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Toxicological information

Developmental toxicity / teratogenicity

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Endpoint:
developmental toxicity
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Data is based on weight of evidence approach prepared from various publication.
Cross-referenceopen allclose all
Reason / purpose:
read-across source
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:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Justification for type of information:
Data is from NTP report
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Principles of method if other than guideline:
Developmental Toxicity Evaluation of test chemical by adinistration of test chemical by Gavage to Swiss CCD-I (trade name)) Mice on Gestational
Days 6 through 17.
GLP compliance:
no
Limit test:
no
Specific details on test material used for the study:
- Molecular formula (if other than submission substance): C7H10N2O2
Species:
mouse
Strain:
other: Crl:CD-le(ICR) BR VAF/Plus
Remarks:
outbred albino Swiss mice
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River laboratories, Inc. (Raleigh, NC).
- Age at study initiation: Females (8-10 weeks); males (10-12 weeks
- Weight at study initiation: Weight Range for Females: 20-35 g on gestation day 0 (day of vaginal plug detection
- Fasting period before study: No data
- Housing: Solid bottom polycarbonate cages with stainless steel wire lids (Laboratory Products, Rochelle Park, NJ).
- Use of restrainers for preventing ingestion (if dermal):no data
- Diet (e.g. ad libitum): Purina ·Certified Rodent Chowe (15002) (Ralston Purina Co., St. Louis, Missouri) available ad libitum to males and females (pelleted chow)
- Water (e.g. ad libitum): deionized/filtered water provided ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 69-75 F
- Humidity (%): 30-70% relative humidity
- Air changes (per hr): 12-14 air changes per hour
- Photoperiod (hrs dark / hrs light):12/12hrs of light and dark

Route of administration:
oral: gavage
Vehicle:
other: Distiiled water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: Test chemical will be
dissolved in distilled water to a concentration determined by the following
formula:
concentration (mg/ml) = dose level (mg/kg)/ dose volume (10.0 ml/kg)
Test chemical will be weighed into a volumetric flask and distilled water will be added to slightly below the mark. test chemical will be dispersed into a visually uniform solution and the flask will then be filled to the mark and agitated a second time. The solution will be sonicated, if necessary, to dissolve the chemical. 'Each concentration of test chemical will be formulated independently once during each' study replicate in a quantity sufficient for use across the entire dosing period for each replicate.
- Storage condition of formulation: Dose formulations will be stored at refrigeratea temperatures in 60 ml amber vials

Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Prior to administration and after administration, each formulation were analyzed to ensure a concentration between 90 and 110% of the target concentration.
Details on mating procedure:
Monogamous breeding pairs were cohabited over night in the home cage of the male. On the following morning, the female was examined for a copulation plug. The day of plug detection was designated as gd 0. Plug-negative females was returned to the males cage and checked for plugs on successive mornings until insemina tion occurs or the treatment groups are filled, whichever comes first.
Duration of treatment / exposure:
11 days (from gestation period day 6 to 17)
Frequency of treatment:
daily
Duration of test:
17 days (examination of fetuses were performed on day 17)
Remarks:
0, 30, 60, 120, and 180 mg/kg/day
No. of animals per sex per dose:
20 pregnant mice in each group
Control animals:
yes, concurrent vehicle
Details on study design:
Administration by gavage on the mornings of gd 6 through 17, using an 18-gauge 1.5 inch stainless steel dosing tube, dose volume was 10ml/kg. On gestational day 17, approximately one to one and a half days before expected parturition. pregnant dams were anesthetized by carbon dioxide gas and sacrificed by cervical dislocation. The uterus and right ovary were removed and fetus examination were performed
Maternal examinations:
Feed comsumption, water consumption, Body weight, liver, and uterus of each female were recorded on the mornings of gd 0, 3, 6-17 and 1mmediately following sacrifice on GD 17.
Ovaries and uterine content:
The uterus and right ovary were removed and uterine contents examined to determine the number of implantation sites, resorptions, dead fetuses, and live fetuses.
Fetal examinations:
fetal examination were performed including head, external and visceral examination.
Statistics:
Arcsine, Square root transformation, Bartletts test, ANOVA and chi- square test were used for statistical analysis
Indices:
Implantation Index and Resorption Index
Historical control data:
No Data Available
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Mouth bleeding before dosing, Cyst on right ovary,Left ovary enclosed in blood· filled cyst were observed at 30mg/kg group; Cyst on left ovary was observed in 60mg/kg group; Uterus enlarged, Rough coat,Vaginal bleeding,Cyst on right ovary were observed in 120mg/kg group females
perioral wetness, Vaginal discharge (brownish red), Lethargic,Convulsing, Cyst on right ovary, Curling hind limbs under bodywere obseved in treated females but this effects were non significant compared to control female mice.
Dermal irritation (if dermal study):
not specified
Mortality:
no mortality observed
Description (incidence):
No mortality were observed at 30, 60, 120 and 180mg/kg dose groups
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No significant change in body weight were observed at 30, 60, 120 and 180mg/kg dose groups compared to control
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
No significant change in feed consumption were observed at 30, 60, 120 and 180mg/kg dose groups compared to control
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
no effects observed
Description (incidence and severity):
No significant change in water consumption were observed at 30, 60, 120 and 180mg/kg dose groups compared to control
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:
effects observed, treatment-related
Description (incidence and severity):
Significant change in liiver and gravid uterus were observed in 180mg/kg group animals
Gross pathological findings:
not specified
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:
not specified
Early or late resorptions:
not specified
Dead fetuses:
not specified
Changes in pregnancy duration:
not specified
Changes in number of pregnant:
not specified
Other effects:
not specified
Dose descriptor:
NOAEL
Effect level:
180 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
mortality
body weight and weight gain
food consumption and compound intake
water consumption and compound intake
organ weights and organ / body weight ratios
Remarks on result:
other: Not Specified
Abnormalities:
not specified
Fetal body weight changes:
not specified
Reduction in number of live offspring:
not specified
Changes in sex ratio:
not specified
Changes in litter size and weights:
not specified
Changes in postnatal survival:
not specified
External malformations:
effects observed, treatment-related
Description (incidence and severity):
Cleft Palate, Ectocardia, Gastroschisis and Short Tail were obseved in 2 fetuses
Skeletal malformations:
effects observed, treatment-related
Description (incidence and severity):
Asymmetrical Fusion of Sternal Plate, Cleft Sternum, Perforated Sternum, Fused Rib Cartilage, Incomplete Ossification, Cartilage Present, Extra Ossification Site(s), Hematoma: Head, Face, Jaw, Neck, Abdomen, Thorax, Hindlimb were observed
Visceral malformations:
effects observed, treatment-related
Description (incidence and severity):
Situs Inversus Viscerum and left hydronephrosis noted in treated animals
Other effects:
not specified
Details on embryotoxic / teratogenic effects:
NO Data Available
Dose descriptor:
LOAEL
Effect level:
180 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
external malformations
skeletal malformations
visceral malformations
Remarks on result:
other: Not Specified
Abnormalities:
not specified
Developmental effects observed:
not specified
Treatment related:
not specified
Conclusions:
The NOAEL value from the maternal observations was considered to be 180mg/kg/day howeveer the LOAEL value from the fetuses observation considered to be 180mg/kg.
Executive summary:

