Registration Dossier

Diss Factsheets

Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information
 In the screening study on reproduction, Ethoxylated bisphenol A diacrylate was administered daily by oral gavage to male and female Sprague Dawley rats, for 2 weeks before pairing, during pairing, gestation and until day 5 p.p., at dose-levels of 50, 250 or 1000 mg/kg/day.
At 1000 mg/kg/day, round back and piloerection noted in both sexes in weeks 2/3, as well as the higher mean cholesterol level at the end of the treatment period were considered to be adverse. There were no adverse findings at 50 and 250 mg/kg/day. Therefore, based on the experimental conditions of this study: the NOAEL for parental toxicity was 250 mg/kg/day, the NOAEL for reproductive performance (mating and fertility) and the NOAEL for toxic effects on progeny were 1000 mg/kg/day.
Link to relevant study records
Reference
Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
24 September 2012 - 08 February 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
Deviations:
yes
Remarks:
addition of haematology and blood biochemistry analysis
GLP compliance:
yes (incl. QA statement)
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: breeder: Charles River Laboratories Italia, Calco, Italy
- Age at study initiation: on the first day of treatment, the males were approximately 10 weeks old and the females were approximately 9 weeks old
- Mean body weight at study initiation: on the first day of treatment, the males had a mean body weight of 373 g (range: 327 g to 401 g) and the females had a mean body weight of 214 g (range: 176 g to 248 g)
- Fasting period before study: no
- Housing: the females were individually housed, except during pairing and lactation, in polycarbonate cages. The males were individually housed except during pairing, in wire-mesh cages
- Diet: SSNIFF R/M-H pelleted diet (free access)
- Water: tap water filtered with a 0.22 µm filter (free access)
- Acclimation period: 7 days before the beginning of the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C
- Humidity (%): 50 ± 20%
- Air changes (per hr): approximately 12 cycles/hour of filtered, non-recycled air
- Photoperiod (hrs dark / hrs light): 12 h/12 h.

IN-LIFE DATES: 02 October 2012 to 02 December 2012.
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
PREPARATION OF DOSING FORMULATIONS:
The test item was administered as a suspension in the vehicle. It was mixed with the required quantity of vehicle. No correction factor was applied.
The frequency of dose formulation preparation was on a daily basis. The dose formulations were delivered to the study room at room temperature and protected from light.

VEHICLE
- Justification for use and choice of vehicle: homogenous suspension with test item and corn oil which is commonly used in this type of study
- Concentration in vehicle: 10, 50 and 200 mg/mL
- Amount of vehicle (if gavage): 5 mL/kg/day.
Details on mating procedure:
- M/F ratio per cage: 1/1
- Length of cohabitation (mating period): until mating occurred or 14 days has elapsed in the latter case, then pairing with a second male until mating occurred within 7 days
- Proof of pregnancy: vaginal plug or sperm in the morning vaginal lavage referred to as day 0 post-coitum
- After successful mating each pregnant female was caged individually
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Type of method: HPLC-UV
Test item concentrations: remained within an acceptable range of variation compared to nominal values.
Homogeneity: homogenous
Stability: not assessed, dose formulation prepared daily
Duration of treatment / exposure:
In the males:
- 2 weeks before mating,
- during the mating period,
- until sacrifice (at least 5 weeks in total),

In the females:
- 2 weeks before mating,
- during the mating period,
- during pregnancy,
- during lactation until day 5 post-partum inclusive,
- until sacrifice for females which had not delivered.
Frequency of treatment:
Daily
Details on study schedule:
- No F1 parents (only one generation mated)
- Age at mating of the mated animals in the study: 11-12 weeks
Dose / conc.:
50 mg/kg bw/day (actual dose received)
Dose / conc.:
250 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:
10 animals per sex per dose
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale:
The dose-levels were selected by the Sponsor, following the results of a previous 4 week toxicity study performed in the same species and strain. In this study, three groups of five males and five females received the test item by gavage at 100, 300 or 1000 mg/kg/day as a suspension in corn oil. Another group of five males and five females received the vehicle only. There were no premature deaths and the only test item-related clinical sign was ptyalism at 1000 mg/kg/day. There were no toxicologically significant effects on mean body weight and food consumption. Some slight effects were seen at hematology and biochemistry investigations, such as lower mean alkaline phosphatase activity in females treated at 1000 mg/kg/day, dose related increase in cholesterol level from 300 mg/kg/day or minimally higher mean red blood cell parameters in males treated at 1000 mg/kg/day. At necropsy, there was a higher mean liver weight particularly at 1000 mg/kg/day in males and females.

- Rationale for animal assignment: computerized stratification procedure.
Positive control:
no (not required).
Parental animals: Observations and examinations:
MORTALITY/MORBIDITY:
- Time schedule: at least twice a day during the treatment period.

CLINICAL OBSERVATIONS:
- Time schedule: once a day during the treatment period.

BODY WEIGHT:
- Time schedule: Males: on the first day of treatment, then once a week until sacrifice. Females: on the first day of treatment, then once a week until mating, on Days 0, 7, 14 and 20 post-coitum and days 1 and 5 post-partum.

FOOD CONSUMPTION:
- Time schedule: Males: on the first day of treatment, then once a week until pairing. Females: on the first day of treatment, then once a week until pairing, on Days 0, 7, 14 and 20 post-coitum and days 1 and 5 post-partum.

HAEMATOLOGY, CLINICAL CHEMISTRY:
- Time schedule: at the end of the treatment period.

REPRODUCTION (apart from indices):
- Pre-coital time and duration of gestation were recorded.
Oestrous cyclicity (parental animals):
fresh vaginal lavage (stained with methylene blue), each morning during the pairing period, until females are mated.
Sperm parameters (parental animals):
Parameters examined in males of parental generation:
- testis weight (all groups) + microscopic evaluation (control and high-dose groups),
- epididymis weight (all groups) + microscopic evaluation (control and high-dose groups),
- microscopic evaluation of stages of the spermatogenic cycle and testicular interstitial cells (control and high-dose groups).
Litter observations:
STANDARDISATION OF LITTERS: No

PARAMETERS EXAMINED:
- number and sex of pups,
- number of live, dead and cannibalized pups,
- presence of gross anomalies, weight gain, clinical signs.

GROSS EXAMINATION OF DEAD AND SURVIVING PUPS:
- external and internal abnormalities.
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: all surviving animals after the end of the mating period after at least 5 weeks of treatment
- Female animals: all surviving animals = day 6 post-partum or, for female which had not delivered yet, day 26 post-coitum.

GROSS NECROPSY
Macroscopic post-mortem examination of principal thoracic and abdominal organs.

HISTOPATHOLOGY
- epididymides, liver, ovaries, testes, thyroid with parathyroids from all animals of the control and high-dose groups (groups 1 and 4) sacrificed at the end of the treatment period,
- liver from low- and intermediate-dose groups (groups 2 and 3) sacrificed at the end of the treatment period,
- all macroscopic lesions of all groups.

ORGAN WEIGHTS: epididymides, liver, ovaries, testes, thyroid with parathyroids.
Postmortem examinations (offspring):
SACRIFICE: on day 5 post-partum

GROSS NECROPSY: on all pups (surviving and found dead)

