Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

Under the conditions of the study, the NOAEL for the F0 male and female systemic toxicity was 100 mg/kg/day. The NOAEL for F0 reproductive toxicity was >1000 mg/kg/day for both sexes. The NOAEL for F1 offspring toxicity was >1000 mg/kg/day. 

Link to relevant study records

Referenceopen allclose all

Endpoint:
one-generation reproductive toxicity
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
Deviations:
yes
Remarks:
- Due to the limited toxicity observed in previous studies for a similar test substance, only two BPA-DA dose groups were used
GLP compliance:
yes
Species:
rat
Strain:
other: CD® (Sprague-Dawley)
Sex:
male/female
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
Male and female CD (Sprague-Dawley) rats (the F0 generation) were administered BPA-DA orally by gavage at 0, 100, and 1000 mg/kg/day at a dose volume of 5 mL/kg/day in corn oil, ten/animals/sex/dose, for two weeks of prebreed exposure and two weeks of mating for F0 male and female parental animals. F0 females continued to be dosed for three weeks of gestation and through postnatal day (pnd) 3.
Details on mating procedure:
After the two-week prebreed exposure period, animals were randomly mated within treatment groups for a two-week mating period to produce the F1 generation.
Analytical verification of doses or concentrations:
yes
Duration of treatment / exposure:
Males = 4 weeks (2 weeks prebreed, 2 weeks mating)
Females = ~7 weeks (2 weeks prebreed, 2 weeks mating, 3 weeks gestation, and lactation through postnatal day 4)
Frequency of treatment:
Daily
Dose / conc.:
100 mg/kg bw/day (nominal)
Dose / conc.:
1 000 mg/kg bw/day (nominal)
No. of animals per sex per dose:
10/sex/group
Control animals:
yes, concurrent vehicle
Parental animals: Observations and examinations:
Body weights for the F0 males and females were recorded weekly during the prebreed and mating periods for both sexes and for F0 females during gestation. During lactation, F0 female body weights were obtained on pnd 0 and 4. Feed consumption was recorded weekly for the F0 males and females during the prebreed period, but not during the mating period. Feed consumption was recorded for the F0 females during gestation and through pnd 4 of lactation. Clinical signs were recorded at least once daily for all animals.
Litter observations:
On the day of birth (pnd 0), all live F1 pups were counted, sexed, weighed and examined as soon as possible. All stillborn pups or pups that died on the date of birth were sexed and counted. All pups were examined daily from birth through pnd 4 for survival and physical abnormalities.
Postmortem examinations (parental animals):
F0 males were sacrificed following the breeding period (after 28 days of dosing). F0 females with litters were sacrificed on pnd 4 and F0 females that did not produce a litter were sacrificed on gestation day (gd) 26 or 26 days after mating.
At the F0 parental animal necropsy, the following tissues were weighed and retained: testes, epididymides, prostate, seminal vesicles, ovaries, uterus. All gross lesions were also retained. Histopathology was performed on all retained reproductive tissues for the high dose and control males and females with special emphasis on stages of spermatogenesis in the male gonads and histopathology of interstitial testicular cell structure (10/sex/group). The uteri from the F0 females that failed to produce a litter by gd 26 or by 26 days post-mating were stained with potassium ferricyanide for confirmation of pregnancy.
Postmortem examinations (offspring):
Any pups dying during lactation were necropsied, if possible. On pnd 4, all live pups were examined sexed and weighted, then euthanized and discarded without further evaluation.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
- At 1000 mg/kg/day in parental males, treatment-related clinical observations were recorded for five males with audible respiration, two males with gasping, four males with sneezing, and one male with chromodacryorrhea. Other findings were not considered related to treatment.
- Treatment-related clinical observations at 1000 mg/kg/day in females included two to three females with audible respiration, sneezing and/or chromodacryorrhea. Other findings were not considered related to treatment.
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Description (incidence):
There were no treatment-related deaths for the F0 females or males.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
- For parental males at 1000 mg/kg/day, body weight was reduced on study days (sd) 21 and 28, by 7 and 5 % respectively. Body weight change was reduced at 1000 mg/kg/day for sd 14 to 21 and 0 to 28. At 100 mg/kg/day, there were no effects on body weight or body weight changes.
- There were no significant changes in F0 female body weights during the prebreed and mating periods; however, there was a decrease in body weight change from sd 7-14 at 1000 mg/kg/day. During gestation, there were no significant changes in body weight or body weight change for the F0 females. During lactation, there were no significant differences between groups in F0 maternal body weights or body weight change values.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
- At 1000 mg/kg/day in parental males, feed consumption (g/day) was increased from sd 7 to 14 and feed consumption (g/kg/day) was increased throughout the entire prebreed period. At 100 mg/kg/day, there were no effects on feed consumption.
- There were no significant changes in F0 female feed consumption during the prebreed and mating periods. During gestation, feed consumption (g/kg/day) was increased from gd 7 to 14 at 100 and 1000 mg/kg/day compared to controls. During lactation, there were no significant differences between groups in F0 maternal feed consumption values.
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):
effects observed, treatment-related
Description (incidence and severity):
Post-dose rooting is considered to be a behavioural response to taste aversion to the dosing formulations and not a toxic sign. Since there was a dose-related increase in the incidence of post-dose rooting (0, 2 and 3 females in the 0, 100 and 1000 mg/kg/day groups, respectively and 1, 2 and 5 males in the 0, 100 and 1000 mg/kg/day groups, respectively), it was presumed that the increasing concentrations of test material across groups caused the adverse taste reaction.
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
There were no treatment-related microscopic findings.
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):
During the post mating period, there were 9, 9, and 8 females in the 0, 100, and 1000 mg/kg/day dose groups, respectively, that were determined to be sperm positive; however, the total number of females confirmed pregnant at study completion was 9, 10 and 9, respectively. One pregnant female in the 100 mg/kg/day group was not identified as being sperm positive; one female each in the control and 1000 mg/kg/day groups was not pregnant, and one pregnant female at 1000 mg/kg/day did not deliver a litter. There was a statistically significant increase in precoital interval at 1000 /mg/kg/day although the increase was only approximately one day longer than the controls. There were no significant effects of exposure to the test material on F0 fertility, mating, pregnancy, preimplantation or postimplantation loss per litter, or the number of dead pups at birth.
Key result
Dose descriptor:
NOAEL
Remarks:
parental
Effect level:
ca. 100 mg/kg bw/day
Sex:
male/female
Basis for effect level:
other: see 'Remark'
Key result
Dose descriptor:
NOAEL
Remarks:
Reproduction
Effect level:
> 1 000 mg/kg bw/day
Sex:
male/female
Basis for effect level:
other: There were no significant effects of exposure to the test material on F0 fertility, mating, pregnancy, preimplantation or postimplantation loss per litter, or the number of dead pups at birth.
Clinical signs:
no effects observed
Dermal irritation (if dermal study):
not examined
Mortality / viability:
no mortality observed
Body weight and weight changes:
no effects observed
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:
not examined
Histopathological findings:
not examined
Other effects:
not examined
Behaviour (functional findings):
not examined
Developmental immunotoxicity:
not examined
There was no evidence of F1 offspring toxicity at any dose.
Key result
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
> 1 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: There was no evidence of F1 offspring toxicity at any dose.
Reproductive effects observed:
no

