Registration Dossier

Toxicological information

Toxicity to reproduction

Currently viewing:

Administrative data

Endpoint:
screening for reproductive / developmental toxicity
Remarks:
based on generations indicated in Effect levels (migrated information)
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Cross-referenceopen allclose all
Reason / purpose for cross-reference:
reference to same study
Reason / purpose for cross-reference:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report date:
2010

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: Liquid
Details on test material:
Name of test material (as cited in study report): 4,4'-isopropylydenediphenol ethoxylated
Physical state: liquid
Analytical purity: >99%

Test animals

Species:
rat
Strain:
Crj: CD(SD)
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS

Source: Charles River Japan
Age at study initiation: 10 weeks
Weight at study initiation: Male 361 (334-396) g, Female 235 (216-260) g
Housing: 1/cage
Diet (e.g. ad libitum): MR(pellet), Nosan Corporation, Japan, ad libitum
Water (e.g. ad libitum): tap water(filtered and UV sterilized), ad libitum
Acclimation period: 12 days

ENVIRONMENTAL CONDITIONS

Temperature (°C): 22.1-25.0
Humidity (%): 46-60
Air changes (per hr): >10/hr
Photoperiod (hrs dark / hrs light): 12 hr/12 hr

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
olive oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:

VEHICLE

Concentration in vehicle: 0, 30, 120, 500, 1000 mg/kg/day
Amount of vehicle (if gavage): 5 mL/kg
Lot/batch No. (if required): BH17
Analytical verification of doses or concentrations:
yes
Duration of treatment / exposure:
Males: 42 days
Females: 42-47 days, from 14 days before mating to day 4 of lactation
Frequency of treatment:
Once a day
Details on study schedule:
12 of each sex exposed to 0, 30, 120, 500 and 1000 mg/kg bw per day.
Doses / concentrationsopen allclose all
Dose / conc.:
0 mg/kg bw/day (actual dose received)
Dose / conc.:
30 mg/kg bw/day (actual dose received)
Dose / conc.:
120 mg/kg bw/day (actual dose received)
Dose / conc.:
500 mg/kg bw/day (actual dose received)
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
12 male and 12 female per group, with additional 5 per sex at 0 and 1000 mg/kg recovery group.
Control animals:
yes, concurrent vehicle
Details on study design:
Satellite group doses: 0, 1000 mg/kg
Post-exposure recovery period in satellite group: 14 days
Positive control:
No

Examinations

Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes

Time schedule:

Cage side observations checked in table were included.

DETAILED CLINICAL OBSERVATIONS: Yes

Time schedule: Day 1 of administration and once a week thereafter.



Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Slight salivation was observed at 500 mg/kg bw and 1,000 mg/kg bw in female and male rats during the administration period. No other significant clinical signs were detected in any of the test groups.
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Description (incidence):
There was no mortality recorded in either the male or female test groups.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
All animals gained body weight during the testing period, with the lowest body weight gain generally seen in the highest dose groups. The weight gain in the male group treated at 1000 mg/kg/bw was significantly (p<0.05) lower than the control group during the administration period. During the recovery period, animals from the 1,000 mg/kg bw had a significantly (p<0.05) higher body weight gain when compared to the control animals. Their final body weight was similar at the end of the recovery period between treated and control female and male rats.
Food consumption and compound intake (if feeding study):
effects observed, non-treatment-related
Description (incidence and severity):
On Day 35 of the administration period, food consumption of male rats treated at 500 mg/kg bw/d was significantly (p<0.05) lower than for the control animals. This difference was not significant on Day 41 at the same dose level, and on Days 35 and 41 at 1,000 mg/kg bw/d.
On Day 0 of lactation, food consumption of female rats treated at 500 mg/kg bw/d was significantly (p<0.01) higher than for the control animals. This difference was not significant on Day 3 of lactation at the same dose level, and on Days 0 and 3 at 1,000 mg/kg bw/d.
Food consumption of the female rats from the satellite group treated at 1,000 mg/kg bw/d was significantly (p<0.05) higher than for the control animals on Day 28 of administration. On Day 35 the food consumption was similar between the treated and control animals.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
no effects observed
Description (incidence and severity):
Eye sight reaction and pupil reflex recorded as normal in all groups. No eye discharge recorded.
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
At the end of the recovery period, the number of platelets was significantly (p<0.05) lower in male rats treated at 1,000 mg/kg bw when compared to the control animals. This effects was not observed at the end of the administration period.
At the end of the administration period, the red blood cytes count, hemoglobin, and hematocrit were significantly (respectively p<0.05, p<0.01, and p<0.05) lower to the control animals, while the percentage of reticulocytes was significantly (p<0.05) higher. At the end of the recovery period these altered parameters were comparable between the treated and control female rats, while the activated partial thromboplastin time was significantly (p<0.05) in the treated female rats.
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
At the end of the administration period, a significantly (p<0.05) lower creatinine level was observed in male rats treated at 30 mg/kg bw/d.
At the end of the administration period, a significantly (p<0.01) lower creatinine level was observed in male rats treated at 120 mg/kg bw/d.
At the end of the administration period, a significantly (p<0.01) lower creatinine level and significantly (p<0.05) higher total-cholesterol and calcium levels were observed in male rats treated at 500 mg/kg bw/d.
At the end of the administration period, a significantly (p<0.01) lower creatinine level and significantly (p<0.01) higher total-cholesterol level were observed in male rats treated at 1,000 mg/kg bw/d.
At the end of the recovery period, these altered parameters were comparable between the treated and control male rats, while the glucose level was significantly (p<0.05) lower and the alanine aminotrans ferase significantly (p<0.05) higher in the treated animals.