HISTOPATHOLOGY: No

ORGAN WEIGTHS: No
Statistics:
Body weight, food consumption and reproductive data : Data are compared by one-way analysis of variances and Dunnett test (mean values being
considered as normally distributed, variances being considered as homogenous) or by Fischer exact probability test (proportions).
Citox software (version D.6) was used to perform the statistical analysis of hematology and blood biochemistry.
PathData software (version 6.2d2) was used to perform the statistical analysis of organ weight data (level of significance: 0.05 or 0.01).
Reproductive indices:
Pre-implantation loss = 100 * (Number of corpora lutea - Number of implantation sites) / Number of corpora lutea
Post-implantation loss = 100 * (Number of implantation sites - Number of live pups) / Number of implantation sites
Mating index = 100 * (Number of mated animals / Number of paired animals)
Fertility index = 100 * (Number of pregnant female partners / Number of mated pairs)
Gestation index = 100 * (Number of females with live born pups / Number of pregnant females)
Offspring viability indices:
Live birth index = 100 * (Number of live born pups / Number of delivered pups)
Viability index on day 4 p.p. = 100 * (Number of surviving pups on day 4 p.p. / Number of live born pups)
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Ptyalism, round back and piloerection were considered to be test item-related.
Ptyalism was observed with a dose-relationship in incidence and duration in males at all dose levels. At 1000 mg/kg/day, all females were affected throughout the whole study.
Round back and piloerection were noted in males at 1000 mg/kg/day and in females at 250 and 1000 mg/kg/day. Round back was noted from day 12 or 13 of the pre-mating period and lasted 3 to 5 days at 1000 mg/kg/day. Animals treated at 1000 mg/kg/day had piloerection from day 17 (males) or 12-13 (females) of treatment and for 3-4 days (males) or up to 6 days (females). These clinical signs were also observed in females treated with 1000 mg/kg/day at the beginning of gestation only. At 250 mg/kg/day in females, piloerection and round back were noted during the premating period mainly on one day only and were thus considered not to be adverse.
To conclude, clinical signs (round back and piloerection) were considered to be adverse at 1000 mg/kg/day in male and female rats, because they were observed in the majority of the animals and lasted several days.
Four males showed loud breathing during the study, one male treated at 50 mg/kg/day and three males treated at 1000 mg/kg/day. There was no clear relationship between test item treatment and this finding: no dose-relationship as not observed at 250 mg/kg/day, and no clear pattern in appearance and duration (one out of the three animals affected at 1000 mg/kg/day experienced loud breathing for 6 days at the beginning of the treatment period, another one at the beginning of the treatment period but for 1 day only, the third male about 2 weeks later for 1 day only).
Incidental clinical signs included reflux at dosing in isolated females.
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Description (incidence):
There were no unscheduled deaths.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Males given 1000 mg/kg/day of test item had a statistically significantly lower mean body weight gain in the first week of treatment compared with controls but this was transient, had no toxicologically relevant effects on mean body weight and was therefore considered of no toxicological relevance.
There were no toxicologically relevant effects during pre-mating and lactation period in females. The same trend (at 1000 mg/kg/day) as in males in the first week of the pre-mating period was observed in females but the difference was lower than in males, with no statistical significance. During gestation, females had a lower mean body weight gain over the whole gestation period when compared with controls. As the difference was slight and did not impact significantly the mean body weight, this was considered to be of non toxicological importance.
Food consumption and compound intake (if feeding study):
effects observed, non-treatment-related
Description (incidence and severity):
Food consumption recorded in week 2 for four control males and one male of the 1000 mg/kg/day group was higher (between two to three times) than what is commonly observed for this strain of rats at about the same age (about 28 g/male/day) despite the absence of observation of food spillage. This led to high mean food consumption in the control group.
However, when excluding these abnormal data from mean calculations, mean food consumption at 1000 mg/kg/day remained strongly lower than in controls in week 2. However, this was considered to be non adverse as it had no relevant impact on mean body weight and on animal survival.
The slight higher mean food consumption noted at 250 and 1000 mg/kg/day in females during the second week of pre-mating was considered to be of doubtful toxicological importance as no statistical level, slight differences and in an opposite way compared to the effects seen in males.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
In both sexes and when compared with controls, there was a slight but statistically significant lower mean white blood cell count at 250 and 1000 mg/kg/day in males and at 1000 mg/kg/day in females, mainly due lower mean lymphocyte numbers. As the differences were slight they were considered to be non adverse and of minor toxicological importance.
In females, there were statistically significant lower mean red blood cell parameters (count, hemoglobin and/or hematocrit/PCV) from 250 mg/kg/day when compared with controls. In view of the low amplitude of differences from control values and the absence of similar findings in males, these findings were considered to be of limited toxicological importance.
The lower mean platelet levels observed in males in test item-related groups were considered to be of limited toxicological relevance as there was no dose-relationship, no similar findings in females, and the differences from the control mean were slight.
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
There was a higher mean cholesterol level from 250 mg/kg/day when compared with controls. This effect was considered to be adverse at 1000 mg/kg/day in view of the amplitude of differences from control and historical control data.
In females, there were statistically significant lower mean concentrations of inorganic phosphorus from 50 mg/kg/day and higher mean urea blood concentration at 1000 mg/kg/day. In view of the low severity of the changes and the fact that data were in the range of the historical control data, these findings were considered not to be adverse and of minor toxicological importance.
Higher mean ALAT activity was noted at 1000 mg/kg/day in females when compared with controls, while at the dose of 50 mg/kg/day a lower mean ALAT activity was observed in females and no statistical changes were observed in males in any group. Despite the fact that about half of the individual data were outside the range of the historical control data, the differences from controls and/or historical control data were slight and they were not associated with correlating adverse findings.
Higher mean calcium blood level was noted at 1000 mg/kg/day in males. All the individual data were outside the range of the historical control data, but the differences from these controls were slight.
These variations (ALAT activity and calcium blood level) were thus considered to be of limited toxicological importance and non-adverse.
The statistical significances observed for mean sodium (250 mg/kg/d in males and females), glucose (50 mg/kg/day in males), chloride (250 mg/kg/d in males) and calcium (50 and 250 mg/kg/day in females) levels were considered to be fortuitous as there was no dose-relationship, the amplitudes of variations from controls were low and/or not associated with histopathological correlates.
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
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):
Test item-related changes were confined to the liver in both sexes at 250 and 1000 mg/kg/day.
Minimal to slight centrilobular hypertrophy was observed in males given 250 mg/kg/day and in both sexes at 1000 mg/kg/day, but particularly in males. At this dose-level, this was associated with minimal or slight vacuolation of the Kupffer cells which had a foamy appearance. These changes correlated with the higher liver weights noted at necropsy and in the absence of any degenerative changes they were considered to be non-adverse.
There were no histopathological correlates with the higher mean absolute and relative thyroid weights in females or with the lower absolute epididymides weight in males.
Careful histopathological examination of testes and epididymides in males and of ovaries in females did not show any test-item related changes.

Pathology conclusion
Daily oral administration of the test item in rats at the dose-levels of 250 or 1000 mg/kg/day from before mating, through mating in males and, for females, through gestation until day 5 post partum induced centrilobular hypertrophy in males at 250 mg/kg/day and in both sexes at 1000 mg/kg/day. At this dose-level, this was associated in both sexes with vacuolation of Kupffer cells in the liver. These liver changes correlated with the higher liver weight noted at necropsy and in the absence of associated degenerative changes they were considered to be non-adverse.
Histopathological findings: neoplastic:
not examined
Other effects:
not examined
Reproductive function: oestrous cycle:
not examined
Reproductive function: sperm measures:
not examined
Reproductive performance:
no effects observed
Description (incidence and severity):
There were no test item-related effects on pairing, mating and fertility data.
All pairs mated in a maximum of 4 days except one control and one 1000 mg/kg/day pairs. In both cases, the female mated (with a second male which had already mated) after a total of 20 (control) or 16 (1000 mg/kg/day) days after the beginning of the treatment period, hence the slightly high mean pre-coital time in controls and at 1000 mg/kg/day. This was considered to be incidental as in isolated pairs per group and also in controls.
All females were pregnant except one in the control group which was sacrificed on day 26 p.c. for absence of delivery.
There were no relevant effects on duration of gestation and on number of corpora lutea and implantation sites. The gestation lasted 21 or 22 days, except for one female treated with 50 mg/kg/day (23 days).
There were no relevant effects on delivery data. The slightly higher mean number of pups delivered in controls when compared to the other groups was particularly due to one litter with 21 pups (mean of 14.4 pups delivered when excluding this litter). Three litters showed a lower number of live pups, one at 50 mg/kg/day (7 pups) and two at 1000 mg/kg/day (9 and 8 pups). It was due in two of these females to low numbers of corpora lutea (9 or 10). These findings were considered to be of no toxicological relevance as not dose-related, not severe and without significant impact on mean data.
Pre- and post-implantation losses in test item-treated groups were comparable with the individual values of the control group.
Key result
Dose descriptor:
NOAEL
Effect level:
250 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Parental toxicity.
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:
other: Reproductive performance
Remarks on result:
not determinable due to absence of adverse toxic effects
Critical effects observed:
no
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
There were no test item-related clinical signs in pups.
The most relevant clinical signs were noted in the only 3 live pups of one female (50 mg/kg/day) which were cold to the touch throughout the lactation period, and in one pup (1000 mg/kg/day) which showed several clinical signs and did not take bodyweight throughout lactation (all the others pups of the same litter didn’t show clinical signs).
This was considered to be incidental as not dose-related and in isolated litters and/or pups.
Other incidental clinical signs of pups included hematoma (hindlimb, back, head), necrosed tail, cutaneous lesions or scabs (abdomen, thorax or forelimb). The nature and incidence of these signs can be commonly observed in pups of this age or was observed with low or similar incidence as controls, and were considered not toxicologically relevant.
Dermal irritation (if dermal study):
not examined
Mortality / viability:
no mortality observed
Description (incidence and severity):
There were no test item-related deaths in pups.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
There were no effects on mean pup body weights and mean pup body weight gains.
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:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Description (incidence and severity):
There were no test item-related macroscopic findings at pup necropsy.
Histopathological findings:
not examined
Other effects:
not examined
Behaviour (functional findings):
not examined
Developmental immunotoxicity:
not examined
Key result
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
> 1 000 mg/kg bw/day
Sex:
male/female
Remarks on result:
not determinable due to absence of adverse toxic effects
Reproductive effects observed:
not specified

 Table 1 - Clinical signs:

Sex

Male

Female

Dose-level (mg/kg/day)

0

50

250

1000

0

50

250

1000

Pre-mating (females) or whole study (males)

 

 

 

 

 

. Round back

 

 

1

7

 

 

7

9

. Piloerection

 

 

 

10

 

 

2

4

. Loud breathing

 

1

 

3

 

 

 

 

. Ptyalism

1

5

10

10

 

1

 

9

Number of animals affected

1/10

6/10

10/10

10/10

0/10

1/10

7/10

10/10

Gestation

 

 

 

 

 

 

 

 

. Round back

 

 

 

 

 

 

 

1

. Piloerection

 

 

 

 

 

 

 

4

. Ptyalism

 

 

 

 

 

 

2

10

Number of animals affected

/

/

/

/

0/10

0/10

2/10

10/10

Lactation

 

 

 

 

 

 

 

 

. Ptyalism

 

 

 

 

 

 

1

10

Number of animals affected

/

/

/

/

0/9

0/10

1/10

10/10

/: not applicable.