Summary of F0 Adult Systemic Toxicity-Key Parameters and Statistically Significant Differences

Bisphenol A Dianhydride (mg/kg/day)

0

100

1000

F0 MALES

Deaths

0/10

0/10

0/10

Body Weights

sd 21

---

---

<

sd 2

---

---

<

Weight Change

sd 14-21

---

---

<

sd 0-28

---

---

<

Feed Consumption:      

g/day

sd 7-14

---

---

<<

                g/kg/day

sd 0-7

---

---

<<

sd 7-14

---

---

<<<

sd 0-14

---

---

<<<

Necropsy

Final Body Weight at Necropsy

---

---

<<<

Organ Weights

---

---

---

F0 FEMALES

Deaths

0/10

0/10

0/10

Prebreed, Mating, and Postmating (sd 0-42) Exposure

Body Weights

---

---

---

Weight Change

sd 7-14

---

---

<

Feed Consumption:       g/day

---

---

---

                                                g/kg/day

---

---

---

Gestation

Body Weights

---

---

---

Weight Change

---

---

---

Feed Consumption:      

g/day

---

---

---

                g/kg/day

gd 7-14

---

>>

>

Lactation (pnd 0-21)

Body Weights

---

---

---

Weight Change

---

---

---

Feed Consumption:       g/day

---

---

---

                                                g/kg/day

---

---

---

Necropsy

Final Body Weight at Necropsy

---

---

---

Organ Weights

Paired Ovary Weight

---

---

<<

Relative Paired Ovary Weight

---

---

<

>, >>, >>> = statistically significant increase; p = 0.05, p = 0.01 and p = 0.001, respectively

<, <<, <<< = statistically significant decrease; p = 0.05, p = 0.01 and  p = 0.001, respectively

--- = no statistically significant difference

Summary of F0 Parental Male and Female Reproductive Toxicity

F0

Bisphenol A Dianhydride (mg/kg/day)

0

100

1000

F0 Females

No. Females on Study

10

10

10

No. Females Paired

10

10

10

No. Females that Mated

10

10

9

Mating Index (# females mated/# females paired)

100.0

100.0

90.0

No. Pregnant Females

9

10

9

Fertility Index (# pregnant females/# females that mated)

90.0

100.0

100.0

No. of Females with Live Litters (pnd 0)

9

10

8a

Gestational Index (# females with live litters/# females pregnant)

100.0

100.0

88.9

F0 Males

No. Males on Study

10

10

10

No. Males Paired

10

10

10

No. Males that Mated

10

10

9

Mating Index (# males mated/# males paired)

100.0

100.0

90.0

No. Males Siring Litters

9

10

9

Fertility Index (# males siring litters/# males that mated)

90.0

100.0

100.0

Pregnancy Index (# females with live litters/# males that mated)

90.0

100.0

88.9

Precoital Interval (days)

1.8 ± 0.2

2.1 ± 0.4

3.1 ± 0.5*

Gestational Length (days)