At the end of the administration period, levels of alanine aminotransferase, total cholesterol, and calcium were significantly (respectively p<0.05, p<0.01, p<0.01) higher in female rats treated at 1,000 mg/kg bw/d when compared to the control animals.
At the end of the recovery period, these altered parameters were comparable between the treated and control male rat, while the glucose level was significantly (p<0.05) lower and the alanine aminotransferase significantly (p<0.05) higher in the treated animals. At the end of the recovery period, these altered parameters were comparable between the treated and control female rats.
Urinalysis findings:
no effects observed
Behaviour (functional findings):
effects observed, treatment-related
Description (incidence and severity):
At the end of the recovery period, grip strength of hindlimbs and motor activity (30 minutes) were significantly (respectively p<0.01 and p<0.05) reduced in animal treated at 1,000 mg/kg bw/d when compared to the control animals. Grip strength of forelimbs and motor activity (60 min) were not significantly different between the treated and control animals.
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Hypertrophy, hepatocyte, centrilobular were observed in liver of female rats treated at 1,000 mg/kg bw/d. These were not apparent in recovery group animals suggesting transitory effects.
Slight to moderate changes were observed in kidneys of male rats treated at 1,000 mg/kg bw/d. These were not apparent in recovery group animals suggesting transitory effects.

No abnormalities were detected in the brain, thyroid, parathyroid, thymus, trachea, small intestine, large intestine, adrenal, urinary bladder, testis, epididymis, seminal vesicle, spinal cord, sciatic nerve, bone, bone marrow, lymph nodes and mammary gland from animals of control and 1000 mg/kg/bw groups of male rats. No abnormalities were detected in the brain, pituitary, thyroid, parathyroid, trachea, small intestine, large intestine, heart, adrenal, urinary bladder, ovary, uterus, spinal cord, sciatic nerve, bone, bone marrow, lymph nodes and mammary glands of control and 1000 mg/kg/bw groups of female rats.
Histopathological findings: neoplastic:
no effects observed
Other effects:
no effects observed

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
no effects observed
Description (incidence and severity):
The oestrus cycle was comparable between the treated and control female rats along with fertility index, number of corpora lutea and implantation sites, and gestation index.
Reproductive function: sperm measures:
no effects observed
Description (incidence and severity):
The number of cells in seminiferous epithelia was comparable between the male rats treated at 1,000 mg/kg bw/d and the control male rats. Animals treated at other concentrations were not investigated
Reproductive performance:
no effects observed
Description (incidence and severity):
The pairing until copulation and copulation index were comparable between the treated and control animals.

Effect levels (P0)

open allclose all
Key result
Dose descriptor:
NOAEL
Effect level:
> 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
reproductive function (oestrous cycle)
reproductive function (sperm measures)
reproductive performance
Dose descriptor:
NOAEL
Effect level:
500 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain

Target system / organ toxicity (P0)

open allclose all
Critical effects observed:
no
Lowest effective dose / conc.:
1 000 mg/kg bw/day (actual dose received)
System:
hepatobiliary
Organ:
liver
Critical effects observed:
no
Lowest effective dose / conc.:
1 000 mg/kg bw/day (actual dose received)
System:
urinary
Organ:
kidney

Results: F1 generation

General toxicity (F1)

Clinical signs:
no effects observed
Description (incidence and severity):
Sex ratio was comparable between the treated and control animals.
Dermal irritation (if dermal study):
not examined
Mortality / viability:
no mortality observed
Description (incidence and severity):
The number of pups born, live birth index, and viability index were comparable between the treated and control animals.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
The bodyweight of pups was comparable between the treated and control animals.
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Sexual maturation:
no effects observed
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
1/175 of the examined pups born from animals treated at 30 mg/kg bw/d showed an external malformation (missing tail). Pups from other groups including control did not display any observable external malformation. This malformation was not considered as significant and treatment-related.
Histopathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
1/120 of the examined pups born from animals treated at 120 mg/kg bw/d and 2/146 of the examined pups born from animals treated at 500 mg/kg bw/d showed a visceral malformation (persistent left umbilical artery). Pups from other groups including control did not display any observable external malformation. These malformations were not considered as significant and treatment-related.

