)-25 female pregnant rats
3 mg/kg/day-25 female pregnant rats
10 mg/kg/day-25 female pregnant rats
30 mg/kg/day-25 female pregnant rats
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Dosages were selected on the basis of range-finding study. dosages of 0, 1.25, 2.5, 5, or 10 mg/kg/day were administered
daily to 8 rats/group on GDs 7 to 17. No adverse AIM related clinical signs and food consumption observed. During the postdosage period, the
body weight gains remained increased. All rats were pregnant and all caesarean sectioning and litter observations were comparable among the five groups. No fetal gross external alterations were observed. Based on these data, a dosage greater than 10 mg/kg/day AIM was recommended for the definitive developmental toxicity study in rats.
- Rationale for animal assignment (if not random): The healthy, mated female rats, weighing 213 to 241 g, were assigned to four dosage groups, 25 rats/group, using a computer-generated (weight-ordered) randomization procedure based on body weights recorded on initiation of study.
- Rationale for selecting satellite groups:No data avaiable
- Post-exposure recovery period in satellite groups: No data avaiable
- Section schedule rationale (if not random):No data avaiable
Maternal examinations:
CAGE SIDE OBSERVATIONS: yes, survival observed
DETAILED CLINICAL OBSERVATIONS: Yes
BODY WEIGHT: Yes
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data
POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day: On gestational day 21.
OTHER:abortions and premature deliveries-Yes, Organ weight and gross pathology were examined.
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
Fetal examinations:
- External examinations: Yes
- Soft tissue examinations: Yes
- Skeletal examinations: Yes
- Head examinations: No data
Statistics:
Continuous data were analyzed by using Bartlett’s test of homogeneity of variances and the analysis of variance. Dunnett’s test was used to identify statistical significance of individual groups. If the analysis of variance was not appropriate, the Kruskal-Wallis test or Dunn’s method of multiple comparisons was used to identify the statistical significance of the individual groups. If there were greater than 75% ties, Fisher’s exact test was used to analyze the data.
Indices:
Dead or resorbed conceptuses indice and live male fetuses indice were examined.
Historical control data:
No data avaialble
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
Chromorhinorrhea, excess salivation, sparse hair coat, localized alopecia, urine-stained abdominal fur, soft or liquid feces, or decreased activities were observed in a few animals from each group of rats. None of these clinical signs were attributed to treatment because the number of affected rats was not dosage related and did not significantly differ between control and treated groups.
Dermal irritation (if dermal study):
not specified
Mortality:
no mortality observed
Description (incidence):
No effect was observed on survival of treated female rats as compared to control.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No effects were observed on body weight and body weight gain of treated female rat during dosing and post dosage period (GDs 18 to 21).
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
Absolute feed consumption was 101.0%, 102.1%, and 102.6% of the control value for 3, 10, and 30 mg/kg/day dose group.
No effect was observed on food consumption of treated female rats as compared to control.
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:
not specified
Gross pathological findings:
no effects observed
Description (incidence and severity):
No alterations were observed on litters with fetuses and % fetuses/litter in treated female rats as compared to control.
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:
not specified
Total litter losses by resorption:
no effects observed
Description (incidence and severity):
No significant effect were observed litter sizes, live fetuses, resorptions
Early or late resorptions:
no effects observed
Description (incidence and severity):
No significant effect were observed
Dead fetuses:
no effects observed
Description (incidence and severity):
No significant effect were observed percentage of live male fetuses of treated female rats as compared to control.
Changes in pregnancy duration:
not specified
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): no effects observed
Changes in number of pregnant:
not specified
Other effects:
not specified
Details on maternal toxic effects:
Maternal toxic effects:no effects

Details on maternal toxic effects:
Mortality: No effect was observed on survival of treated female rats as compared to control.

Clinical sing: Chromorhinorrhea, excess salivation, sparse hair coat, localized alopecia, urine-stained abdominal fur, soft or liquid feces, or decreased activities were observed in a few animals from each group of rats.

None of these clinical signs were attributed to treatment because the number of affected rats was not dosage related and did not significantly differ between control and treated groups.

Body weight and body weight gain: No effects were observed on body weight and body weight gain of treated female rat during dosing and post dosage period (GDs 18 to 21).

Food consumption: Absolute feed consumption was 101.0%, 102.1%, and 102.6% of the control value for 3, 10, and 30 mg/kg/day dose group.

No effect was observed on food consumption of treated female rats as compared to control.

Reproductive performence: No significant effect were observed on corpora lutea, implantations, litter sizes, live fetuses, resorptions, percentage of dead or resorbed conceptuses and percentage of live male fetuses of treated female rats as compared to control.