22.0 ± 0.0

22.3 ± 0.2

22.3 ± 0.2

No. Live Litters

Postnatal Day 0

9

10

8

Postnatal Day 4

9

10

8

No. Corpora Lutea per Dam

15.33 ± 1.25

14.30 ± 1.32

14.10 ± 1.51

% Preimplantation Loss per Litter

5.20 ± 2.45

8.69 ± 3.88

6.33 ± 2.91b

Average No.  Implantation Sites per Litter

15.89 ± 1.12

14.20 ± 1.55

16.00 ± 1.00

% Postimplantation Loss per Litter

5.77 ± 1.69

14.70 ± 5.52

19.82 ± 10.37

Average No. of Live Pups on Postnatal Day 0

14.8 ± 1.2

12.7 ± 1.7

13.4 ± 1.1

Average No. of Dead Pups on Postnatal Day 0

0.2 ± 0.1

0.0 ± 0.0

0.6 ± 0.3

Average Total Number of Pups on Postnatal Day 0

15.0 ± 1.1

12.7 ± 1.7

14.0 ± 1.0

Stillbirth Index (# dead on pnd 0/total # on pnd 0)

1.9 ± 1.3

0.0 ± 0.0

5.0 ± 2.9

Live Birth Index (# live on pnd 0/total# on pnd 0)

98.1 ± 1.3

100.0 ± 0.0

95.0 ± 2.9

4 Day Survival Index (# surviving 4 days/# live on pnd 0)

98.8 ± 0.8

92.8 ± 4.9

98.2 ± 1.8

a  A female was pregnant (20 implantation sites at necropsy) but did not deliver a litter.

b One female had corpora lutea, but no implantation sites; therefore, preimplantation loss could not be calculated.

Summary of F1 Offspring Toxicity

F1

Bisphenol A Dianhydride (mg/kg/day)

0

10

100

No. Live Litters

Postnatal Day 0

9

10

8

Postnatal Day 4

9

10

8

Average No. of Live Pups per Litter (pnd 0)

14.8 ± 1.2

12.7 ± 1.7

13.4 ± 1.1

Average No. of Live Pups per Litter (pnd 4)

14.6 ± 1.1

12.3 ± 1.7

13.3 ± 1.2

Average Pup Body Weight (g) per Litter (pnd 0)

6.33 ± 0.14

7.03 ± 0.17**

6.40 ± 0.14

Average Male Body Weight (g) per Litter (pnd 0)

6.45 ± 0.17

7.16 ± 0.19*

6.49 ± 0.16

Average Female Body Weight (g) per Litter (pnd 0)

6.22 ± 0.13

6.91 ± 0.17**

6.30 ± 0.13

Average Pup Body Weight (g) per Litter (pnd 4)

9.96 ± 0.33

11.22 ± 0.52

10.81 ± 0.51

Average Male Body Weight (g) per Litter (pnd 4)

10.20 ± 0.36

11.44 ± 0.58

11.02 ± 0.52

Average Female Body Weight (g) per Litter (pnd 4)

9.71 ± 0.30

10.93 ± 0.54

10.63 ± 0.52

% Percent Male Pups per Litter (pnd 0)

54.9 ± 5.1

48.5 ± 3.5

44.4 ± 3.9

% Percent Male Pups per Litter (pnd 4)

55.0 ± 5.2

60.2 ± 6.5

48.8 ± 2.7

* p = 0.05; ** p = 0.01

Conclusions:
Under the conditions of this study, the NOAEL for the F0 male and female systemic toxicity was 100 mg/kg/day. The NOAEL for F0 reproductive toxicity was >1000 mg/kg/day for both sexes. The NOAEL for F1 offspring toxicity was >1000 mg/kg/day.
Executive summary:

A screening reproduction/development test was conducted using methods comparable to OECD guideline 421 under GLP conditions (RTI International, 2005).

Sprague-Dawley male and female rats were exposed to the test material via gavage in corn oil at dose levels of 0, 100, and 1000 mg/kg/day (10/sex/group). Due to the limited toxicity observed in previous studies for a similar test substance, only two dose groups were used.

Males were dosed for 4 weeks (2 weeks prior to breeding, 2 week mating period) and females were dosed for ~7 weeks (2 weeks prior to breeding, 2 week mating period, 3 week gestation period and lactation through postnatal day 4).

Minimal systemic toxicity was present in males and females through the course of the study at 1000 mg/kg/day. In the F0 males, the only adverse effects were respiratory signs, a 5 to 7 % reduction in body weights, and reductions in body weight changes. In the F0 females, the only adverse effects were decreases in high dose body weight change (Days 7 to 14) and treatment related clinical signs at the high dose. At the high dose there was an increase in the pre-coital interval, but no effect on fertility. There was no evidence of reproductive toxicity in the F0 females at any dose, or any toxicity in the F1 offspring. At necropsy, for the F0 males and females, there were no treatment effects with the exception of decreases in the absolute and relative paired ovary weights. 