In the present study, developmental Toxicity of test chemical was evaluated by administration of test chemical by Gavage to Swiss CCD-I (trade name) Mice on Gestational Days 6 through 17. The dose levels of 0, 30, 60, 120, 180 mg/kg were selected and administered in pregnant female mice at day 6 to 17 of gestation period. Dams were observed for clinical signs, change in body weight, feed and water consumption and organ weight change specially liver and gravid uterus. The fetuses were observed for external, visceral and skeletal abnormalities. The results of the study revealed clinical toxicity, Cyst on right ovary,Left ovary enclosed in blood· filled cyst  were observed at 30mg/kg group; Cyst on left ovary was observed in 60mg/kg group; Uterus enlarged, Rough coat,Vaginal bleeding,Cyst on right ovary were observed in 120mg/kg group females perioral wetness, Vaginal discharge (brownish red), Lethargic,Convulsing, Cyst on right ovary, Curling hind limbs under body were obseved in treated females but this effects were non significant compared to control female mice but no change in body weight change, feed consumption and water consumption were observed maternal animals at 30,60,120 and 180mg/kg groups. The fetuses observed for external , visceral and skeletal abnormalities, the results were Cleft Palate, Ectocardia, Gastroschisis and Short Tail were obseved in 2 fetuses. Skeletal malformations were observed such as asymmetrical Fusion of Sternal Plate, Cleft Sternum, Perforated Sternum, Fused Rib Cartilage, Incomplete Ossification, Cartilage Present, Extra Ossification Site(s), Hematoma: Head, Face, Jaw, Neck, Abdomen, Thorax, Hindlimb. the visceral abnormalities also obseved in treated animals including, Situs Inversus Viscerum and left hydronephrosis. From the observations and results of the study, The NOAEL value from the maternal observations was considered to be 180mg/kg/day however the LOAEL value from the fetuses observation considered to be 180mg/kg.

Reason / purpose:
read-across source
Reference
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
secondary literature
Justification for type of information:
Data is from HPV document
Qualifier:
according to
Guideline:
other: Combined Repeated, Reproductive and Developmental Toxicity Screening Test
Principles of method if other than guideline:
The above experiment was performed to assess and evaluate the effects of the test chemical on the reproductive and developmental toxicity parameters of the test animals.
GLP compliance:
no
Specific details on test material used for the study:
- Substance type: Organic
- Physical state: Solid
- Analytical purity: 99.7%
Species:
rat
Strain:
other: Crl:WI(Glx/BRL/Han)BR
Details on test animals and environmental conditions:
At study initiation the rats were 13 to 15 weeks old.
Route of administration:
oral: gavage
Vehicle:
not specified
Details on exposure:
Dose volume was 5mL/kg.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Achieved concentrations were 90 to 100% of expected when measured on Week 1 and Week 8
Details on mating procedure:
No data
- Impregnation procedure: cohoused
- M/F ratio per cage: 1/1 of same treatment group
- Length of cohabitation: 15 days
- Proof of pregnancy: Mating was confirmed by the presence of a vaginal plug in situ or by sperm in a vaginal washing. The day on which mating was confirmed was designated Day 0 of gestation
Duration of treatment / exposure:
Males - 2 weeks before pairing, throughout pairing and until the day before necropsy (at least 4 weeks of treatment)
Females- 2 weeks before pairing, throughout pairing and until Day 4 postpartum, inclusive
Frequency of treatment:
Daily
Duration of test:
Approximately 8 weeks
Remarks:
Doses / Concentrations:
0, 70, 155, 350 mg/kg bw/day
Basis:
no data
No. of animals per sex per dose:
10 animals/sex/dose
Control animals:
other: yes, 5% v/v ethanol in corn oil
Details on study design:
No Data Available
Maternal examinations:
one 350mg/kg female, on days 4 and 5 of dosing, showed post-dosing signs of subdued behaviour, twitching, semi-closed eyes, piloerection, was cold and showed staggered movement.
Ovaries and uterine content:
The uterus was examined for number of implantations and resorptions.
Fetal examinations:
Litter observations were conducted for each litter to Day 4 post-partum. Observations recorded number of pups born (live and dead); daily live litter size and sex(reported on Days 1 and 4); daily clinical observations; individual pup weights on Days 1, and 4 post-partum and necropsy findings of dead and culled pups where condition permitted. All offspring were given a macroscopic external and internal examination, then discarded.
Statistics:
Body weight gains, necropsy body weights, food consumption, haematology, clinical chemistry, locomotor and functional observation data were analysed using one way analysis of variance (ANOVA), used separately for each sex. Levene’s test was used to test for equality of variances between groups. Where Levene’s test was not significant( P>O.Ol) comparisons between each treated group and control were made using Dunnett’s test. Regression analysis for dose response was also performed.
Non-parametric methods w ere used in place of the one-way ANOVA for clinical chemistry parameters w ith values above or below the limit of the assay. The non-parametric methods employed w ere the Kruskal-Wallis ANOVA, the Terpstra-Jonckheere test for a dose related trend and the Wilcoxon rank sum test for pairwise comparisons. The numbers of implantation sites and pups born, the percentage of male pups on Day 1 and the mean pup weights were also analysed using non-parametric methods.
Organ weights were analysed using analysis of covariance (ANOVA) and Dunnett’s test, for each sex separately, using the necropsy body weight as covariate.
The gestation index and male and female fertility and fecundity indices were analysed using the Cochran-Armitage trend test and Fisher’s exact test (one sided risk) for pairwise comparisons with control.
Indices:
Implantation Index, Resorption Index
Historical control data:
No Data Available
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
One high dose 350mg/kg group female, on Days 4 and 5 of dosing, showed post-dosing signs of subdued behaviour, twitching, semi-closed eyes, piloerection, was cold and showed staggered movement.