Table 2 - Food consumption: 

Sex

Male

Female

Dose-level (mg/kg/day)

0

50

250

1000

0

50

250

1000

Pre-mating

 

 

 

 

 

 

 

 

Days 1 - 8

26

26

27

24

17

16

16

16

Days 8 -15

44

28

22*

17**

17

13

21

23

when excluding
the abnormal data
(no statistics performed)

25

28

22

10

/

/

/

/

Gestation

 

 

 

 

 

 

 

 

Days 0 - 7p.c.

/

/

/

/

21

21

21

21

Days 7 - 14p.c.

/

/

/

/

23

24

23

25

Days 14 - 20p.c.

/

/

/

/

25

26

25

25

Lactation

 

 

 

 

 

 

 

 

Days 1- 5p.p.

/

/

/

/

37

37

36

35

/: not applicable; statistically significant:*: p<0.05; **: p<0.01.

Table 3 - Hematology:  

Sex

Male

Female

Dose-level (mg/kg/day)

0

50

250

1000

0

50

250

1000

White blood cell count (G/L)

10.89

8.16

7.76*

7.79*

13.73

11.31

12.01

10.13*

Lymphocytes +
Large unstained cells (G/L)

9.34

6.96

6.28*

6.61

9.18

7.13

7.63

6.02**

Monocytes (G/L)

0.26

0.15*

0.17*

0.17*

0.56

0.44

0.41*

0.25**

Eosinophils (G/L)

0.12

0.09

0.13

0.10

0.13

0.11

0.10

0.09*

Basophils (G/L)

0.05

0.02

0.03

0.03

0.07

0.03*

0.03**

0.02**

Red blood cell count (T/L)

9.16

8.86

9.12

8.62

7.63

7.19

7.21

6.94**

Hemoglobin (g/dL)

15.8

15.2

15.9

15.3

14.2

13.6

13.4**

13.0**

Packed cell volume (PCV) (L/L)

0.49

0.47

0.49

0.47

0.44

0.42

0.41*

0.40**

Platelets (G/L)

835

656*

675*

690

1358

1302

1307

1265

Statistically significant from controls: *: p<0.05, **: p<0.01.

Table 4 - Blood biochemistry:

Sex

Male

Female

Dose-level (mg/kg/day)

0

50

250

1000

0

50

250

1000

Cholesterol (mmol/L)

1.4

1.3

2.0**

3.8**

1.6

1.7

2.1

3.6**

Inorganic phosphorus (mmol/L)

2.14

2.14

2.10

2.13

2.84

2.53*

2.49*

2.30**

Urea (mmol/L)

4.4

4.3

4.5

4.8

6.5

6.4

7.2

8.1*

Alanine aminotransferase (ALAT) (IU/L)

41

45

42

45

86

65**

78

117**

Calcium (mmol/L)

2.74

2.70

2.77

2.93**

2.95

2.83*

2.80**

2.95

Glucose (mmol/L)

7.72

8.79*

7.89

7.69

6.35

6.47

6.54

6.02

Sodium (mmol/L)

149.0

149.6

150.9**

149.8

142.8

144.6

145.0*

143.7

Chloride (mmol/L)

103.9

104.8

105.7*

105.3

98.9

100.3

100.1

99.5

Statistically significant from controls: *: p<0.05; **: p<0.01.

Table 5 - Organ weights:  

Sex

Male

Female

Group

2

3

4

2

3

4

Dose-level (mg/kg/day)

50

250

1000

50

250

1000

Number of animals

10/10

10/10

10/10

10/10

10/10

10/10

Body weight

0

-2

-8

+3

+1

-4

- Liver

 

 

 

 

 

 

  . absolute

+6

+11

+19*

+3

+5

+16*

  . relative

+6

+14**

+29**

+1

+4

+22**

- Thyroids

 

 

 

 

 

 

  . absolute

-6

+2

-14

+5

+24

+31

  . relative

-6

+4

-6

+2

+22

+37*

- Epididymides

 

 

 

 

 

 

  . absolute

-3

-3

-10*

 

 

 

  . relative

-2

-1

-2

 

 

 

Statistically significant from controls: *: p<0.05, **: p<0.01

The significance concerned the organ weights values and not the percentages.

Table 6 - Microscopic examination: 

Sex

Males

Females

Group

1

2

3

4

1

2

3

4

Dose-level (mg/kg/day)

0

50

250

1000

0

50

250

1000

Number of animals

10

10

10

10

10

10

10

10

Hypertrophy; hepatocytes

Minimal

-

-

6

6

-

-

-

1

Slight

-

-

-

4

-

-

-

2

Total

0

0

6

10

0

0

0

3

Vacuolation, Kuffer cells

Minimal

-

-

-

8

-

-

-

3

Slight

-

-

-

2

-

-

-

-

Total

0

0

0

10

0

0

0

3

-: not observed.

Conclusions:
The test item was administered daily by oral gavage to male and female Sprague Dawley rats, for 2 weeks before pairing, during pairing, gestation and until day 5 p.p., at dose-levels of 50, 250 or 1000 mg/kg/day.
At 1000 mg/kg/day, round back and piloerection noted in both sexes in weeks 2/3, as well as the higher mean cholesterol level at the end of the treatment period were considered to be adverse.
There were no adverse findings at 50 and 250 mg/kg/day.

Therefore, based on the experimental conditions of this study:
- the No Observed Adverse Effect Level (NOAEL) for parental toxicity was considered to be 250 mg/kg/day,
- the NOAEL for reproductive performance (mating and fertility) was considered to be 1000 mg/kg/day,
- the NOAEL for toxic effects on progeny was considered to be 1000 mg/kg/day.
Executive summary:

The objective of this study was to evaluate the potential toxic effects of the test item following daily oral administration (gavage) to male and female rats from before mating, through mating and, for females, through gestation until day 5 post-partum (p.p.), based on the OECD guideline No. 421, 27 July 1995.

This study provides information on male and female reproductive performance, such as gonadal function, mating behavior, conception, development of the conceptus and parturition.

The study was conducted in compliance with the principles of Good Laboratory Practice Regulations.

 

Methods

Three groups of ten male and ten female Sprague-Dawley rats received the test item daily by oral (gavage) from before pairing, through pairing and, for the females, through gestation until day 5 p.p. The test item was administered as a suspension in the vehicle,corn oil, at dose-levels of 50, 250 or 1000 mg/kg/day.

Another group of ten males and ten females received the vehicle alone under the same experimental conditions and acted as a control group. A constant dosage-volume of 5 mL/kg/day was used.

 

The concentration of the dose formulation was checked in study weeks 1, 3 and 6.

 

The animals were checked at least twice daily during the dosing period for mortality and morbidity and once daily for clinical signs. Body weight and food consumption were recorded at leastonce a week. The animals were paired for mating after 2 weeks of treatment and the dams were allowed to litter and rear their progeny until day 5 p.p. The litter sizes and numbers of pups per sex were recorded after birth. The pups were observed daily for clinical signs, abnormal behaviour and external abnormalities, and weighed on days 1 and 5 p.p.

Hematology and blood biochemistry were carried outfor all males and all females sacrificed on day 6 p.p.

The males were sacrificed after 5 weeks of treatment and the dams on day 6 p.p. Final body weights and selected organs weights (epididymides, liver, ovaries, testes, thyroid with parathyroids) were recorded and a macroscopic post-mortem examinationof the principal thoracic and abdominal organswas performed, with particular attention paid to the reproductive organs. A microscopic examination was performed on selected organs from the control and high-dose groups (epididymides, liver, testes, thyroid with parathyroids), on the liver from low- and intermediate-dose groups and on all macroscopic lesions.

The pups were sacrificed on day 5 p.p. and submitted for a macroscopic post-mortem examination of the principal thoracic and abdominal organs.

 

Results

 

The test item concentrations in the administered dose formulations analyzed in weeks 1, 3 and 6 were within the acceptance criteria (± 15% of the nominal concentrations) and no test item was found in control samples.


There were no unscheduled deaths. Ptyalism was observed in males at all dose-levels (half of the animals at 50 mg/kg/day, all animals at 250 and 1000 mg/kg/day) and in all females at 1000 mg/kg/day. Round back was noted from the end of week 2 in females (7/10) at 250 mg/kg/day and in both sexes at 1000 mg/kg/day (7/10 males, 9/10 females). Piloerection was also observed at 250 mg/kg/day in females (2/10) and at 1000 mg/kg/day in all males and 4/10 females. At 1000 mg/kg/day, round back and piloerection lasted 3 to 6 days and were considered to be adverse. During gestation, one female still had round back on the first day of gestation, and four females piloerection at the beginning of gestation only.

There were no toxicologically significant effects on mean body weight gain and mean body weight. When compared with controls, mean food consumption at 1000 mg/kg/day was lower in males; this change was considered not to be adverse because of no relevant impact on mean body weight and on animal survival.