Under the conditions of this study the NOAEL for the F0 male and female systemic toxicity was 100 mg/kg/day. The NOAEL for F0 reproductive toxicity was >1000 mg/kg/day for both sexes. The NOAEL for F1 offspring toxicity was >1000 mg/kg/day. 

Endpoint:
extended one-generation reproductive toxicity - basic test design (Cohorts 1A, and 1B without extension)
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
other:
Reproductive effects observed:
not specified
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:
The study was conducted in accordance with a standardised guideline and under GLP conditions. The quality of the database is therefore considered to be good.
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

A screening reproduction/development test was conducted using methods comparable to OECD guideline 421 under GLP conditions (RTI International, 2005). The study was awarded a reliability score of 1 in accordance with the criteria set forth by Klimisch et al. (1997).

Sprague-Dawley male and female rats were exposed to the test material via gavage in corn oil at dose levels of 0, 100, and 1000 mg/kg/day (10/sex/group). Due to the limited toxicity observed in previous studies for a similar test substance, only two BPA-DA dose groups were used.

Males were dosed for 4 weeks (2 weeks prior to breeding, 2 week mating period) and females were dosed for ~7 weeks (2 weeks prior to breeding, 2 week mating period, 3 week gestation period and lactation through postnatal day 4).

Minimal systemic toxicity was present in males and females through the course of the study at 1000 mg/kg/day. In the F0 males, the only adverse effects were respiratory signs, a 5 to 7 % reduction in body weights, and reductions in body weight changes. In the F0 females, the only adverse effects were decreases in high dose body weight change (Days 7 to 14) and treatment related clinical signs at the high dose. At the high dose there was an increase in the pre-coital interval, but no effect on fertility. There was no evidence of reproductive toxicity in the F0 females at any dose, or any toxicity in the F1 offspring. At necropsy, for the F0 males and females, there were no treatment effects with the exception of decreases in the absolute and relative paired ovary weights. Based on these results, the NOAEL for the F0 male and female systemic toxicity was 100 mg/kg/day. The NOAEL for F0 reproductive toxicity was > 1000 mg/kg/day for both sexes. The NOAEL for F1 offspring toxicity was > 1000 mg/kg/day. 

In accordance with Column 2 of REACH Annex IX, Section 8.7, further reproductive toxicity studies should be proposed if the available repeated dose toxicity studies indicate adverse effects on reproductive organs or tissues or reveal other concerns in relation with reproductive toxicity.

Moreover, in accordance with Column 2 of REACH Annex IX, Section 8.7.3, the Extended One-Generation Reproductive Toxicity Study shall be proposed if the substance has uses leading to significant exposure of consumers or professionals and if: (1) the substance displays genotoxic effects in somatic cell mutagenicity tests in vivo which could lead to classifying it as Mutagen Category 2, or (2) there are indications that the internal dose for the substance and/or any of its metabolites will reach a steady state in the test animals only after an extended exposure, or (3) there are indications of one or more relevant modes of action related to endocrine disruption from available in vivo studies or non-animal approaches. Professional and consumer use are not relevant to the substance, and there is no consumer exposure from articles. Additionally, the substance is not classified as a mutagen, there are no indications that the internal dose for the substance and/or any of its metabolites will reach a steady state in the test animals only after an extended exposure, and there are no indications of modes or action related to endocrine disruption in available in vivo studies.

It is therefore considered to be justified to omit this study.

Effects on developmental toxicity

Description of key information

In a screening reproduction/development test conducted in the rat using methods comparable to OECD guideline 421, the NOAEL for F0 reproductive toxicity was >1000 mg/kg/day. The NOAEL for F1 offspring toxicity was >1000 mg/kg/day.

In a developmental test conducted in the rabbit using methods comparable to OECD 414 and EPA OPPTS 870.3700, the NOAEL for developmental toxicity was considered to be ≥1000 mg/kg bw.

In a developmental test conducted in the rat using methods comparable to OECD 414 and EPA OPPTS 870.3700, the NOAEL for developmental toxicity was considered to be ≥1000 mg/kg bw.