Dermal irritation (if dermal study):
not specified
Mortality:
no mortality observed
Description (incidence):
No adult animals died in the study at dose 70, 155 and 350mg/kg
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
The majority of males in the high dose 350mg/kg group lost weight during the first week of dosing. Although mean body weights increased every week after that, the high dose males were approximately 9% lighter than control males by the end of the study.
Group mean body weight gain was lower in 350mg/kg dose group females than controls during gestation. 155 and 350mg/kg dose females had lower mean body weight gain than controls during lactation.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
At 350mg/kg dose, males and females had reduced food intake compared to controls during pre-pairing. Mean food intakes for females of same group were slightly lower than controls during gestation
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):
no effects observed
Description (incidence and severity):
Functional observation battery: Forelimb grip strength in males during Week 7 showed a dose-related decrease. There were no other dose-related observations on functional effects.
Locomotor activity: No effects
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
Organ weight effects observed in kidneys, adrenal glands and liver. Adjusted mean kidney weight was increased in high dose 350mg/kg males Adjusted mean adrenal weight was increased in high dose females( P<0.05). Adjusted mean liver weight was increased in all treated male groups at 70, 155 and 350mg/kg groups
Gross pathological findings:
no effects observed
Description (incidence and severity):
Necropsy: No necropsy findings that were deemed to be treatment-related.
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Histopathology:There were treatment-related findings in the liver, kidney and lungs. Centrilobular hypertrophy was recorded in the majority of the high 350mg/kg dose male and female livers. In the kidney there was an increase in the level of hyaline droplets in males at 350mg/kg dose and In the lungs there was a minor increase in the level of foamy histiocytes in males and females from 350mg/kg dose group.
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Details on results:
One high dose 350mg/kg group female, on Days 4 and 5 of dosing, showed post-dosing signs of subdued behaviour, twitching, semi-closed eyes, piloerection, was cold and showed staggered movement. No adult animals died in the study at dose 70, 155 and 350mg/kg. The majority of males in the high dose 350mg/kg group lost weight during the first week of dosing. Although mean body weights increased every week after that, the high dose males were approximately 9% lighter than control males by the end of the study.Group mean body weight gain was lower in 350mg/kg dose group females than controls during gestation. 155 and 350mg/kg dose females had lower mean body weight gain than controls during lactation. At 350mg/kg dose, males and females had reduced food intake compared to controls during pre-pairing. Mean food intakes for females of same group were slightly lower than controls during gestation. Functional observation battery: Forelimb grip strength in males during Week 7 showed a dose-related decrease. There were no other dose-related observations on functional effects.
Locomotor activity: No effects were observed. Organ weight effects observed in kidneys, adrenal glands and liver. Adjusted mean kidney weight was increased in high dose 350mg/kg males Adjusted mean adrenal weight was increased in high dose females( P<0.05). Adjusted mean liver weight was increased in all treated male groups at 70, 155 and 350mg/kg groups. Necropsy: No necropsy findings that were deemed to be treatment-related. There were treatment-related findings in the liver, kidney and lungs. Centrilobular hypertrophy was recorded in the majority of the high 350mg/kg dose male and female livers. In the kidney there was an increase in the level of hyaline droplets in males at 350mg/kg dose and In the lungs there was a minor increase in the level of foamy histiocytes in males and females from 350mg/kg dose group.
Number of abortions:
not specified
Pre- and post-implantation loss:
effects observed, treatment-related
Description (incidence and severity):
For the high 350mg/kg dose, numbers of implantations was lower than control animals
Total litter losses by resorption:
no effects observed
Early or late resorptions:
no effects observed
Dead fetuses:
effects observed, treatment-related
Description (incidence and severity):
Total embryo foetal loss occurred for one high 350mg/kg dose female.
Changes in pregnancy duration:
not specified
Changes in number of pregnant:
not specified
Other effects:
no effects observed
Description (incidence and severity):
No treatment-related effects on numbers of males and females mating or on male or female fertility indices.
Details on maternal toxic effects:
Testicular staging: Qualitative testis staging did not indicate any abnormalities in the integrity of the various cell types present within the different stages of the spermatogenic cycle.
Dose descriptor:
NOAEL
Effect level:
350 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
other: no advser effect observed
Abnormalities:
not specified
Fetal body weight changes:
effects observed, treatment-related
Description (incidence and severity):
For the 350mg/kg dose group, Day 4 weights were lower than controls
Reduction in number of live offspring:
effects observed, treatment-related
Description (incidence and severity):
Total embryo foetal loss occurred for one high 350mg/kg dose female.
Changes in sex ratio:
not specified
Changes in litter size and weights:
effects observed, treatment-related
Description (incidence and severity):
litter sizes was lower in 350mg/kg group than controls. .
Total litter loss occurred for two females in the high 350mg/kg dose group and one from each of the 70 and 155mg/kg and dose groups
Changes in postnatal survival:
effects observed, treatment-related
Description (incidence and severity):
A reduction in the pups viability in the high dose 350mg/kg group was observed when compared to control animals.
External malformations:
no effects observed
Description (incidence and severity):
Malformed/ shortened limbs occurred in one pup in the 155mg/kg and one pup in the 350mg/kg groups, however as this effect occurred in only one pup in one litter in these groups, the toxicological significance is not clear.
Skeletal malformations:
not specified
Visceral malformations:
not specified
Other effects:
effects observed, treatment-related
Description (incidence and severity):
For the high 350mg/kg dose, numbers of implantations was lower than control animals
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:yes
Details on embryotoxic / teratogenic effects:
At the 350mg/kg dose tested, the test material reduces pup viability after delivery
Dose descriptor:
NOAEL
Effect level:
155 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
other: teratogenicity
Dose descriptor:
LOAEL
Effect level:
350 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
other: teratogenicity
Abnormalities:
not specified
Developmental effects observed:
not specified
Treatment related:
not specified
Conclusions:
The NOAEL and LOAEL for rats was considered to be 155 mg/kg/day and 350mg/kg/day respectively.
Executive summary:

In the present study, the effect of test chemical on repeated administrationof the test chemical and reproductive developmental toxicity study performed by oral administration. The test chemical was administered to rats at doses 0, 70, 155, 350 mg/kg bw/day for approximately 8 weeks.The animals observed for clinical signs of toxicity; mortality; feed consumption; change in body weight; behaviour such as locomotor activity and grip strength; gross necropsy examination histopathological examination and reproductive performance were examined in the present study. The results of study revealed, no mortality during the period of study and one 350mg/kg female, on days 4 and 5 of dosing, showed post-dosing signs of subdued behaviour, twitching, semi-closed eyes, piloerection, was cold and showed staggered movement.The body weight results revealed majority of males in 350mg/kg group lost weight during the first week of dosing. Although mean body weights increased every week after that, the high dose males were approximately 9% lighter than control males by the end of the study. Group mean body weight gain was lower in high 350mg/kg dose females than controls during gestation. 155 and 350mg/kg dose females had lower mean body weight gain than controls during lactation. Feed consumption was observed to be reduced in 350mg/kg group animals as compared to control animals. Forelimb grip strength in males during Week 7 but there were no other dose-related observations on functional effects including locomotor activity at dose levels of 70, 155 and 350mg/kg. Organ weight change were seen in kidney, liver and adrenal weight, significantly increase were observed in high dose 350mg/kg group animals as compared to control animals.   No gross necropsy findings were seen that attributed to be treatment-related at any dose group at 70, 155 and 350mg/kg.The reproductive findings of the study showed no indication of any qualitative abnormality in the various cell types present within the different stages of the spermatogenic cycle and No treatment-related effects on numbers of males and females mating or on male or female fertility indices. The F1 generation results from parent rats revealed reduction in the pups viability in the 350mg/kg dose group. The litter data shown numbers of implantations, litter sizes and Day 4 weights were all lower than controls in the high dose 350mg/kg. Total litter loss occurred for two females in the 350mg/kg and one from each of the 70 and 155mg/kg dose groups.Total embryo foetal loss occurred for one female from 350mg/kg group. Malformed/ shortened limbs occurred in one pup in the 155mg/kg and one pup in the 350mg/kg dose group, however as this effect occurred in only one pup in one litter in these groups, the toxicological significance is not clear. From the observations and result of the study, The NOAEL and LOAEL for rats was considered to be 155 mg/kg/day  and 350mg/kg/day respectively in rats.