Gross pathology: No alterations were observed on litters with fetuses and % fetuses/litter in treated female rats as compared to control.
Key result
Dose descriptor:
NOAEL
Effect level:
>= 30 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
body weight and weight gain
clinical signs
food efficiency
gross pathology
maternal abnormalities
mortality
Remarks on result:
other: No toxic effects were observed
Abnormalities:
not specified
Localisation:
not specified
Fetal body weight changes:
no effects observed
Description (incidence and severity):
No effect was observed on fetus weight in treated groups as compared to control.
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): no effects observed
Field "Description (incidence and severity)" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.DescriptionIncidenceAndSeverityFetalPupBodyWeightChanges): No effect was observed on fetus weight in treated groups as compared to control.
Reduction in number of live offspring:
no effects observed
Description (incidence and severity):
No effect was observed on number of live fetuses in treated groups as compared to control.
Changes in sex ratio:
not specified
Changes in litter size and weights:
not specified
Changes in postnatal survival:
not specified
External malformations:
not specified
Skeletal malformations:
effects observed, treatment-related
Description (incidence and severity):
Limited to infrequent incidences of skeletal variations as delayed ossification at various bone ossification centers (non–dose dependent) and the presence of cervical ribs at the 7th cervical vertebra in four foetuses of 30 mg/kg/day and in control.
Visceral malformations:
not specified
Other effects:
not specified
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
Mortality: No effect was observed on number of live fetuses in treated groups as compared to control.

Fetus weight: No effect was observed on fetus weight in treated groups as compared to control.

Gross pathology: No gross external fetal alterations were observed in treated rats as compared to control.

Soft tissue malformations:
Limited to infrequent incidences of skeletal variations as delayed ossification at various bone ossification centers (non–dose dependent) and the presence of cervical ribs at the 7th cervical vertebra in four foetuses of 30 mg/kg/day and in control.

Folded retinas in seven foetuses and aberrant umbilical artery in two fetuses at 10 mg/kg/day

Slight dilation of the renal pelvis in one fetus at 3 mg/kg/day was observed.

All fetal gross external, soft tissue, or skeletal alterations (malformations or variations) were considered unrelated to test material
because (1) neither the fetal nor litter incidences were dosage dependent; (2) the incidences did not significantly differ from the control group values; and/or (3) the incidences were within the ranges observed historically at the Testing Facility.
Dose descriptor:
NOAEL
Effect level:
>= 30 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
reduction in number of live offspring
fetal/pup body weight changes
changes in litter size and weights
skeletal malformations
Remarks on result:
other: All fetal gross external, soft tissue, or skeletal alterations (malformations or variations) were considered unrelated to test material
Key result
Abnormalities:
no effects observed
Localisation:
other: not specified
Developmental effects observed:
not specified
Treatment related:
not specified
Relation to maternal toxicity:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified
Conclusions:
NOAEL was considered to be 30 mg/kg/day for F0 and F1 generation when Crl:CD® (SD) IGS BR VAF®/Plus female rats treated with test material.
Executive summary:

Developmental study was initiated to study the activity of test material . In this study, Crl:CD® (SD) IGS BR VAF®/Plus male and female rats were mated for 5 days and the presence of spermatozoa and/or a copulatory plug in situ was designated as gestational day 0 (GD 0). The test material in a corn oil vehicle was administered orally via gavage to four groups of presumed pregnant rats on GDs 7 to 17 at dosages of 0, 3, 10, or 30 mg/kg/day. No effects were observed on survival, body weight and body weight gain and food consumption of treated female rats as compared to control. Chromorhinorrhea, excess salivation, sparse hair coat, localized alopecia, urine-stained abdominal fur, soft or liquid feces, or decreased activities were observed in a few animals from each group of rats. None of these clinical signs were attributed to treatment because the number of affected rats was not dosage related and did not significantly differ between control and treated groups. Similarly, No significant effect were observed on corpora lutea, implantations, litter sizes, live fetuses, resorptions, percentage of dead or resorbed conceptuses and percentage of live male fetuses and no alterations were observed on litters with fetuses and % fetuses/litter in treated female rats as compared to control. In addition, no effects were observed on number of live fetuses, fetus weight and on gross external fetal alterations in treated rats as compared to control. Limited to infrequent incidences of skeletal variations as delayed ossification at various bone ossification centers (non–dose dependent) and the presence of cervical ribs at the 7thcervical vertebra in four foetuses were observed in 30 mg/kg/day and in control. Folded retinas in seven foetuses and aberrant umbilical artery in two fetuses at 10 mg/kg/day and Slight dilation of the renal pelvis in one fetus at 3 mg/kg/day was observed. All fetal gross external, soft tissue, or skeletal alterations (malformations or variations) were considered unrelated to treatment because (1) neither the fetal nor litter incidences were dosage dependent; (2) the incidences did not significantly differ from the control group values; and/or (3) the incidences were within the ranges observed historically at the Testing Facility. Therefore, NOAEL was considered to be 30 mg/kg/day for F0 and F1 generation when Crl: CD® (SD) IGS BR VAF®/Plus female rats were treated with test material orally by gavge for 11 days.