Link to relevant study records

Referenceopen allclose all

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
yes
Remarks:
insufficient number of pregnant dams/dose group, dosing from days 6-18 of gestation
Qualifier:
equivalent or similar to
Guideline:
EPA OPPTS 870.3700 (Prenatal Developmental Toxicity Study)
Deviations:
yes
Remarks:
insufficient number of pregnant dams/dose group, dosing from days 6-18 of gestation
GLP compliance:
yes
Limit test:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
Ninety mature New Zealand White female rabbits were obtained from Dutchland Laboratory Animals, Inc., Denver, PA for use in this study. The animals were acclimated for a minimum of 22 days prior to the initiation of the study. During the period of acclimation, the rabbits were examined for general health and appearance. The animals were uniquely identified by ear tag and provided commercial rabbit ration (Purina Lab Rabbit Chow) and tap water ad libitum. The environment of the study room was maintained at 70-78 °F, relative humidity of 53-86% and a 12-hour light/dark cycle.
Route of administration:
oral: gavage
Vehicle:
CMC (carboxymethyl cellulose)
Details on exposure:
Five groups were included in this study; for the purposes of this summary, only three groups (control, positive control, and BPA-DA treated) will be discussed. Sixteen animals per group (to obtain at least 12 pregnant) were treated with vehicle (0.5% carboxymethyl cellulose), positive control (thalidomide; 150 mg/kg/day) or BPA-DA (1000 mg/kg/day). Thalidomide and BPA-DA were suspended in vehicle to provide dose volumes of 1.5 and 2.5 mL/kg, respectively. Control dose volume was 4.0 mL/kg.
Analytical verification of doses or concentrations:
not specified
Details on mating procedure:
At Day 0 of gestation, the body weights ranged from 2845 to 4700 grams. The animals were artificially inseminated with sperm from the laboratory breeding stock five hours after induction of ovulation with chorionic gonadotropin.
Duration of treatment / exposure:
Days 6 through 18 of gestation. ( The dose was administered from gestation day (gd) 6 through 18, approximately the same time each day, and was based on each individual body weight on gd 6 (starting on gd 11, two animals in the control group, four animals in the thalidomide group and three animals in the BPA-DA group were dosed based on gd 11 body weight).)
Frequency of treatment:
Daily
Duration of test:
29 Days
Dose / conc.:
1 000 mg/kg bw/day (nominal)
No. of animals per sex per dose:
16/group
Control animals:
yes, concurrent vehicle
other: Positive Control (Thalidomide)
Maternal examinations:
All of the animals were observed daily for mortality, moribundity and clinical signs. Body weights were recorded on gd 0, 6, 11, 15, 19 and 29. Individual food consumption was recorded weekly.
Ovaries and uterine content:
On gd 29, the animals were sacrificed, examined for gross pathology of the external surface and viscera, and the uterus excised and weighed. The fetuses were taken by cesarean section and the following recorded for each litter: the number of corpora lutea per ovary; the number and placement of uterine implantation sites; live and dead fetuses; early and late resorptions; and any other abnormalities.
Cesarean sections were also performed on dams that were found dead, sacrificed moribund or sacrificed due to early delivery. The number of corpora lutea, implantations, resorptions and live or dead fetuses was recorded.
Fetal examinations:
Fetuses were removed from the placenta, individually identified, examined externally, weighed and measured from the frontal-parietal suture to the base of the tail (crown-rump distance).
The unfixed fetuses underwent visceral examination according to the method of Staples. All of the fetuses were opened by longitudinal incision, the sex determined and examined grossly both externally and internally. Major organs were inspected in situ with special attention to the heart and major blood vessels. The heads of approximately one-third of the fetuses were removed, fixed in Bouin’s solution, sectioned by Wilson’s freehand sectioning technique for examination of the eyes, palate, nasal septum and brain. The prepared sections were then re-examined against a light box with the aid of magnification.
Following visceral examination, all fetuses (minus the head for approximately one-third of the fetuses) were eviscerated and placed in 95% ethyl alcohol. After fixation and dehydration, the skeletons were stained in a potassium hydroxide-alizarin red solution. The skull, vertebral column, rib cage, pectoral and pelvic girdles, long bones and extremities of each skeleton were examined for degree of ossification, bone alignment, and possible anomalies. Examinations were performed with the aid of magnification on a light box.
Statistics:
Mean maternal body weight changes, food consumption, percentage data (implantations, resorptions and males), and fetal viability were analysed in the following order: Levene’s test for homogeneity of variance; if the variances proved to be homogeneous, the data were analysed by one-way classification analysis of variance (ANOVA); if the variance proved to be heterogeneous, a series of transformations was performed until homogeneity was achieved followed by ANOVA. If ANOVA was significant, the Games and Howell modification of the Tukey-Kramer honestly significant difference test was used to compare groups. Pregnancy rates were analysed by Fisher’s exact test. External, visceral, and skeletal anomalies were evaluated by a multiple proportions test. Analysis of covariance (ANCOVA) was used to analyse mean fetal weights and lengths with the litter used as the experimental unit. Levene’s test and ANOVA were evaluated at the 5% one-tailed probability level. Control vs. treatment group mean comparisons were evaluated at the 5% two-tailed probability level.
Clinical signs:
no effects observed
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Weight loss was observed in the test material treated groups during the treatment period. Statistical evaluation of body weight change did not, however, reveal any significant differences between treated and control groups.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
No effects on food consumption were observed.
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:
no effects observed
Description (incidence and severity):
No effects in the gross pathology of the dams were observed.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Other effects:
not examined
Details on maternal toxic effects:
Maternal toxic effects: yes

Details on maternal toxic effects:
Weight loss was observed in the thalidomide- and BPA-DA-treated groups during the treatment period. Statistical evaluation of body weight change did not, however, reveal any significant differences between treated and control groups. No effects on food consumption or gross pathology of the dams were observed.
There were no differences from control in the thalidomide or BPA DA dose groups for maternal, ovarian or uterine data. The thalidomide treated group exhibited changes consistent with the known teratogenic effect of this compound.
Key result
Dose descriptor:
NOAEL
Effect level:
> 1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
body weight and weight gain
Fetal body weight changes:
no effects observed
Reduction in number of live offspring:
no effects observed
Changes in sex ratio:
no effects observed
Changes in litter size and weights:
no effects observed
Changes in postnatal survival:
no effects observed
External malformations:
no effects observed
Skeletal malformations:
no effects observed
Visceral malformations:
no effects observed
Other effects:
not examined
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects: no effects