Reason / purpose:
read-across source
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 a NTP report.
Principles of method if other than guideline:
Test chemical : a cumulative neurotoxicant was evaluated for developmental toxicity in timed-pregnant Sprague-Dawley (CD) rats. Test chemical was dissolved in distilled water to yield doses of 0. 2.5, 7.5. and 15 mg/kg and administered once daily by gavage on gestational days (gd) 6 through 20.
GLP compliance:
not specified
Limit test:
no
Specific details on test material used for the study:
- Molecular weight (if other than submission substance): 71.08 g/mol
- Substance type: organic
- Physical state: solid
-Purity: 95%
Species:
rat
Strain:
Sprague-Dawley
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: the Charles River Laboratories. Inc.
- Age at study initiation: 8-12 weeks
- Weight at study initiation: 201 g to 263 g
- Housing: Males were housed singly in solid bottom polycarbonate cages (19" x 10 1/2" x 8") and females were group housed (maximum, 3 per cage) in solid bottom polycarbonate cages (19" x 10 1/2" x 8") with stainless steel wire lids (Laboratory Products. Rochelle Park. NJ).
- Diet (e.g. ad libitum):Purin Certified Rodent Chow, ad libitum
- Water (e.g. ad libitum): deionized/filtered water, ad libitum
- Acclimation period: No data available

ENVIRONMENTAL CONDITIONS
- Temperature (°C): ranged between 67.7 and 76.8 deg F
- Humidity (%): maintained at an average of 49.0%
- Air changes (per hr): Air in each animal room was exchanged at least 12 to 14 times per hour.
- Photoperiod (hrs dark / hrs light): 12-hrs light/12-hrs dark
Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
VEHICLE
- Justification for use and choice of vehicle (if other than water): Distilled water
- Concentration in vehicle: 0, 2.5. 7.5 or 15 mg/kg/day
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Prior to initiation of dosing, an aliquot of the control and each test chemical formulation was analyzed for the test chemical. Formulations that assayed at 90-110% of the nominal concentration were considered acceptable for use. All other formulations were remixed and reanalyzed prior to administration. Additional samples of each formulation were analyzed after use at the end of the dosing period to confirm stability. Duplicate aliquots of the predosing and postdosing samples were refrigerated and stored as archival samples.
Details on mating procedure:
- M/F ratio per cage: 1:1
- Length of cohabitation: overnight
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy : Female were examined by vaginal smear for the presence of sperm.
- After successful mating each pregnant female was caged (how): Sperm-positive females were individually housed until scheduled sacrifice on gestation day 20.
Duration of treatment / exposure:
Gestation day 6 through 20
Frequency of treatment:
Once daily
Duration of test:
16 days
Remarks:
Doses / Concentrations:
0, 2.5. 7.5 and 15 mg/kg/day
Basis:
nominal in water
No. of animals per sex per dose:
Control: 30 females
2.5 mg/kg/day: 29 females
5 mg/kg/day: 30 females
15 mg/kg/day: 29 females
Control animals:
yes, concurrent vehicle
Details on study design:
The oral route of administration corresponds to one of the potential routes of exposure to ACRL in the human population. Gavage was selected as the method of oral administration to allow accurate control of the dose administered. The doses selected for the dose range finding study were 0, 2.5, 5.0, 15, 30 and 60 mg/kg/day. The upper end of the proposed dosage range was expected to equal or exceed the rat LD10 based on rat mortality data from the literature. The lower end of the range included doses predicted to have no toxic effects on maternal or developmental parameters.
Maternal examinations:
Body weight for pregnant female was recorded on the mornings of gestation day 0 and 6-20, and immediately following sacrifice on gestation day 20. Spermpositive females were observed for clinical signs at and 4 hours after dosing from gestation day 6 through gestation day 19. Before dosing on the morning of gestation day 20, clinical signs were recorded. Animals that died between gestation day 0 and scheduled sacrifice on gestation day 20 were necropsied in order to determine the cause of death.