 

At hematology, there was a slight lower mean white blood cell count than in controls at 250 and 1000 mg/kg/day in males and at 1000 mg/kg/day in females, mainly related to lower mean numbers of lymphocytes. This was considered to be non adverse and of minor toxicological importance.

At blood biochemistry and when compared with controls, there was a dose-related higher mean cholesterol level from 250 mg/kg/day. This effect was considered to be adverse at 1000 mg/kg/day in view of the amplitude of differences from control and historical control data. In females, there was also a dose-related lower mean concentration of inorganic phosphorus from 50 mg/kg/day and a slightly higher mean urea blood concentration at 1000 mg/kg/day. These changes in mean inorganic phosphorus and urea levels were considered not to be adverse and of minor toxicological importance.

 

There were no test item-related macroscopic changes or effects on mean thyroid/parathyroid, epididymides, testis and ovary weights at necropsy. The microscopic examination and mean organ weights showed test item-related changes in liver only. Administration of the test item in rats at 1000 mg/kg/dayinduced mild centrilobular hypertrophy and vacuolation of Kupffer cells in the liver which correlated with the higher mean liver weight at necropsy. Centrilobular hypertrophy alone was also observed in males at 250 mg/kg/day. In the absence of any associated degenerative liver changes, these observations were considered not to be adverse.

 

There were no test item-related effects on pairing, mating and fertility data. There were no toxicologically relevant effects on delivery data.

 

In pups, there were no test item-related deaths or clinical signs andno test item-related effects on mean pup body weights, mean pup body weight gains,on the percentage of male pups at birth and no test item-related findings noted at necropsy.

 

Conclusion

The test item was administered daily by oral gavage to male and female Sprague-Dawley rats, for 2 weeks before pairing, during pairing, gestation and until day 5p.p., at dose-levels of 50, 250 or 1000 mg/kg/day.

 

At 1000 mg/kg/day,round back and piloerection noted in both sexes in weeks 2/3, as well as the higher mean cholesterol level at the end of the treatment period were considered to be adverse.

There were no adverse findings at 50 and 250 mg/kg/day.

 

Therefore, based on the experimental conditions of this study:

.         the No Observed Adverse Effect Level (NOAEL) for parental toxicity was considered to be 250 mg/kg/day,

.         the NOAEL for reproductive performance (mating and fertility) was considered to be 1000 mg/kg/day,

.         the NOAEL for toxic effects on progeny was considered to be 1000 mg/kg/day.

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
This screening test is considered to be reliable: study was performed according to guideline study and has a klimisch score of 1.
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

Screening test on reproduction (Bentz 2013):

The objective of this study was to evaluate the potential toxic effects of the test item following daily oral administration (gavage) to male and female rats from before mating, through mating and, for females, through gestation until day 5 post-partum (p.p.), based on the OECD guideline No. 421, 27 July 1995. This study provides information on male and female reproductive performance, such as gonadal function, mating behavior, conception, development of the conceptus and parturition.

Three groups of ten male and ten female Sprague-Dawley rats received the test item daily by oral (gavage) from before pairing, through pairing and, for the females, through gestation until day 5p.p. The test item was administered as a suspension in the vehicle, corn oil, at dose-levels of 50, 250 or 1000 mg/kg/day. Another group of ten males and ten females received the vehicle alone under the same experimental conditions and acted as a control group. A constant dosage-volume of 5 mL/kg/day was used.

The animals were checked at least twice daily during the dosing period for mortality and morbidity and once daily for clinical signs. Body weight and food consumption were recorded at leastonce a week. The animals were paired for mating after 2 weeks of treatment and the dams were allowed to litter and rear their progeny until day 5 p.p.The litter sizes and numbers of pups per sex were recorded after birth. The pups were observed daily for clinical signs, abnormal behaviour and external abnormalities, and weighed on days 1 and 5p.p.

Hematology and blood biochemistry were carried out for all males and all females sacrificed on day 6p.p.

The males were sacrificed after 5 weeks of treatment and the dams on day 6p.p.Final body weights and selected organs weights (epididymides, liver, ovaries, testes, thyroid with parathyroids) were recorded and a macroscopicpost-mortemexaminationof the principal thoracic and abdominal organswas performed, with particular attention paid to the reproductive organs. A microscopic examination was performed on selected organs from the control and high-dose groups (epididymides, liver, testes, thyroid with parathyroids), on the liver from low- and intermediate-dose groups and on all macroscopic lesions. The pups were sacrificed on day 5 p.p. and submitted for a macroscopicpost-mortem examination of the principal thoracic and abdominal organs

There were no unscheduled deaths. Ptyalism was observed in males at all dose-levels (half of the animals at 50 mg/kg/day, all animals at 250 and 1000 mg/kg/day) and in all females at 1000 mg/kg/day. Round back was noted from the end of week 2 in females (7/10) at 250 mg/kg/day and in both sexes at 1000 mg/kg/day (7/10 males, 9/10 females). Piloerection was also observed at 250 mg/kg/day in females (2/10) and at 1000 mg/kg/day in all males and 4/10 females. At 1000 mg/kg/day, round back and piloerection lasted 3 to 6 days and were considered to be adverse. During gestation, one female still had round back on the first day of gestation, and four females piloerection at the beginning of gestation only.

There were no toxicologically significant effects on mean body weight gain and mean body weight. When compared with controls, mean food consumption at 1000 mg/kg/day was lower in males; this change was considered not to be adverse because of no relevant impact on mean body weight and on animal survival.

At hematology, there was a slight lower mean white blood cell count than in controls at 250 and 1000 mg/kg/day in males and at 1000 mg/kg/day in females, mainly related to lower mean numbers of lymphocytes. This was considered to be non adverse and of minor toxicological importance.

At blood biochemistry and when compared with controls, there was a dose-related higher mean cholesterol level from 250 mg/kg/day. This effect was considered to be adverse at 1000 mg/kg/day in view of the amplitude of differences from control and historical control data. In females, there was also a dose-related lower mean concentration of inorganic phosphorus from 50 mg/kg/day and a slightly higher mean urea blood concentration at 1000 mg/kg/day. These changes in mean inorganic phosphorus and urea levels were considered not to be adverse and of minor toxicological importance.

There were no test item-related macroscopic changes or effects on mean thyroid/parathyroid, epididymides, testis and ovary weights at necropsy. The microscopic examination and mean organ weights showed test item-related changes in liver only. Administration of the test item in rats at 1000 mg/kg/dayinduced mild centrilobular hypertrophy and vacuolation of Kupffer cells in the liver which correlated with the higher mean liver weight at necropsy. Centrilobular hypertrophy alone was also observed in males at 250 mg/kg/day. In the absence of any associated degenerative liver changes, these observations were considered not to be adverse.

There were no test item-related effects on pairing, mating and fertility data. There were no toxicologically relevant effects on delivery data.

In pups, there were no test item-related deaths or clinical signs andno test item-related effects on mean pup body weights, mean pup body weight gains,on the percentage of male pups at birth and no test item-related findings noted at necropsy.

Finally, at 1000 mg/kg/day,round back and piloerection noted in both sexes in weeks 2/3, as well as the higher mean cholesterol level at the end of the treatment period were considered to be adverse. There were no adverse findings at 50 and 250 mg/kg/day. 

Therefore, based on the experimental conditions of this study: the No Observed Adverse Effect Level (NOAEL) for parental toxicity was considered to be 250 mg/kg/day, the NOAEL for reproductive performance (mating and fertility) and  the NOAEL for toxic effects on progeny were considered to be 1000 mg/kg/day

Effects on developmental toxicity

Description of key information
  
    
    

A developmental study on rat by oral route has been performed: Ethoxylated bisphenol A diacrylate was administered by gavage, once daily, from Days 6 to 20p.c.inclusive, to pregnant Sprague-Dawley rats at dosages of 50, 250 and 1000 mg/kg/day.

At 1000 mg/kg/day, lower fetal body weight (female fetuses) and increased fetal ossification delays were observed in presence of some signs of limited maternal toxicity (i.e. lower body weight gain and net body weight gain from Day 6p.c.). At 50 and 250 mg/kg/day, there were signs of slight fetal ossification delays.

Under the experimental conditions and results of this study: the No Observed Adverse Effect Level (NOAEL) for maternal parameters and for embryo-fetal development were considered to be 1000 mg/kg/day.