Details on embryotoxic / teratogenic effects: There were no effects on any fetal parameters from BPA-DA treatment.
Key result
Dose descriptor:
NOEL
Effect level:
> 1 000 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
other: teratogenicity
Abnormalities:
no effects observed
Developmental effects observed:
no

Summary of Mean Ovarian, Uterine, and Litter Data

Parameter

Control

(Vehicle)

Thalidomide (Positive Control)

BPA-DA

(1000 mg/kg/day)

Number of dams

16

16

16

Number pregnant

14

16

13

Pregnancy rate (%)

88

100

81

Number dams surviving to gd 29

13

15*

12

    (survival  rate)

93%

100%

92%

Mean number of

   Corpora lutea

13.4

12.2

11.4

   Implantations

9.4

8.3

6.5

   Resorptions-total

1.2

5.3

2.1

   Fetuses - live

7.5

3.4

4.5

                - dead

0.5

0

0.1

Indices (mean per litter)

   Implantation  efficiency (%)

73.6

68.1

57

   Incidence of resorption (%)

17.2

61

31

   Incidence of fetal mortality (%)

3.8

0

0.9

   Incidence of fetal viability (%)

79.2

39.1

68.1

Live fetuses

   Mean body weight (g) - males

40.91

38.36

42.8

                                       - females

39.9

37.6

41.78

   Mean length (cm)        - males

9.49

9.03

9.42

                                       - females

9.33

8.92

9.39

   Percent Males

51.5

58

47.1

   Mean uterine weights - gravid (g)

485.3

228.3

315.4

* One animal died accidentally on gd 8

Summary of Mean Incidence of Abnormal Fetuses per Litter

Parameter

Control

(Vehicle)

Thalidomide (Positive Control)

BPA-DA

(1000 mg/kg/day)

External

  # of litters examined

12

11

9

  # of litters with anomalous fetuses

2

10*

3

  % of litters with anomalous fetuses

16.7

90.9

33.3

 

  Mean values (per litter)

    # of fetuses with variants

0

0.5

0

    Incidence of variants (%)

0

14.4

0

    # of fetuses with anomalies

0.3

2.7

0.6

    Incidence of anomalies (%)

2.4

64.1

8.2

Visceral - Fetal Heads

  # of litters examined

12

9

8

  # of litters with anomalous fetuses

0

3

0

  % of litters with anomalous fetuses

0

33.3

0

 

  Mean values (per litter)

    # of fetuses with variants

0

0.1

0

    Incidence of variants (%)

0

3.7

0

    # of fetuses with anomalies

0

0.4

0

    Incidence of anomalies (%)

0

16.7

0

Visceral - Torso and Limbs

  # of litters examined

12

11

9

  # of litters with anomalous fetuses

0

8*

1

  % of litters with anomalous fetuses

0

72.7

11.1

 

  Mean values (per litter)

    # of fetuses with variants

0.8

2.9

0.4

    Incidence of variants (%)

11.1

63.9

8.7

    # of fetuses with anomalies

0

1.5

0.1

    Incidence of anomalies (%)

0

38.8

1.6

Skeletal - Skulls

  # of litters examined

12

11

9

  # of litters with anomalous fetuses

0

2

0

  % of litters with anomalous fetuses

0

18.2

0

 

  Mean values (per litter)

    # of fetuses with variants

0.5

1.6

1.2

    Incidence of variants (%)

11.3

60.9

33.7

    # of fetuses with anomalies

0

0.2

0

    Incidence of anomalies (%)

0

11.4

0

Skeletal - Torso and Limbs

  # of litters examined

12

11

9

  # of litters with anomalous fetuses

0

10*

1

  % of litters with anomalous fetuses

0

90.9

11.1

 

  Mean values (per litter)

    # of fetuses with variants

0.6

3.9

0.8

    Incidence of variants (%)

6.9

91.7

16.4

    # of fetuses with anomalies

0

2.2

0.1

    Incidence of anomalies (%)

0

54.4

1.2

* Statistically significantly different from vehicle control group (p < 0.05)

Conclusions:
Based on the results of this study, the test material is not a developmental toxin.
Executive summary:

The developmental toxicity potential of BPA-DA in the rabbit was investigated in a study conducted using methodology equivalent to OECD 414 and EPA OPPTS 870.3700 under GLP conditions (Hazleton Laboratories America, Inc., 1983).

Sixteen New Zealand White rabbits were dosed with the test material at 1000 mg/kg bw by gavage in CMC (carboxymethyl cellulose) on Days 6 through 18 of gestation. A further 16 were dosed with the vehicle alone.

A reduction in maternal bodyweight was seen at 1000 mg/kg, however, it was not statistically significant. There were no effects on any foetal parameters from test material treatment and the NOAEL for developmental toxicity was considered to be ≥1000 mg/kg bw.

Based on the results of this study, the test material is not a developmental toxin.