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

Developmental toxicity study

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

Study 1

Developmental study was initiated to study the activity of test material . In this study, Crl:CD® (SD) IGS BR VAF®/Plus male and female rats were mated for 5 days and the presence of spermatozoa and/or a copulatory plug in situ was designated as gestational day 0 (GD 0). The test material in a corn oil vehicle was administered orally via gavage to four groups of presumed pregnant rats on GDs 7 to 17 at dosages of 0, 3, 10, or 30 mg/kg/day. No effects were observed on survival, body weight and body weight gain and food consumption of treated female rats as compared to control. Chromorhinorrhea, excess salivation, sparse hair coat, localized alopecia, urine-stained abdominal fur, soft or liquid feces, or decreased activities were observed in a few animals from each group of rats. None of these clinical signs were attributed to treatment because the number of affected rats was not dosage related and did not significantly differ between control and treated groups. Similarly, No significant effect were observed on corpora lutea, implantations, litter sizes, live fetuses, resorptions, percentage of dead or resorbed conceptuses and percentage of live male fetuses and no alterations were observed on litters with fetuses and % fetuses/litter in treated female rats as compared to control. In addition, no effects were observed on number of live fetuses, fetus weight and on gross external fetal alterations in treated rats as compared to control. Limited to infrequent incidences of skeletal variations as delayed ossification at various bone ossification centers (non–dose dependent) and the presence of cervical ribs at the 7thcervical vertebra in four foetuses were observed in 30 mg/kg/day and in control. Folded retinas in seven foetuses and aberrant umbilical artery in two fetuses at 10 mg/kg/day and Slight dilation of the renal pelvis in one fetus at 3 mg/kg/day was observed. All fetal gross external, soft tissue, or skeletal alterations (malformations or variations) were considered unrelated to treatment because (1) neither the fetal nor litter incidences were dosage dependent; (2) the incidences did not significantly differ from the control group values; and/or (3) the incidences were within the ranges observed historically at the Testing Facility. Therefore, NOAEL was considered to be 30 mg/kg/day for F0 and F1 generation when Crl: CD® (SD) IGS BR VAF®/Plus female rats were treated with test material orally by gavge for 11 days.

Study 2

In a dose range finding study, develpmental toxicity of test material were evaluated in Crl:CD(SD) IGS BR VAF/plus pregnant female rats in the concentration of  0, 1.25, 2.5, 5 and 10 mg/kg/day in corn oil. No mortality and clinical sign were observed in treated female rats as compared to control.Increase in Body weight gains were observed in treated rats during the entire dosage period, the gestation period following the initiation of dosing and the entire gestation period, but this was not considered to be treatment related. Similarly,No effects were observed on Number and distribution of corpora lutea, implantation sites and uterine contents.No thoracic abdominal and pelvic viscera abnormalites were observed in treated rats as compared to control. In addition, No effect on sex of fetuses and no Gross external alterations were observed in fetuses of treated rats as compared to control. Therefore, NOAEL was considered to be > 10 mg/kg/day for P0 and F1 generation when Crl:CD(SD) IGS BR VAF/plus pregnant female rats were treated with test material orally for 11 days.

Study 3

In a teratogenicity study, Wistar female rats were treated with test material in the concentration of 250, 500, 750 and 1000 mg/kg body weight orally by gavage in corn oil at day 11 of pregnancy. No overt signs of maternal toxicity and effect on body weight gain were observed in treated rats as compared to control. Substantially increased in ratio of resorptions per implantations, and the percentage of resorptions per implantations per litter, while the ratio of live fetuses per implantations per litter was drastically decreased at 1000 mg/kg bw in treated rats as compared to control. Similarly, Decreased in gravid uterus weight were observed in dams treated with 1000 mg/kg/day. In addition, no significant effect on externally-visible anomaly and skull defects were observed in fetuses of treated rats as compared to control. Therefore, NOAEL was considered to be 750 ma/kg bw for p generation and 1000 mg/kg bw for F1 generation when Wistar female rats were treated with test material once orally by gavage in corn oil at day 11 of pregnancy.

Thus, based on the data available fortest chemical,No Observed Adverse Effect Level (NOAEL) was considered to be 30mg /kg bw .When rats 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 material is not likely to classify as reproductive and developmental toxicant.

Additional information