 

 

 

 

Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
03 March 2017 - 20 April 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Version / remarks:
22 January 2001
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: breeder: Janvier, le Genest-Saint-Isle, France
- Age at the beginning of the treatment period: 10-11 weeks old
- Mean body weight at the beginning of the treatment period: a mean body weight of 315 g (range: 232 g to 360 g)
- Fasting period before study: no
- Housing: The animals were individually housed in polycarbonate cages (Tecniplast 2154, 940 cm2)
- Diet: SSNIFF R/M-H pelleted diet (free access)
- Water: tap water filtered with a 0.22 µm filter (free access)
- Acclimation period: for a period of 4 or 5 days before the beginning of the treatment period.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C
- Humidity (%): 50 ± 20%
- Air changes (per hr): about 8 to 15 cycles/hour of filtered, non-recycled air
- Photoperiod (hrs dark / hrs light): 12 h/12 h

IN-LIFE DATES: 27 March 2017 to 20 April 2017.
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
PREPARATION OF DOSING FORMULATIONS:
- Emulsion in the vehicle
- Concentration in vehicle: 10, 50 and 200 mg/mL
- Amount of vehicle (if gavage): 5 mL/kg/day
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Type of method: High Performance Liquid Chromatography with UV detection (HPLC/UV)
Test item concentrations:remained within an acceptable range of variation (-4.2% to +2.6%) when compared to the nominal values.
Homogeneity: the dose formulations containing the test item in corn oil at 2 and 200 mg/mL were found to be homogeneous. They are therefore considered to be suitable for routine administration in GLP Toxicological studies, based on a daily preparation.
Stability: no stability available, dose formulations prepared daily.
Details on mating procedure:
The females were mated at the breeder's facilities. The day of confirmed mating (detection of a vaginal plug) was designated as Day 0 p.c.
Duration of treatment / exposure:
The dose formulations were administered daily from Days 6 to 20 p.c., inclusive.
Frequency of treatment:
Daily
Dose / conc.:
0 mg/kg bw/day (actual dose received)
Dose / conc.:
50 mg/kg bw/day (actual dose received)
Dose / conc.:
250 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:
24 females (groups 2, 3 and 4).
For group 1 (control group), at animal receipt, one female presented a malformation at the inferior lip. This female was excluded from the study.
Only 23 females instead of 24 were therefore assigned to the control group.
Control animals:
yes, concurrent vehicle
Details on study design:
- Rationale for dose level selection:
The dose levels were selected in agreement with the Sponsor, on the basis of the results of a previous OECD 421 study (5 weeks of treatment for males, about 6 weeks for females in total) performed in the same species and strain at 50, 250 and 1000 mg/kg/day. In females of this study, there were no unscheduled deaths and no toxicologically significant effects on mean body weight, mean food consumption or mean reproductive and litter parameters. However, adverse findings were noted in females at 1000 mg/kg/day: transient round back and piloerection in Weeks 2/3 and higher mean cholesterol level at the end of the treatment period.

- Rationale for animal assignment: computerized stratification procedure.
Maternal examinations:
MORTALITY/MORBIDITY:
- Time schedule: each animal was checked for mortality and morbidity once a day before the treatment period and at least twice a day during the treatment period, including weekends and public holidays.

CLINICAL SIGNS:
- Time schedule: from arrival, each animal was observed once a day as part of the routine examinations. From the start of the treatment period, each animal was observed once a day, at approximately the same time, for the recording of clinical signs.

BODY WEIGHT:
- Time schedule: the body weight of each female was recorded on Days 2, 4, 6, 9, 12, 15, 18 and 21 p.c.

FOOD CONSUMPTION:
- Time schedule: the quantity of food consumed by each female was recorded for the following intervals: Days 2-4, 4-6, 6-9, 9-12, 12-15, 15-18 and 18-21 p.c.

POST-MORTEM MACROSCOPIC EXAMINATION:
- Sacrifice on Day 21 post-coitum.
- Examined: principal thoracic and abdominal organs.
Ovaries and uterine content:
The ovaries and uterine content were examined after termination, including:
- gravid uterus weight,
- number of corpora lutea,
- number and distribution of dead and live fetuses,
- number and distribution of early and late resorptions,
- number and distribution of uterine scars,
- number and distribution of implantation sites,
- gross evaluation of placentas,
- placental weight.
Fetal examinations:
- External examinations: Yes: all fetuses per litter
- Soft tissue examinations: Yes: half fetuses per litter
- Skeletal examinations: Yes: half fetuses per litter
- Head examinations: Yes: half fetuses per litter
- Other: body weight, sex
Statistics:
Data were compared by one-way analysis of variance and Dunnett test (mean values being considered as normally distributed and variances being considered as homogeneous) or by Fisher exact probability test (proportions).
Indices:
% Pre-implantation loss = 100 * (Number of corpora lutea - Number of implantation sites) / Number of corpora lutea
% Post-implantation loss = 100 * (Number of implantation sites - Number of live fetuses) / Number of implantation sites
Historical control data:
Cf attached document
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
See table 1.
At 50 mg/kg/day, one female had a mass on mammary gland from Day 16 p.c. As this observation was limited to one animal from the low-dose group only, a relationship to the test item treatment was considered unlikely.

At 250 mg/kg/day, there were no clinical signs.

At 1000 mg/kg/day, almost all the females had ptyalism for 1 to 13 days of treatment. This test item-related finding was considered as non-adverse (not critical, sign of discomfort).
One female started showing round back and piloerection on Day 19 p.c. and reddish vaginal discharge was found on Day 21 p.c. This animal also had a strongly reduced food consumption from Day 15 p.c. and lost 15% of body weight from Days 15 to 18 p.c. with a low body weight gain thereafter. At hysterectomy, only implantation scars were found and at necropsy the female had a segmental aplasia of the right uterine horn and no right kidney/ureter (congenital anomalies), as well as an enlarged spleen. In absence of similar findings on the clinical condition of the other females of the group and in presence of the congenital anomalies in this female, a test item effect was considered to be unlikely.
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Description (incidence):
No unscheduled death occurred during the study.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
See table 2.
At 50 and 250 mg/kg/day, there were no effects on mean body weight and mean body weight change.
At 1000 mg/kg/day, there was a lower mean body weight gain vs. controls at the beginning of the treatment (Days 9 to 12 p.c., p<0.001). This effect were considered to be test item treatment-related and non adverse, in absence of effect on mean body weight and in view of the slight and transient effect on mean body weight change. It correlated with the effects on mean net body weight change.
Food consumption and compound intake (if feeding study):
effects observed, non-treatment-related
Description (incidence and severity):
See table 3.
There were no effects on mean food consumption, except a slightly higher mean food consumption at 1000 mg/kg/day between Days 18 and 21 p.c. (p<0.05). In view of the limited difference from controls, this was considered of no biologically significance and unlikely test item-related.
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
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
See table 4.
None of the necropsy findings were ascribed to the test item treatment: they were noted in isolated cases, were not dose-related and/or were congenital anomalies.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Other effects:
effects observed, treatment-related
Description (incidence and severity):
Mean gravid uterus and carcass weights, and mean net body weight changes
See table 5.
At 50 and 250 mg/kg/day, there were no effects on mean carcass and gravid uterus weights and on mean net body weight change from Day 6 p.c.
At 1000 mg/kg/day, the mean net body weight change from Day 6 p.c. was lower than in controls (p<0.05), correlating with the effects on mean body weight change. This effect was considered to be test item-related and non-adverse (no toxicologically significant effects on mean carcass weight and slight difference) and test item treatment-related. There were no effects on mean gravid uterus weight.
Number of abortions:
not examined
Pre- and post-implantation loss:
effects observed, non-treatment-related
Description (incidence and severity):
See table 6.
There were no test item treatment-related findings.
Total litter losses by resorption:
effects observed, non-treatment-related
Description (incidence and severity):
See table 6.
There were no test item treatment-related findings.

At 250 mg/kg/day, the higher mean number of late resorptions when compared with controls was due to one female with 100% of implantation loss.
At 1000 mg/kg/day, the higher mean number of implantation scars when compared with controls was due to one female only (with implantation scars only).
Early or late resorptions:
effects observed, non-treatment-related
Description (incidence and severity):
See table 6.
There were no test item treatment-related findings.

At 250 mg/kg/day, the higher mean number of late resorptions when compared with controls was due to one female with 100% of implantation loss.
At 1000 mg/kg/day, the higher mean number of implantation scars when compared with controls was due to one female only (with implantation scars only).
Dead fetuses:
no effects observed
Changes in pregnancy duration:
not examined
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): not examined
Changes in number of pregnant:
effects observed, non-treatment-related
Description (incidence and severity):
See table 6.
On Day 21 p.c., there were 20/23, 23/24, 20/24 and 22/24 females pregnant with live fetuses at 0, 50, 250 and 1000 mg/kg/day, respectively.
The other females were non-pregnant with the exception of one female at 250 mg/kg/day and one at 1000 mg/kg/day which had implantation scars/total resorptions.
Other effects:
not specified
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
body weight and weight gain
Fetal body weight changes:
effects observed, treatment-related
Description (incidence and severity):
See table 7.
At 1000 mg/kg/day when compared with controls, female fetuses had a lower mean body weight (p<0.01), which was included in Historical Control Data. This slight effect was considered to be test item-related and non-adverse.
Similar trend was observed in males at the same dose or in females at the mid-dose but without statistical significance and within the range of the Historical Control range. This trend was therefore considered to be not toxicologically significant.

Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): not examined
Reduction in number of live offspring:
not examined
Changes in sex ratio:
no effects observed
Description (incidence and severity):
See table 8.
There were no effects on mean sex ratio.
Changes in litter size and weights:
no effects observed
Description (incidence and severity):
See table 6.
There were no test item treatment-related findings.
Changes in postnatal survival:
not examined
External malformations:
effects observed, non-treatment-related
Description (incidence and severity):
Variations
See table 10.
There were no external variations in fetuses at 50 and 250 mg/kg/day.
At 1000 mg/kg/day one litter had three fetuses with small mandible and protruding tongue. They were associated with cleft palate. Considering that these variations were observed in only one litter and were also observed in Historical Control Data at similar litter incidence, a relationship to the test-item was considered unlikely.