Uteri which presented no visible implantation sites were stained with ammonium sulfide (10%) in order to visualize any implantation sites which may have undergone very early resorption. Total number of corpora lutea, liver weight, gravid uterine weight and uterine contents were examined.
Ovaries and uterine content:
Uteri which presented no visible implantation sites were stained with ammonium sulfide (10%) in order to visualize any implantation sites which may have undergone very early resorption. Total number of corpora lutea, liver weight, gravid uterine weight and uterine contents were examined.
Fetal examinations:
Live fetuses were dissected from the uterus and immediately placed on a moist paper towel over a tray of ice to induce terminal cold anesthesia. Following terminal anesthesia and gross examination, the fetuses were sexed, weighed and dissected for visceral examination.
Statistics:
Parametric statistical procedures were applied to selected measures from the teratology study
Indices:
No data
Historical control data:
No data
Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality:
not specified
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Maternal toxicity during the test chemicalexposure was expressed primarily by decreased weight gain at the high dose. Maternal weight gain during treatment and gestation as well as weight gain corrected for gravid uterine weight exhibited decreasing trends. Maternal weight gain during treatment was reduced below control values in the high dose group, and corrected weight gain was reduced in the 7.5 and 15 mg/kg dose groups
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
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
Gravid uterine and liver weights were not measurably affected by the test chemical treatment.
Gross pathological findings:
not specified
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Details on results:
No Data Available
Number of abortions:
effects observed, non-treatment-related
Description (incidence and severity):
The percentages of resorptions. dead fetuses or malformed fetuses per litter, as well as fetal body weights, were statistically similar among the dose groups.
Pre- and post-implantation loss:
not specified
Total litter losses by resorption:
effects observed, non-treatment-related
Description (incidence and severity):
The percentages of resorptions. dead fetuses or malformed fetuses per litter, as well as fetal body weights, were statistically similar among the dose groups.
Early or late resorptions:
effects observed, non-treatment-related
Description (incidence and severity):
The percentages of resorptions. dead fetuses or malformed fetuses per litter, as well as fetal body weights, were statistically similar among the dose groups.
Dead fetuses:
effects observed, non-treatment-related
Description (incidence and severity):
The percentages of resorptions. dead fetuses or malformed fetuses per litter, as well as fetal body weights, were statistically similar among the dose groups.
Changes in pregnancy duration:
not specified
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:
Maternal toxicity during ACRL exposure was expressed primarily by decreased weight gain at the high dose. Maternal weight gain during treatment and gestation as well as weight gain corrected for gravid uterine weight exhibited decreasing trends. Maternal weight gain during treatment was reduced below control values in the high dose group, and corrected weight gain was reduced in the 7.5 and 15 mg/kg dose groups. Gravid uterine and liver weights were not measurably affected by ACRL treatment. In addition, other clinical signs were observed infrequently and exhibited no clear-cut relationship to ACRL dose.
Dose descriptor:
NOAEL
Effect level:
2.5 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
other: maternal toxicity
Remarks on result:
other: Not Specified
Abnormalities:
not specified
Fetal body weight changes:
not specified
Reduction in number of live offspring:
no effects observed
Changes in sex ratio:
not specified
Changes in litter size and weights:
not specified
Changes in postnatal survival:
not specified
External malformations:
no effects observed
Skeletal malformations:
no effects observed
Visceral malformations:
no effects observed
Other effects:
not specified
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects
Details on embryotoxic / teratogenic effects:
The test chemical treatment during gestation did not alter measured endpoints of embryo/fetal viability, growth or development. The percentages of resorptions. dead fetuses or malformed fetuses per litter, as well as fetal body weights, were statistically similar among the dose groups. ACRL treatment had no effect on the incidence of skeletal. visceral or external malformations. There was a dose-related trend toward an increased incidence of variations, but there were no statistically significant pairwise comparisons.
Dose descriptor:
NOAEL
Effect level:
15 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: developmental toxicity
Abnormalities:
not specified
Developmental effects observed:
not specified
Treatment related:
not specified
Conclusions:
The No observed effect level (NOAEL) for the test chemical in rats was observed to be 15 mg/kg/day for developmental toxicity and 2.5 mg/kg/day for maternal toxicity.
Executive summary:

The test chemical, a cumulative neurotoxicant was evaluated in a 1 generation study for developmental toxicity in pregnant Sprague-Dawley (Crl:CD BR VAF/Plus) rats. The test chemical was administered once daily by gavage on gestation day 6 through gestation day 20 in the concentration of 0, 2.5, 7.5 and 15 mg/kg. Maternal weight gain during treatment and gestation, as well as weight gain corrected for gravid uterine weight, exhibited decreasing trends. Maternal weight gain during treatment was reduced below control values in the high dose group, and corrected weight gain was reduced in the 7.5 and 15 mg/kg dose groups. The test chemical treatment during gestation did not alter measured endpoints of embryo/fetal viability, growth or development. The percentages of resorptions, dead fetuses or malformed fetuses per litter, as well as fetal body weights, were statistically similar among the dose groups. The test chemical treatment had no effect on the incidence of skeletal, visceral or external malformations. There was a dose-related trend toward an increased incidence of variations, but there were no statistically significant pairwise comparisons through 20.The No observed effect level (NOAEL) for the test chemical in rats was observed to be15 mg/kg/day. The No observed effect level (NOAEL) for acrylamide in rats was observed to be 2.5 mg/kg/day for maternal toxicity.

Data source

Referenceopen allclose all

Reference Type:
publication
Title:
Unnamed
Year:
1992
Reference Type:
other: Secondary source
Title:
Oral (gavage) combined repeat dose toxicity and reproduction/development toxicity screening test of test chemical in the rats.
Author:
J Rhodes.
Year:
2004
Bibliographic source:
Covance Laboratories Inc (2004). - As cited in HPV program document
Reference Type:
publication
Title:
Unnamed
Year:
1988

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Principles of method if other than guideline:
The above experiments were performed to evaluate and assess the developmental toxicity of the test chemical in the test animals.
GLP compliance:
not specified

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
not specified
Details on test material:
Details on test material
- Name of test material (as cited in study report): tert-Butylacrylamide
- Molecular formula (if other than submission substance): C7-H13-N-O
- Molecular weight (if other than submission substance): 127.1857g/mol
- Substance type: Organic
- Physical state: No Data Available
- Impurities (identity and concentrations): No Data Available
Specific details on test material used for the study:
- Molecular weight (if other than submission substance): 127.1857g/mol
- Substance type: Organic
- Physical state: No Data Available
- Impurities (identity and concentrations): No Data Available

Test animals

Species:
other: Study 2: Mouse; Study 3: rat; study 4: rat
Strain:
other: Study 2: Crl:CD-le(ICR) BR VAF/Plus; Study 2: Crl:WI(Glx/BRL/Han)BR; Study 4: Sprague-Dawley
Details on test animals and environmental conditions:
Study 2: TEST ANIMALS
- Source: Charles River laboratories, Inc. (Raleigh, NC).
- Age at study initiation: Females (8-10 weeks); males (10-12 weeks
- Weight at study initiation: Weight Range for Females: 20-35 g on gestation day 0 (day of vaginal plug detection
- Fasting period before study: No data
- Housing: Solid bottom polycarbonate cages with stainless steel wire lids (Laboratory Products, Rochelle Park, NJ).
- Use of restrainers for preventing ingestion (if dermal):no data
- Diet (e.g. ad libitum): Purina ·Certified Rodent Chowe (15002) (Ralston Purina Co., St. Louis, Missouri) available ad libitum to males and females (pelleted chow)
- Water (e.g. ad libitum): deionized/filtered water provided ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 69-75 F
- Humidity (%): 30-70% relative humidity
- Air changes (per hr): 12-14 air changes per hour
- Photoperiod (hrs dark / hrs light):12/12hrs of light and dark

Study 3: - At study initiation the rats were 13 to 15 weeks old.