Developmental neurotoxicity (F1)

Behaviour (functional findings):
not examined

Developmental immunotoxicity (F1)

Developmental immunotoxicity:
not examined

Effect levels (F1)

Key result
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
> 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
viability
clinical signs
mortality
body weight and weight gain
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic

Overall reproductive toxicity

Reproductive effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
The NOAEL for general toxicity on the parent animals was determined to be 500 mg/kg bw/d on the basis of the significant bodyweight reduction observed at 1,000 mg/kg bw/d. In regard to the toxicity to reproduction, no significant treatment-related effects were observed in parent animals and pups at concentrations up to 1,000 mg/kg bw/d, it is therefore proposed a NOAEL of 1,000 mg/kg bw/d for the toxicity to reproduction.
Executive summary:

A combined short-term toxicity and screening for toxicity to reproduction study was performed on BPA-5EO according to OECD Testing Guideline 422. Groups of 12 rats per sex were exposed for 42 days to the substance by the oral route. The concentrations selected for this study were 0, 30, 120, 500, and 1,000 mg/kg bw/d. Two additional groups of five animals per sex were included for a 14-day recovery period and received 0 or 1,000 mg/kg bw/d. Mortality, clinical signs, bodyweights, and food consumption were recorded during the study. Before termination samples of blood and urine were collected to investigate the haematology and blood chemistry parameters and perform the urinalysis. Ophthalmological examination and FOB were performed. At termination, pathological and histopathological examinations were undertaken and organs were weight.

No animal died as a result of the exposure to the substance at up to 1,000 mg/kg bw/d. Slight salivation was observed at 500 and 1,000 mg/kg bw but this was not considered as a significant effect and salivation stopped during the recovery period. A significantly reduced bodyweight gain was observed in male animals treated at 1,000 mg/kg bw/d however the bodyweight was comparable at the end of the recovery period between treated and control animals. A momentary reduced found consumption was observed at 500 mg/kg bw or 1,000 mg/kg bw during the administration period. Some changes were observed at 1,000 mg/kg bw/d during the FOB. Some parameters in the haematology, biochemistry, and urinalysis were altered at the end of the administration period but recovered. Reversible variations in organ weights were observed at the end of the administration period. No effects were observed during the ophthalmological examination. Changes were observed in liver of female rats treated at 1,000 mg/kg bw/d. Slight to moderate changes were observed in kidneys of male rats treated at 1,000 mg/kg bw/d. These were not apparent in recovery group animals suggesting transitory effects. No abnormalities were detected in the brain, thyroid, parathyroid, thymus, trachea, small intestine, large intestine, adrenal, urinary bladder, testis, epididymis, seminal vesicle, spinal cord, sciatic nerve, bone, bone marrow, lymph nodes and mammary gland from animals of control and 1000 mg/kg/bw groups of male rats. No abnormalities were detected in the brain, pituitary, thyroid, parathyroid, trachea, small intestine, large intestine. heart, adrenal, urinary bladder, ovary, uterus, spinal cord, sciatic nerve, bone, bone marrow, lymph nodes and mammary glands of control and 1000 mg/kg/bw groups of female rats.

The reproductive function and performance were comparable between the treated and control parent animals. Pups born from the different groups were in comparable number and did not display significant treatment-related mortality, clinical signs, effects on sex ratio or bodyweight, or malformations. 1/175 of the examined pups born from animals treated at 30 mg/kg bw/d showed an external malformation (missing tail). 1/120 of the examined pups born from animals treated at 120 mg/kg bw/d and 2/146 of the examined pups born from animals treated at 500 mg/kg bw/d showed a visceral malformation (persistent left umbilical artery). Pups from other groups including control did not display any observable external malformation. These malformations were not considered as significant and treatment-related.

The NOAEL for general toxicity on the parent animals was determined to be 500 mg/kg bw/d on the basis of the significant bodyweight reduction observed at 1,000 mg/kg bw/d. In regard to the toxicity to reproduction, no significant treatment-related effects were observed in parent animals and pups at concentrations up to 1,000 mg/kg bw/d, it is therefore proposed a NOAEL of 1,000 mg/kg bw/d for the toxicity to reproduction.