Malformations
See table 11.
At 50 mg/kg/day, one fetus displayed various severe external malformations. Taking into account that these malformations affected only one fetus, were observed in controls or Historical Control Data for most of them and did not occur in mid-dose or high-dose group, this finding was considered to be not test item treatment-related.
At 250 mg/kg/day, there were no fetal external malformations.
At 1000 mg/kg/day, three fetuses from the same litter had a cleft palate (and small mandible/protruding tongue, see above). Considering that this malformation was observed in only one litter and was also observed in Historical Control Data at similar litter incidence, it was considered to be not test item treatment-related.
Skeletal malformations:
effects observed, treatment-related
Description (incidence and severity):
Cartilages
There were no test item treatment-related findings at cartilage examinations.
All cartilages were present and had a normal morphology with the exceptions of one fetus at 250 mg/kg/day (absent rib) and another one at 1000 mg/kg/day (misshapen carpal joint). These findings were considered to be not test item treatment-related.

Variations
See table 14.
At 1000 mg/kg/day and when compared with controls, there were higher fetal and litter incidences of incomplete ossification of cervical and thoracic vertebrae centra (including bipartite and dumbbell ossification of thoracic vertebrae centra) and of unossification of the 1st metatarsal bones. Ossification of metacarpal bones and caudal vertebrae centra also had the tendency to be delayed. The fetal and litter incidences of unossified 1st metatarsal bone and of unossified/incomplete ossified metacarpal(s) were outside the Historical Control Data (as well as the litter incidence of bipartite ossification of thoracic vertebrae and the fetal incidence of incompletely ossified caudal vertebrae centra).

Higher litter and fetal incidences of some of these bones tended also, and in a dose-related manner, to be higher at 50 and 250 mg/kg/day than in controls but they remained within the Historical Control Data.

These ossification delays were considered to be test item-related and non-adverse, associated with a slight effect on mean fetal body weight at 1000 mg/kg/day..

Other differences from controls were considered as not test item treatment-related as they were isolated, recorded at lower/similar incidences than/as in control groups and/or HCD, and/or were not dose-related.
The incomplete ossifications noted on head, hindlimb and forlimb bones described in the table above were only observed in 2 fetuses from the same litter already mentioned.

Malformations
See table 15.
At 50 and 250 mg/kg/day, there were no skeletal malformations.

At 1000 mg/kg/day, one fetus had head (absent nasal bone and split palate) and hindpaw malformations (unossified tibia and fibula), as well as another fetus from the same litter (split palate, short tibia and short fibula). One fetus from another litter displayed supernumerary digit(s) (right forepaw).
Almost all skeletal malformations noted at 1000 mg/kg/day were observed in 2 fetuses from the same litter. The litter incidence of split palate noted in both these fetuses was within Historical Control Data, and a third fetus from the same litter was also affected (see external malformations, skeleton not examined) while there was no other litter in the group with split/cleft palate. In addition, these 2 fetuses also had ossification delays on head, limb and/or metatarsal bones (variations), and/or misshapen carpal joint, which all were also not seen in any other fetuses from the study.
Therefore, malformations noted in these fetuses from the same litter were likely due to a litter bias.
The digit malformation of the fetus from another litter was not similar (different morphological pattern) to the limb malformations in litter mentioned above, and despite the fact that it can not be found in our Historical Control Data, it can occur spontaneously in laboratory rats (Lang 1993, Morita et al. 1987).
Therefore, a relationship between skeletal malformations in litters at 1000 mg/kg/day and the test item treatment was considered to be unlikely.
Visceral malformations:
effects observed, non-treatment-related
Description (incidence and severity):
Variations
See table 12.
None of the soft tissue variations were considered to be related to the test item treatment: they were not dose-related and/or recorded at incidences lower than Historical Control Data and/or controls.

Malformations
See table 13.
None of the soft tissue malformations were considered to be related to the test item treatment: they were not dose-related, were noted in one same litter only and/or had similar incidences as Historical Control Data.
Other effects:
no effects observed
Description (incidence and severity):
Placental weight
See table 9.
There were no effects on mean placental weight.
Details on embryotoxic / teratogenic effects:
There were no toxicologically significant differences in the incidence of litters with malformed fetuses between groups, nor any dose-relationship in the incidence of each malformation.
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
fetal/pup body weight changes
other: skeletal variations
Abnormalities:
effects observed, non-treatment-related
Developmental effects observed:
no

Table 1: Clinical signs

 

Dose level (mg/kg/day)

0

50

250

1000

Ptyalism

 

 

 

20

Piloerection

 

 

 

1a

Round back

 

 

 

1a

Reddish vaginal discharge

1

 

 

1a

Mass on mammary gland

 

1

 

 

Scab on abdomen or neck

1

 

 

1

No remarkable observations

18/20

22/23

21/21

3/23

a: same female.

Table 2: Body weight

 

Dose level (mg/kg/day)

0

50

250

1000

Body weight

 

 

 

 

Day 6p.c.

317

317

316

314

 

 

(0)

(0)

(-1)

Day 9p.c.

328

330

327

322

 

 

(+1)

(0)

(-2)

Day 12p.c.

351

353

348

337

 

 

(+1)

(-1)

(-4)

Day 21p.c.

462

469

469

445

 

 

(+2)

(+2)

(-4)

Body weight change

 

 

 

 

Days 6 - 9p.c.

+11

+13

+11

+8

Days 9 - 12p.c.

+22

+22

+21

+15#

Days 6 - 21p.c.

+145

+152

+154

+131*

Statistical significance: *:p<0.05;#: p<0.001.

(): % from controls.

Table 3: Food consumption

 

Dose level (mg/kg/day)

0

50

250

1000

Days 6 - 9p.c.

22

23

23

22

 

 

(+5)

(+5)

(0)

Days 9 - 12p.c.

24

25

25

23

 

 

(+4)

(+4)

(-4)

Days 12 - 15p.c.

26

27

28

27

 

 

(+4)

(+8)

(+4)

Days 15 - 18p.c.

28

29

30

29

 

 

(+4)

(+7)

(+4)

Days 18 - 21p.c.

30

30

31

32*

 

 

(0)

(+3)

(+7)

Statistical significance:*:p<0.05.

(): % from controls.

Table 4: Maternal necropsy findings

 

Dose level (mg/kg/day)

0

50

250

1000

Spleen: enlarged

 

 

1a

1b

Liver: reduced in size + mass on median lobe

 

1

 

 

Mammary gland: palpable mass

 

1

 

 

Vagina: colored contents

 

 

1a

 

Placenta(s): enlarged

1

 

1

1

Colored material around placenta

 

 

 

1

Placentas fused

1

 

 

1

Kidney: enlarged (unilateral)

 

 

1c

 

Kidney: absence (unilateral)

 

1d

1c

1b

Ureter: absence (unilateral)

 

1d

1c

1b

Uterus: absence of uterine horn (unilateral)

 

1d

1c

 

Uterus: segmental aplasia of uterine horn (unilateral)

 

 

 

1b

No remarkable observations

18/20

20/23

18/21

19/23

a: same female, total resorption.

b: same female, implantation scars only.

c: same female.

d: same female.

Table 5: Gravid uterus weight, carcass weight and net body weight change

 

Dose level (mg/kg/day)

0

50

250

1000

Gravid uterus weight

93.3

96.9

100.2

96.8

 

 

(+4)

(+7)

(+4)

Carcass weight

368.4

372.1

369.0

348.1

 

 

(+1)

(0)

(-6)

Net body weight change from Day 6p.c.

+51.5

+54.9

+53.4

+33.9**

Statistical significance: **: p<0.01.

(): % from controls.

Table 6: Hysterectomy data

 

Dose level (mg/kg/day)

0

50

250

1000

Number of females with live fetuses

20

23

20

22

Mean number ofcorpora lutea

14.9

15.7

15.8

15.2

Mean number of implantation sites

12.4

12.8

13.0

12.8

Mean pre-implantation loss (%)

17.5

18.0

16.8

15.5

Mean number of fetuses

11.1

11.9

11.8

11.5

Mean dead fetuses (%)

0.0

0.0

0.0

0.0

Mean number of implantation scars

0.0

0.0

0.0

0.4

Mean number of early resorptions

1.0

0.8

0.6

0.8

Mean number of late resorptions

0.2

0.1

0.7

0.1

Mean post-implantation loss (%)

10.1

7.7

8.3

11.1

 

Table 7: Fetal body weight

 

Dose level (mg/kg/day)

0

50

250

1000

HCD

Mean fetal body weight
(males and females)

5.91

5.80

5.77

5.64

[5.5-5.9]

 

 

(-2)

(-2)

(-5)

 

males

6.06

5.97

5.99

5.83

[5.7-6.1]

 

 

(-1)

(-1)

(-4)

 

females

5.77

5.64

5.55

5.44**

[5.4-5.7]

 

 

(-2)

(-4)

(-6)

 

(): % from controls.