Study 4: TEST ANIMALS
- Source: the Charles River Laboratories. Inc.
- Age at study initiation: 8-12 weeks
- Weight at study initiation: 201 g to 263 g
- Housing: Males were housed singly in solid bottom polycarbonate cages (19" x 10 1/2" x 8") and females were group housed (maximum, 3 per cage) in solid bottom polycarbonate cages (19" x 10 1/2" x 8") with stainless steel wire lids (Laboratory Products. Rochelle Park. NJ).
- Diet (e.g. ad libitum):Purin Certified Rodent Chow, ad libitum
- Water (e.g. ad libitum): deionized/filtered water, ad libitum
- Acclimation period: No data available

ENVIRONMENTAL CONDITIONS
- Temperature (°C): ranged between 67.7 and 76.8 deg F
- Humidity (%): maintained at an average of 49.0%
- Air changes (per hr): Air in each animal room was exchanged at least 12 to 14 times per hour.
- Photoperiod (hrs dark / hrs light): 12-hrs light/12-hrs dark

Administration / exposure

Route of administration:
other: Study 2, 3 and 4: Oral; gavage
Vehicle:
other: study 2 and 4: Distliled water; study 3: not specified
Details on exposure:
Study 2: PREPARATION OF DOSING SOLUTIONS: Test chemical will be
dissolved in distilled water to a concentration determined by the following
formula:
concentration (mg/ml) = dose level (mg/kg)/ dose volume (10.0 ml/kg)
Test chemical will be weighed into a volumetric flask and distilled water will be added to slightly below the mark. test chemical will be dispersed into a visually uniform solution and the flask will then be filled to the mark and agitated a second time. The solution will be sonicated, if necessary, to dissolve the chemical. 'Each concentration of test chemical will be formulated independently once during each' study replicate in a quantity sufficient for use across the entire dosing period for each replicate.
- Storage condition of formulation: Dose formulations will be stored at refrigeratea temperatures in 60 ml amber vials

Study 3: Dose volume was 5mL/kg.

Study 4: VEHICLE
- Justification for use and choice of vehicle (if other than water): Distilled water
- Concentration in vehicle: 0, 2.5. 7.5 or 15 mg/kg/day
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Study 2: Prior to administration and after administration, each formulation were analyzed to ensure a concentration between 90 and 110% of the target concentration.

Study 3: Achieved concentrations were 90 to 100% of expected when measured on Week 1 and Week 8

Study 4: Prior to initiation of dosing, an aliquot of the control and each ACRL formulation was analyzed for the test chemical. Formulations that assayed at 90-110% of the nominal concentration were considered acceptable for use. All other formulations were remixed and reanalyzed prior to administration. Additional samples of each formulation were analyzed after use at the end of the dosing period to confirm stability. Duplicate aliquots of the predosing and postdosing samples were refrigerated and stored as archival samples.
Details on mating procedure:
Study 2: Monogamous breeding pairs were cohabited over night in the home cage of the male. On the following morning, the female was examined for a copulation plug. The day of plug detection was designated as gd 0. Plug-negative females was returned to the males cage and checked for plugs on successive mornings until insemina tion occurs or the treatment groups are filled, whichever comes first.

Study 3: No data
- Impregnation procedure: cohoused
- M/F ratio per cage: 1/1 of same treatment group
- Length of cohabitation: 15 days
- Proof of pregnancy: Mating was confirmed by the presence of a vaginal plug in situ or by sperm in a vaginal washing. The day on which mating was confirmed was designated Day 0 of gestation

Study 4: - M/F ratio per cage: 1:1
- Length of cohabitation: overnight
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy : Female were examined by vaginal smear for the presence of sperm.
- After successful mating each pregnant female was caged (how): Sperm-positive females were individually housed until scheduled sacrifice on gestation day 20.
Duration of treatment / exposure:
Study 2: 11 days (from gestation period day 6 to 17)

Study 3: Males - 2 weeks before pairing, throughout pairing and until the day before necropsy (at least 4 weeks of treatment)
Females- 2 weeks before pairing, throughout pairing and until Day 4 postpartum, inclusive

Study 4: Gestation day 6 through 20
Frequency of treatment:
Study 2, 3 and 4: daily
Duration of test:
Study 2: 17 days (examination of fetuses were performed on day 17)
Study 3: Approximately 8 weeks
Study 4: 16 days
Doses / concentrationsopen allclose all
Remarks:
Study 2: 0, 30, 60, 120, and 180 mg/kg/day
Remarks:
Study 3: Doses / Concentrations:
0, 70, 155, 350 mg/kg bw/day
Basis:
no data
Remarks:
Study 4: Doses / Concentrations:
0, 2.5. 7.5 and 15 mg/kg/day
Basis:
nominal in water
No. of animals per sex per dose:
Study 2: 20 pregnant mice in each group
Study 3: 10 animals/sex/dose
Study 4: Control: 30 females
2.5 mg/kg/day: 29 females
5 mg/kg/day: 30 females
15 mg/kg/day: 29 females
Control animals:
other: Study 2 and 4: yes, concurrent vehicle; study 3: yes, 5% v/v etahnol in corn oil
Details on study design:
Study 2: Administration by gavage on the mornings of gd 6 through 17, using an 18-gauge 1.5 inch stainless steel dosing tube, dose volume was 10ml/kg. On gestational day 17, approximately one to one and a half days before expected parturition. pregnant dams were anesthetized by carbon dioxide gas and sacrificed by cervical dislocation. The uterus and right ovary were removed and fetus examination were performed

Study 3: No data available

Study 4: The oral route of administration corresponds to one of the potential routes of exposure to ACRL in the human population. Gavage was selected as the method of oral administration to allow accurate control of the dose administered. The doses selected for the dose range finding study were 0, 2.5, 5.0, 15, 30 and 60 mg/kg/day. The upper end of the proposed dosage range was expected to equal or exceed the rat LD10 based on rat mortality data from the literature. The lower end of the range included doses predicted to have no toxic effects on maternal or developmental parameters.

Examinations

Maternal examinations:
Study 2: Feed comsumption, water consumption, Body weight, liver, and uterus of each female were recorded on the mornings of gd 0, 3, 6-17 and 1mmediately following sacrifice on gd 17.

Study 3: one 350mg/kg female, on days 4 and 5 of dosing, showed post-dosing signs of subdued behaviour, twitching, semi-closed eyes, piloerection, was cold and showed staggered movement.

Study 4: Body weight for pregnant female was recorded on the mornings of gestation day 0 and 6-20, and immediately following sacrifice on gestation day 20. Spermpositive females were observed for clinical signs at and 4 hours after dosing from gestation day 6 through gestation day 19. Before dosing on the morning of gestation day 20, clinical signs were recorded. Animals that died between gestation day 0 and scheduled sacrifice on gestation day 20 were necropsied in order to determine the cause of death.
Uteri which presented no visible implantation sites were stained with ammonium sulfide (10%) in order to visualize any implantation sites which may have undergone very early resorption. Total number of corpora lutea, liver weight, gravid uterine weight and uterine contents were examined.
Ovaries and uterine content:
Study 2: The uterus and right ovary were removed and uterine contents examined to determine the number of implantation sites, resorptions, dead fetuses, and live fetuses.