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study
Qualifier:
equivalent or similar to
Guideline:
EPA OPPTS 870.3700 (Prenatal Developmental Toxicity Study)
Deviations:
yes
Remarks:
insufficient number of pregnant dams/dose group, dosing from days 6-18 of gestation
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
yes
Remarks:
insufficient number of pregnant dams/dose group, dosing from days 6-18 of gestation
GLP compliance:
yes
Limit test:
yes
Species:
rat
Strain:
other: Crl:CD®(SD)BR
Details on test animals and environmental conditions:
One hundred twenty, successfully mated Sprague-Dawley female rats, obtained from Charles River Breeding Laboratories, Inc., (Portage, MI), were used in this study. Prior to in-house breeding, the rats were examined for general health and appearance. The animals were uniquely identified by ear tag and provided commercial rat ration (Purina Certified Rodent Chow) and tap water ad libitum. The environment of the study room was monitored daily and a 12-hour light/dark cycle was used.
Route of administration:
oral: gavage
Vehicle:
CMC (carboxymethyl cellulose)
Details on exposure:
Twenty-four animals per group were treated with vehicle (0.5% carboxymethyl cellulose) or BPA-DA (1000 mg/kg/day). Dose volume was 10.0 mL/kg. The dose was administered from gestation day (gd) 6 through 15, approximately the same time each day, and was based on the most recently recorded body weight.
Analytical verification of doses or concentrations:
not specified
Details on mating procedure:
Animals were mated one male to one female for 17 days. The day that vaginal sperm or a copulation plug was observed was designated Day 0 of gestation. At Day 0 of gestation, the body weights ranged from 200 to 289 g. Six groups were included in this study; for the purposes of this summary, only two groups (control, and BPA-DA-treated) will be discussed.
Duration of treatment / exposure:
The dose was administered from gestation day (gd) 6 through 15, approximately the same time each day, and was based on the most recently recorded body weight.
Frequency of treatment:
daily
Duration of test:
20 days
Dose / conc.:
1 000 mg/kg bw/day (nominal)
No. of animals per sex per dose:
24/group
Control animals:
yes, concurrent vehicle
Maternal examinations:
All of the animals were observed daily for mortality, moribundity and clinical signs. Body weights and food consumption were recorded on gd 0, 6, 8, 12, 16 and 20.
Ovaries and uterine content:
On gd 20, the animals were sacrificed, examined for gross pathology of the external surface and viscera, and the uterus excised and weighed.
Fetal examinations:
The fetuses were taken by cesarean section and the following recorded for each litter: the number of corpora lutea per ovary; the number and placement of uterine implantation sites; live and dead fetuses; early and late resorptions; and any other abnormalities. Fetuses were removed from the placenta, individually identified, examined externally, and weighed.
Visceral Examination of Fetuses: Approximately one-third of the live fetuses were selected for visceral examination according to the method of Wilson.
Skeletal Examination of Fetuses: The remaining fetuses were eviscerated and placed in 95% ethyl alcohol. After fixation and dehydration, the skeletons were stained in a potassium hydroxide-alizarin red solution. The skull, vertebral column, rib cage, pectoral and pelvic girdles, long bones and extremities of each skeleton were examined for degree of ossification, bone alignment, and possible anomalies.
Statistics:
Mean maternal body weight changes, food consumption, percentage data (implantations, resorptions and males), and fetal viability were analysed in the following order: Levene’s test for homogeneity of variance; if the variances proved to be homogeneous, the data were analysed by one-way classification analysis of variance (ANOVA); if the variance proved to be heterogeneous, a series of transformations was performed until homogeneity was achieved followed by ANOVA. If ANOVA was significant, the Dunnett’s test was used to compare groups. Pregnancy rates, clinical observations and fetal skeletal observations were analysed by Cochran-Armitage and Fisher-Irwin Exact Tests. Analysis of covariance (ANCOVA) was used to analyse mean fetal weights with the litter used as the experimental unit. Levene’s test and ANOVA were evaluated at the 5% one-tailed probability level. Control vs. treatment group mean comparisons were evaluated at the 5% two-tailed probability level.
Clinical signs:
no effects observed
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Mean maternal body weight was significantly lower than the control group on gd 8, 12, 16 and 20 and mean weight gain was significantly lower than control for gd 6-16.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Mean maternal food consumption was significantly lower than control for gd 6-8 and 8-12.
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:
no effects observed
Gross pathological findings:
no effects observed
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
not examined
Other effects:
not examined
Number of abortions:
no effects observed
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
no effects observed
Early or late resorptions:
no effects observed
Dead fetuses:
no effects observed
Changes in pregnancy duration:
no effects observed
Changes in number of pregnant:
no effects observed
Other effects:
not examined
Details on maternal toxic effects:
There were no differences from control in the BPA-DA dose groups for ovarian or uterine data.
Key result
Dose descriptor:
LOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Basis for effect level:
other: maternal toxicity
Fetal body weight changes:
no effects observed
Reduction in number of live offspring:
no effects observed
Changes in sex ratio:
no effects observed
Changes in litter size and weights:
no effects observed
Changes in postnatal survival:
no effects observed
External malformations:
no effects observed
Skeletal malformations:
no effects observed
Visceral malformations:
no effects observed
Other effects:
not examined
Details on embryotoxic / teratogenic effects:
There were no treatment-related effects on any fetal parameters from BPA-DA treatment.
Key result
Dose descriptor:
NOAEL
Effect level:
>= 1 000 mg/kg bw/day (nominal)
Basis for effect level:
other: teratogenicity
Abnormalities:
no effects observed
Key result
Developmental effects observed:
no