**: p<0.01

HCD: Historical Control Data (Sprague-Dawley rats from the Supplier Janvier; signed in December 2016): [min study mean; max study mean].

Table 8: Fetal sex ratio

 

Dose level (mg/kg/day)

0

50

250

1000

Mean percentage of male fetuses

47.9

50.3

48.7

51.4

Table 9: Placental weight

 

Dose level (mg/kg/day)

0

50

250

1000

Mean placental weight

0.69

0.68

0.69

0.69

 

Table 10: Fetal external variations

 

Dose level (mg/kg/day)

0

50

250

1000

HCD

Number of litters

20

23

20

22

388

Number of fetuses

223

273

247

265

5000

Small mandible, F (L)

1.1 (4.5)a

0.4 (5.0)b

Protruding tongue, F (L)

1.1 (4.5)a

0.4 (4.8)

Litters with external variation, n (%)

1 (4.5)

6 (1.5)

Fetuses with external variation, n (%)

 

3 (1.1)

7 (0.1)

F: fetal incidence (%).

L: litter incidence (%).

a: same fetuses;b: referred as short mandible in HCD.

HCD: Historical Control Data (Sprague-Dawley rats from the Supplier Janvier; signed in December 2016): max fetal incidence (max litter incidence) for the findings.

 

Table 11: Fetal skeletal malformations

 

Dose level (mg/kg/day)

0

50

250

1000

HCD

Number of litters

20

23

20

22

388

Number of fetuses

117

143

131

138

2596

Nasal: absent

 

 

 

0.7 (4.5)a

-

Palate: split

 

 

 

1.4 (4.5)a, b

0.7 (5.0)

Fused sternebra(e)

0.9 (5.0)

 

 

 

1.3 (8.7)

Absent sternebra(e)

0.9 (5.0)

 

 

 

-

Forepaw: supernumerary digit(s)

 

 

 

0.7 (4.5)

-

Tibia: unossified (unilateral)

 

 

 

0.7 (4.5)a

-

Tibia: short

 

 

 

0.7 (4.5)b

-

Fibula: unossified (unilateral)

 

 

 

0.7 (4.5)a

-

Fibula: short

 

 

 

0.7 (4.5)b

-

Litters with skeletal malformation, n (%)

1 (5.0)

0 (0.0)

0 (0.0)

2 (9.1)

15 (3.9)

Fetuses with skeletal malformation, n (%)

1 (0.9)

0 (0.0)

0 (0.0)

3 (2.2)

18 (0.7)

F: fetal incidence; L: litter incidence.

a and b: two fetuses from the same litter.

HCD: Historical Control Data (Sprague-Dawley rats from the Supplier Janvier; signed in December 2016): max fetal incidence (max litter incidence) for the findings; -: none in HCD.

Table 12: Fetal soft tissue variations

Dose level (mg/kg/day)

0

50

250

1000

HCD

Number of litters

20

23

20

22

387

Number of fetuses

106

130

116

127

2404

Dilated cerebral ventricle, F (L)

0.9 (5.0)

 

 

 

0.8 (4.8)

Enlarged atrial chamber, F (L)

 

0.8 (4.3)

 

 

-

Malpositioned subclavian artery, F (L)

 

 

0.9 (5.0)

 

-

Short innominate artery, F (L)

3.8 (15.0)

1.5 (8.7)

4.3 (10.0)

0.8 (4.5)

3.7 (22.7)

Absent innominate artery, F (L)

 

 

 

2.4 (9.1)

5.1 (25.0)

Lung: colored area, F (L)

 

0.8 (4.3)

 

 

-

Dilated renal pelvis, F (L)

 

1.5 (8.7)

 

2.4 (13.6)

9.5 (28.6)

Dilated uterine horn, F (L)

 

 

 

0.8 (4.5)

-

Dilated ureter, F (L)

0.9 (5.0)

1.5 (8.7)

 

1.6 (9.1)

7.1 (28.0)

Litters with soft tissue variation, n (%)

4 (20.0)

6 (26.1)

3 (15.0)

7 (31.8)

86 (22.2)

Fetuses with soft tissue variation, n (%)

6 (5.7)

6 (4.6)

6 (5.2)

9 (7.1)

119 (5.0)

F: fetal incidence; L: litter incidence.

HCD: Historical Control Data (Sprague-Dawley rats from the Supplier Janvier; signed in December 2016): max fetal incidence (max litter incidence) for the findings; -: none in HCD.

Table 13: Fetal soft tissue malformations

 

Dose level (mg/kg/day)

0

50

250

1000

HCD

Number of litters

20

23

20

22

387

Number of fetuses

106

130

116

127

2404

Cleft palate, F (L)

 

 

 

0.8 (4.5)

b

Absent kidney, F (L)

 

 

1.7 (5.0)a

 

0.7 (4.5)

Absent ureter, F (L)

 

 

1.7 (5.0)a

 

-

Litters with soft tissue malformation, n (%)

0 (0.0)

0 (0.0)

1 (5.0)

1 (4.5)

 

7 (1.8)

Fetuses with soft tissue malformation, n (%)

0 (0.0)

0 (0.0)

2 (1.7)

1 (0.8)

 

13 (0.5)

F: fetal incidence; L: litter incidence, a: 2 fetuses from the same litter

b: not found in soft tissue malformations of HCD, but finding seen in one HCD fetus as external and skeletal malformations.

HCD: Historical Control Data (Sprague-Dawley rats from the Supplier Janvier; signed in December 2016): max fetal incidence (max litter incidence) for the findings; -: none in HCD.

Table 14: Fetal skeletal variations

 

Dose level (mg/kg/day)

0

50

250

1000

HCD

Number of litters

20

23

20

22

388

Number of fetuses

117

143

131

138

2596

Vertebrae

 

 

 

 

 

Cervical vertebra(e): incomplete ossification of centrum, F (L)

0.9 (5.0)

0.7 (4.3)

2.3 (15.0)

15.9a(50.0)b

18.3 (54.2)

Thoracic vertebra(e): incomplete ossification of centrum, F (L)

1.7 (5.0)

3.5 (21.7)

9.2c(30.0)

14.5a(45.5)b

29.6 (87.5)

Thoracic vertebra(e): bipartite ossification of centrum

0.9 (5.0)

 

3.1 (10.0)

3.6 (18.2)

5.0 (16.7)

Thoracic vertebrae: dumbbell

ossification of centrum

1.7 (10.0)

 

3.1 (10.0)

4.3 (18.2)

6.5 (30.0)

Caudal vertebrae: incomplete ossification of centrum

0.9 (5.0)

 

0.8 (5.0)

2.9 (9.1)

1.4 (10.0)

Metatarsal bone

 

 

 

 

 

Unossified 1stmetatarsal, F (L)

12.0 (45.0)

19.6 (69.6)

20.6 (55.0)

42.0a(86.4)b

36.8 (72.7)

Metatarsal: unossified

 

 

 

1.4 (4.5)d,e

5.4 (21.7)

Metacarpal bone

 

 

 

 

 

Incomplete ossification of metacarpal(s)

1.7 (10.0)

0.7 (4.3)

0.8 (5.0)

4.3 (22.7)

2.2 (10.0)

Unossified metacarpal(s)

0.9 (5.0)

 

1.5 (10.0)

3.6 (18.2)f

2.9 (15.0)

Head

 

 

 

 

 

Nasal: incomplete ossification

 

 

 

0.7 (4.5)d

0.8 (5.6)

Incisive bone: incomplete ossification

 

 

 

1.4 (4.5)d,e

0.7 (5.0)

Maxilla: incomplete ossification

 

 

 

1.4 (4.5)d,e

0.7 (5.0)

Forelimb

 

 

 

 

 

Radius: incomplete ossification

 

 

 

0.7 (4.5)e

-

Hindlimb

 

 

 

 

 

Tibia: incomplete ossification

 

 

 

0.7 (4.5)e

-

Fibula: incomplete ossification

 

 

 

0.7 (4.5)e

-

Litters with skeletal variation in total, n (%)

17 (85.0)

23 (100.0)

17 (85.0)

21 (95.5)

345 (88.9)

Fetuses with skeletal variation in total, n (%)

72 (61.5)

96 (67.1)

65 (49.6)

96 (69.6)

1267 (48.8)

F: fetal incidence; L: litter incidence.

a: p<0.001 for the number of affected fetuses or litters;b: p<0.01 for the number of affected fetuses or litters;c: p<0.05 for the number of affected fetuses or litters.

d and e: tow fetuses from the same litter; f: including these two fetuses

HCD: Historical Control Data (Sprague-Dawley rats from the Supplier Janvier; signed in December 2016): max fetal incidence (max litter incidence) for the findings.