Study 4: Uteri which presented no visible implantation sites were stained with ammonium sulfide (10%) in order to visualize any implantation sites which may have undergone very early resorption. Total number of corpora lutea, liver weight, gravid uterine weight and uterine contents were examined.
Fetal examinations:
Study 2: fetal examination were performed including head, external and visceral examination.

Study 4: Live fetuses were dissected from the uterus and immediately placed on a moist paper towel over a tray of ice to induce terminal cold anesthesia. Following terminal anesthesia and gross examination, the fetuses were sexed, weighed and dissected for visceral examination.
Statistics:
Study 2: Arcsine, Square root transformation, Bartletts test, ANOVA and chi- square test were used for statistical analysis

Study 4: Parametric statistical procedures were applied to selected measures from the teratology study
Indices:
No Data Available
Historical control data:
No Data Available

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Study 2: Mouth bleeding before dosing, Cyst on right ovary,Left ovary enclosed in blood· filled cyst were observed at 30mg/kg group; Cyst on left ovary was observed in 60mg/kg group; Uterus enlarged, Rough coat,Vaginal bleeding,Cyst on right ovary were observed in 120mg/kg group females perioral wetness, Vaginal discharge (brownish red), Lethargic,Convulsing, Cyst on right ovary, Curling hind limbs under bodywere obseved in treated females but this effects were non significant compared to control female mice.
Dermal irritation (if dermal study):
not specified
Mortality:
no mortality observed
Description (incidence):
Study 2: No mortality were observed at 30, 60, 120 and 180mg/kg dose groups
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Study 2: No significant change in body weight were observed at 30, 60, 120 and 180mg/kg dose groups compared to control
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
Study 2: No significant change in feed consumption were observed at 30, 60, 120 and 180mg/kg dose groups compared to control
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
no effects observed
Description (incidence and severity):
Study 2: No significant change in water consumption were observed at 30, 60, 120 and 180mg/kg dose groups compared to control
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
effects observed, non-treatment-related
Description (incidence and severity):
Study 3: Functional observation battery: Forelimb grip strength in males during Week 7 showed a dose-related decrease. There were no other dose-related observations on functional effects.
Locomotor activity: No effects
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
Study 2: Significant change in liiver and gravid uterus were observed in 180mg/kg group animals

Study 4: Organ weight effects observed in kidneys, adrenal glands and liver. Adjusted mean kidney weight was increased in high dose 350mg/kg males Adjusted mean adrenal weight was increased in high dose females( P<0.05). Adjusted mean liver weight was increased in all treated male groups at 70, 155 and 350mg/kg groups
Gross pathological findings:
no effects observed
Description (incidence and severity):
Study 3: Necropsy: No necropsy findings that were deemed to be treatment-related.
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Study 3: Histopathology:There were treatment-related findings in the liver, kidney and lungs. Centrilobular hypertrophy was recorded in the majority of the high 350mg/kg dose male and female livers. In the kidney there was an increase in the level of hyaline droplets in males at 350mg/kg dose and In the lungs there was a minor increase in the level of foamy histiocytes in males and females from 350mg/kg dose group.
Histopathological findings: neoplastic:
not specified
Other effects:
not specified

Maternal developmental toxicity

Number of abortions:
not specified
Pre- and post-implantation loss:
not specified
Total litter losses by resorption:
not specified
Early or late resorptions:
not specified
Dead fetuses:
not specified
Changes in pregnancy duration:
not specified
Changes in number of pregnant:
not specified
Other effects:
no effects observed
Description (incidence and severity):
Study 3: Reproductive performance: No treatment-related effects on numbers of males and females mating or on male or female fertility indices

Details on maternal toxic effects:
Study 3: Testicular staging: Qualitative testis staging did not indicate any abnormalities in the integrity of the various cell types present within the different stages of the spermatogenic cycle.

Study 4: Maternal toxic effects:no effects
Details on maternal toxic effects:
Maternal toxicity during test chemical exposure was expressed primarily by decreased weight gain at the high dose. Maternal weight gain during treatment and gestation as well as weight gain corrected for gravid uterine weight exhibited decreasing trends. Maternal weight gain during treatment was reduced below control values in the high dose group, and corrected weight gain was reduced in the 7.5 and 15 mg/kg dose groups. Gravid uterine and liver weights were not measurably affected by test chemical treatment. In addition, other clinical signs were observed infrequently and exhibited no clear-cut relationship to test chemical dose.

Effect levels (maternal animals)

Dose descriptor:
NOAEL
Effect level:
180 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
behaviour (functional findings)
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic

Maternal abnormalities

Abnormalities:
not specified

Results (fetuses)

Fetal body weight changes:
effects observed, treatment-related
Description (incidence and severity):
Study 3: For the 350mg/kg dose group, Day 4 weights were lower than controls
Reduction in number of live offspring:
effects observed, treatment-related
Description (incidence and severity):
Study 3: Total embryo foetal loss occurred for one high 350mg/kg dose female.
Changes in sex ratio:
not specified
Changes in litter size and weights:
effects observed, treatment-related
Description (incidence and severity):
Study 3: litter sizes was lower in 350mg/kg group than controls. .
Total litter loss occurred for two females in the high 350mg/kg dose group and one from each of the 70 and 155mg/kg and dose groups
Changes in postnatal survival:
effects observed, treatment-related
Description (incidence and severity):
Study 3: a reduction in the pups viability in the high dose 350mg/kg group was observed when compared to control animals
External malformations:
effects observed, non-treatment-related
Description (incidence and severity):
Study 2: Cleft Palate, Ectocardia, Gastroschisis and Short Tail were obseved in 2 fetuses

Study 3: Malformed/ shortened limbs occurred in one pup in the 155mg/kg and one pup in the 350mg/kg groups, however as this effect occurred in only one pup in one litter in these groups, the toxicological significance is not clear.
Skeletal malformations:
effects observed, treatment-related
Description (incidence and severity):
Study 2: Asymmetrical Fusion of Sternal Plate, Cleft Sternum, Perforated Sternum, Fused Rib Cartilage, Incomplete Ossification, Cartilage Present, Extra Ossification Site(s), Hematoma: Head, Face, Jaw, Neck, Abdomen, Thorax, Hindlimb were observed.
Visceral malformations:
effects observed, treatment-related
Description (incidence and severity):
Study 2: Situs Inversus Viscerum and left hydronephrosis noted in treated animals
Other effects:
effects observed, treatment-related
Description (incidence and severity):
Study 3: For the high 350mg/kg dose, numbers of implantations was lower than control animals
Details on embryotoxic / teratogenic effects:
Study 3: Embryotoxic / teratogenic effects:yes
Details on embryotoxic / teratogenic effects:
At the 350mg/kg dose tested, the test material reduces pup viability after delivery

Study 4: Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
ACRL treatment during gestation did not alter measured endpoints of embryo/fetal viability, growth or development. The percentages of resorptions. dead fetuses or malformed fetuses per litter, as well as fetal body weights, were statistically similar among the dose groups. ACRL treatment had no effect on the incidence of skeletal. visceral or external malformations. There was a dose-related trend toward an increased incidence of variations, but there were no statistically significant pairwise comparisons.