Table 15: Fetal skeletal malformations

 

Dose level (mg/kg/day)

0

50

250

1000

HCD

Number of litters

20

23

20

22

388

Number of fetuses

117

143

131

138

2596

Nasal: absent

 

 

 

0.7 (4.5)a

-

Palate: split

 

 

 

1.4 (4.5)a, b

0.7 (5.0)

Fused sternebra(e)

0.9 (5.0)

 

 

 

1.3 (8.7)

Absent sternebra(e)

0.9 (5.0)

 

 

 

-

Forepaw: supernumerary digit(s)

 

 

 

0.7 (4.5)

-

Tibia: unossified (unilateral)

 

 

 

0.7 (4.5)a

-

Tibia: short

 

 

 

0.7 (4.5)b

-

Fibula: unossified (unilateral)

 

 

 

0.7 (4.5)a

-

Fibula: short

 

 

 

0.7 (4.5)b

-

Litters with skeletal malformation, n (%)

1 (5.0)

0 (0.0)

0 (0.0)

2 (9.1)

15 (3.9)

Fetuses with skeletal malformation, n (%)

1 (0.9)

0 (0.0)

0 (0.0)

3 (2.2)

18 (0.7)

F: fetal incidence; L: litter incidence.

a and b: fetuses from the same litter.

HCD: Historical Control Data (Sprague-Dawley rats from the Supplier Janvier; signed in December 2016): max fetal incidence (max litter incidence) for the findings; -: none in HCD

Conclusions:
The test item, Ethoxylated bisphenol A diacrylate, was administered by gavage, once daily, from Days 6 to 20 p.c. inclusive, to pregnant Sprague-Dawley rats at dosages of 50, 250 and 1000 mg/kg/day.

At 1000 mg/kg/day, lower fetal body weight (female fetuses) and increased fetal ossification delays were observed in presence of some signs of limited maternal toxicity (i.e. lower body weight gain and net body weight gain from Day 6 p.c.).
At 50 and 250 mg/kg/day, there were signs of slight fetal ossification delays.

Under the experimental conditions and results of this study:
¿ the No Observed Adverse Effect Level (NOAEL) for maternal parameters was considered to be 1000 mg/kg/day,
¿ the NOAEL for embryo-fetal development was considered to be 1000 mg/kg/day.
Executive summary:

The objective of this GLP study was to evaluate the potential toxic effects of the test item, on the pregnant female and on embryonic and fetal development, following daily oral administration (gavage) to pregnant female rats from implantation to the day before scheduled hysterectomy [Days 6 to 20 post-coitum (p.c.) inclusive].

 

Methods

Three groups of 24 time-mated female Sprague-Dawley rats received the test item at 50, 250 or 1000 mg/kg/day by oral route (gavage) once daily from Days 6 to 20 p.c. A constant dosage-volume of 5 mL/kg/day was used. Another group of 23 rats received the vehicle alone (corn oil) under the same experimental conditions, and acted as a control group.

 

Formulation concentrations were checked in the first day and last week of treatment in the study.

The animals were checked daily for mortality and clinical signs. Body weight and food consumption were recorded every 2 to 3 days. On Day 21 p.c., females were sacrificed and submitted to a macroscopic post-mortem examination of principal thoracic and abdominal organs. Hysterectomy was performed and the numbers of corpora lutea, implantations, uterine scars, early and late resorptions, live and dead fetuses were recorded. Placenta weight was recorded. The fetuses were sexed, weighed and examined for external, soft tissues and skeletal (cartilages + bones) abnormalities.

 

Results

The test item concentrations in the dose formulations analyzed were within an acceptable range of variations when compared with the nominal values (± 15% required). No test item was observed in the control dose formulations.

 

On Day 21 p.c., 20/23, 23/24, 20/24 and 22/24 females pregnant with live fetuses at 0, 50, 250 and 1000 mg/kg/day, respectively.

 

There were no deaths in the study and the only test item-related clinical sign was ptyalism noted in almost all females treated at 1000 mg/kg/day.

 

There were no test item-related effects on mean food consumption and mean body weight at any tested dose. The mean body weight gain from Days 9 to 12 p.c. was reduced in females treated at 1000 mg/kg/day (+15 g vs. +22 g in controls, p<0.001 vs. controls).

 

There were no test item-related necropsy findings in females.

At 1000 mg/kg/day, correlating with the effects on mean body weight change, there was a lower mean net body weight change from Day 6 p.c. (+33.9 g vs. +51.5 g p<0.01 vs. controls) but no toxicologically significant effects on mean carcass weight. There were no effects on mean carcass weight and on mean net body weight change from Day 6 p.c. at 50 and 250 mg/kg/day.

 

All above mentioned findings in dams were considered to be test item-related and non-adverse.

 

There were no test item treatment-related effects on mean hysterectomy data, mean sex ratio, mean placental weight and mean gravid uterus weight.

At 1000 mg/kg/day when compared with controls, female fetuses had a lower mean body weight (-6%, p<0.01), considered to be test item-related and non-adverse.

At fetal examination, there were no test item treatment-related external or soft tissue variations and malformations, and no test item treatment-related effects on cartilages.

At 1000 mg/kg/day, there were fetal skeletal ossification delays (test item-related and non-adverse) in cervical and thoracic vertebra centra, 1stmetatarsal bones and to a lesser extent in metacarpal bones and caudal vertebra centra. Similar trend but to a lesser extent was observed in some of these bones at 50 and/or 250 mg/kg/day. At 1000 mg/kg/day, a relationship between litter skeletal malformations and the test item treatment was considered unlikely.

 

Conclusion

The test item, Ethoxylated bisphenol A diacrylate, was administered by gavage, once daily, from Days 6 to 20 p.c. inclusive, to pregnant Sprague-Dawley rats at dosages of 50, 250 and 1000 mg/kg/day.

 

At 1000 mg/kg/day, lower fetal body weight (female fetuses) and increased fetal ossification delays were observed in presence of some signs of limited maternal toxicity (i.e.lower body weight gain and net body weight gain from Day 6p.c.).

At 50 and 250 mg/kg/day, there were signs of slight fetal ossification delays.

 

Under the experimental conditions and results of this study:

.            the No Observed Adverse Effect Level (NOAEL) for maternal parameters was considered to be 1000 mg/kg/day,

.            the NOAEL for embryo-fetal development was considered to be 1000 mg/kg/day.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
The study is considered to be reliable with a klimish score of 1.
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 study in rat (Bentz 2017) :

The objective of this GLP study was to evaluate the potential toxic effects of the test item, on the pregnant female and on embryonic and fetal development, following daily oral administration (gavage) to pregnant female rats from implantation to the day before scheduled hysterectomy [Days 6 to 20 post-coitum (p.c.) inclusive].

Three groups of 24 time-mated female Sprague-Dawley rats received the test item at 50, 250 or 1000 mg/kg/day by oral route (gavage) once daily from Days 6 to 20p.c. A constant dosage-volume of 5 mL/kg/day was used. Another group of 23 rats received the vehicle alone (corn oil) under the same experimental conditions, and acted as a control group.

On Day 21 p.c., 20/23, 23/24, 20/24 and 22/24 females pregnant with live fetuses at 0, 50, 250 and 1000 mg/kg/day, respectively.

There were no deaths in the study andthe only test item-related clinical sign was ptyalism noted in almost all females treated at 1000 mg/kg/day.

There were no test item-related effects on mean food consumption and mean body weight at any tested dose. The mean body weight gain from Days 9 to 12 p.c.was reduced in females treated at 1000 mg/kg/day (+15 g vs.+22 g in controls, p<0.001vs.controls).

There were no test item-related necropsy findings in females.

At 1000 mg/kg/day, correlating with the effects on mean body weight change, there was a lower mean net body weight change from Day 6p.c. (+33.9 gvs.+51.5 g p<0.01vs.controls) but no toxicologically significant effects on mean carcass weight. There were no effects on mean carcass weight and on mean net body weight change from Day 6p.c.at 50 and 250 mg/kg/day.

All above mentioned findings in dams were considered to be test item-related and non-adverse.

There were no test item treatment-related effects on mean hysterectomy data, mean sex ratio, mean placental weight andmean gravid uterus weight.

At 1000 mg/kg/day when compared with controls, female fetuses had a lower mean body weight (-6%, p<0.01), considered to be test item-related and non-adverse.

At fetal examination, there were no test item treatment-related external or soft tissue variations and malformations, and no test item treatment-related effects on cartilages.

At 1000 mg/kg/day, there were fetal skeletal ossification delays (test item-related and non-adverse) in cervical and thoracic vertebra centra, 1stmetatarsal bones and to a lesser extent in metacarpal bones and caudal vertebra centra. Similar trend but to a lesser extent was observed in some of these bones at 50 and/or 250 mg/kg/day. At 1000 mg/kg/day, a relationship between litter skeletal malformations and the test item treatment was considered unlikely.

 

At 1000 mg/kg/day, lower fetal body weight (female fetuses) and increased fetal ossification delays were observed in presence of some signs of limited maternal toxicity (i.e.lower body weight gain and net body weight gain from Day 6p.c.). At 50 and 250 mg/kg/day, there were signs of slight fetal ossification delays.

Under the experimental conditions and results of this study:

.            the No Observed Adverse Effect Level (NOAEL) for maternal parameters was considered to be 1000 mg/kg/day,

.            the NOAEL for embryo-fetal development was considered to be 1000 mg/kg/day.

Justification for classification or non-classification

Based on the results available on Ethoxylated bisphenol A diacrylate, no classification for Ethoxylated Bisphenol A diacrylate is required for reprotoxicity according to the Regulation EC n°1272/2008.

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