Effect levels (fetuses)

Dose descriptor:
NOAEL
Effect level:
120 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: The results is based on the effects on teratoloical parameters.
Remarks on result:
other: Not Specified

Fetal abnormalities

Abnormalities:
not specified

Overall developmental toxicity

Developmental effects observed:
not specified
Treatment related:
not specified

Applicant's summary and conclusion

Conclusions:
Based on all the observations and results, it was observed that, the NOAEL for the test chemical for reproductive parameters of parental animals was found to be 180 mg/kg bw and NOAEL for the developmental parameters fetuses was found to be 120 mg/kg bw.
Executive summary:

Developmental Toxicity Study:

The data for developmental toxicity as summaries is given below:

Developmental Toxicity Study 2:

In the present study, developmental Toxicity of test chemical was evaluated by adinistration of test chemical by Gavage to Swiss CCD-I (trade name)) Mice on Gestational Days 6 through 17. The dose levels of 0, 30, 60, 120, 180mg/kg were selected and administered in pregnant female mice at day 6 to 17 of gestation period. Dams were observed for clinical signs, change in body weight, feed and water consumption and organ weight change specially liver and gravid uterus. The fetuses were observed for external, visceral and skeletal abnormalities.

The results of the study revealed clinical toxicity life, Cyst on right ovary,Left ovary enclosed in blood· filled cyst  were observed at 30mg/kg group; Cyst on left ovary was observed in 60mg/kg group; Uterus enlarged, Rough coat,Vaginal bleeding,Cyst on right ovary were observed in 120mg/kg group females perioral wetness, Vaginal discharge (brownish red), Lethargic,Convulsing, Cyst on right ovary, Curling hind limbs under bodywere obseved in treated females but this effects were non significant compared to control female mice but no change in body weight change, feed consumption and water consumption were observed maternal animals at 30,60,120 and 180mg/kg groups.

The fetuses observed for external , visceral and skeletal abnormalities, the results were Cleft Palate, Ectocardia, Gastroschisis and Short Tail were obseved in 2 fetuses. Skeletal malformations were observed such as asymmetrical Fusion of Sternal Plate, Cleft Sternum, Perforated Sternum, Fused Rib Cartilage, Incomplete Ossification, Cartilage Present, Extra Ossification Site(s), Hematoma: Head, Face, Jaw, Neck, Abdomen, Thorax, Hindlimb. the visceral abnormalities also obseved in treated animals including, Situs Inversus Viscerum and left hydronephrosis. From the observations and results of the study, The NOAEL value from the maternal observations was considered to be 180mg/kg/day howeveer the NOAEL value from the fetuses observation considered to be 120mg/kg.

Developmental Toxicity Study 3:

In the present study, the effect of test chemical on repeated administration of the test chemical and reproductive developmental toxicity study performed by oral administration. The test substance was administered to rats at doses 0, 70, 155, 350 mg/kg bw/day for approximately 8 weeks.The animals observed for clinical signs of toxicity; mortality; feed consumption; change in body weight; behaviour such as locomotor activity and grip strength; gross necropsy examination histopathological examination and reproductive performance were examined in the present study. The results of study revealed, no mortality during the period of study and one 350mg/kg female, on days 4 and 5 of dosing, showed post-dosing signs of subdued behaviour, twitching, semi-closed eyes, piloerection, was cold and showed staggered movement.The body weight results revealedmajority of males in 350mg/kg group lost weight during the first week of dosing. Although mean body weights increased every week after that, the high dose males were approximately 9% lighter than control males by the end of the study. Group mean body weight gain was lower in high 350mg/kg dose females than controls during gestation. 155 and 350mg/kg dose females had lower mean body weight gain than controls during lactation. Feed consumption was observed to be reduced in 350mg/kg group animals as compared to control animals. Forelimb grip strength in males during Week 7 but there were no other dose-related observations on functional effects including locomotor activity at dose levels of 70, 155 and 350mg/kg. Organ weight change were seen in kidney, liver and adrenal weight, significantly increase were observed in high dose 350mg/kg group animals as compared to control animals. No gross necropsy findings were seen that attributed to be treatment-related at any dose group at 70, 155 and 350mg/kg.The reproductive findings of the study showed no indication of any qualitative abnormality in the various cell types present within the different stages of the spermatogenic cycle and No treatment-related effects on numbers of males and females mating or on male or female fertility indices. The F1 generation results from parent rats revealed reduction in the pups viability in the 350mg/kg dose group. The litter data shown numbers of implantations, litter sizes and Day 4 weights were all lower than controls in the high dose 350mg/kg. Total litter loss occurred for two females in the 350mg/kg and one from each of the 70 and 155mg/kg dose groups.Total embryo foetal loss occurred for one female from 350mg/kg group. Malformed/ shortened limbs occurred in one pup in the 155mg/kg and one pup in the 350mg/kg dose group, however as this effect occurred in only one pup in one litter in these groups, the toxicological significance is not clear. From the observations and result of the study, The NOAEL and LOAEL for rats was considered to be 155 mg/kg/day  and 350mg/kg/day respectively in rats.

Developmental Toxicity Study 4:

The test chemical, a cumulative neurotoxicant was evaluated in a 1 generation study for developmental toxicity in pregnant Sprague-Dawley (Crl:CD BR VAF/Plus) rats. Acrylamide was administered once daily by gavage on gestation day 6 through gestation day 20 in the concentration of 0, 2.5, 7.5 and 15 mg/kg. Maternal weight gain during treatment and gestation, as well as weight gain corrected for gravid uterine weight, exhibited decreasing trends. Maternal weight gain during treatment was reduced below control values in the high dose group, and corrected weight gain was reduced in the 7.5 and 15 mg/kg dose groups. The test chemical treatment during gestation did not alter measured endpoints of embryo/fetal viability, growth or development. The percentages of resorptions, dead fetuses or malformed fetuses per litter, as well as fetal body weights, were statistically similar among the dose groups. The test chemical treatment had no effect on the incidence of skeletal, visceral or external malformations. There was a dose-related trend toward an increased incidence of variations, but there were no statistically significant pairwise comparisons. through20.The No observed effect level (NOAEL) for the test chemical in rats was observed to be15 mg/kg/day. The No observed effect level (NOAEL) for acrylamide in rats was observed to be 2.5 mg/kg/day for maternal toxicity.