Summary of Mean Ovarian, Uterine, and Litter Data

Parameter

Control

(Vehicle)

BPA-DA

(1000 mg/kg/day)

Number of dams

24

24

Number pregnant

23

24

Pregnancy rate (%)

96

100

Number dams surviving to gd 29

24

24

    (survival  rate)

100%

100%

Mean number of

   Corpora lutea

16.9

15.9

   Implantations (% Efficiency)

14.8 (89)

14.3 (90)

   Resorptions-total

0.9

0.4

   Fetuses - live

14

13.8

                - dead

0

0

Live fetuses

   Mean body weight (g) - males

3.6

3.5

                                      - females

3.3

3.4

   Mean uterine weights - gravid (g)

76.3

75.2

Summary of Mean Incidence of Abnormal Fetuses per Litter

Parameter

Control

(Vehicle)

BPA-DA

(1000 mg/kg/day)

External Variations

  Litter Incidence

    # of litters examined

23

24

    # of litters with anomalous fetuses

3

3

    % of litters with anomalous fetuses

13

13

 

  Fetal Incidence

    # of fetuses with variants

4

4

    Incidence of variant (%)

1.2

1.2

 

External Malformations

  Litter Incidence

    # of litters examined

23

24

    # of litters with anomalous fetuses

1

0

    % of litters with anomalous fetuses

4.3

0

 

  Fetal Incidence

    # of fetuses with variants

1

0

    Incidence of variant (%)

0.3

0

 

Soft Tissue Variations

  Litter Incidence

    # of litters examined

23

24

    # of litters with anomalous fetuses

6

6

    % of litters with anomalous fetuses

26

25

 

  Fetal Incidence

    # of fetuses with variants

9

9

    Incidence of variant (%)

9.2

8.8

 

Soft Tissue Malformations

  Litter Incidence

    # of litters examined

23

24

    # of litters with anomalous fetuses

0

0

    % of litters with anomalous fetuses

0

0

 

  Fetal Incidence

    # of fetuses with variants

0

0

    Incidence of variant (%)

0

0

 

Skeletal Variations

  Litter Incidence

    # of litters examined

23

24

    # of litters with anomalous fetuses

23

23

  % of litters with anomalous fetuses

100

96

 

  Fetal Incidence

    # of fetuses with variants

120

120

    Incidence of variant (%)

54

52

 

Skeletal Malformations

  Litter Incidence

    # of litters examined

23

24

    # of litters with anomalous fetuses

1

0

    % of litters with anomalous fetuses

4.3

0

 

  Fetal Incidence

    # of fetuses with variants

1

0

    Incidence of variant (%)

0.4

0

Conclusions:
Under the conditions of this study, the test material is not a developmental toxin.
Executive summary:

The developmental toxicity potential of the test material in the rat was investigated in a study conducted using methodology equivalent to OECD 414 and EPA OPPTS 870.3700 under GLP conditions (Hazleton Laboratories America, Inc., 1987).

Twenty-four Sprague-Dawley rats were dosed with the test material at 1000 mg/kg bw by gavage in CMC (carboxymethyl cellulose) on Days 6 through 15 of gestation. A further 24 were dosed with the vehicle alone.

A reduction in maternal bodyweight was seen at 1000 mg/kg. There were no effects on any foetal parameters from test material treatment and the NOAEL for developmental toxicity was considered to be ≥1000 mg/kg bw.

Based on the results of this study, the test material is not a developmental toxin.

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
other: OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
Deviations:
yes
Remarks:
Due to the limited toxicity observed in previous studies for a similar test substance, only two BPA-DA dose groups were used
GLP compliance:
yes
Species:
rat
Strain:
other: CD® (Sprague-Dawley)
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
CD (Sprague-Dawley) rats (the F0 generation) were administered BPA-DA orally by gavage at 0, 100, and 1000 mg/kg/day at a dose volume of 5 mL/kg/day in corn oil, ten/animals/sex/dose, for two weeks of pre-breed exposure and two weeks of mating followed by three weeks of gestation and through postnatal day (pnd) 3.
Analytical verification of doses or concentrations:
yes
Details on mating procedure:
After the two-week prebreed exposure period, animals were randomly mated within treatment groups for a two-week mating period to produce the F1 generation.
Duration of treatment / exposure:
~7 weeks (2 weeks prebreed, 2 weeks mating, 3 weeks gestation, and lactation through postnatal day 4)
Frequency of treatment:
Daily
Dose / conc.:
100 mg/kg bw/day (nominal)
Dose / conc.:
1 000 mg/kg bw/day (nominal)
No. of animals per sex per dose:
10/sex/group
Control animals:
yes, concurrent vehicle
Maternal examinations:
Body weights were recorded weekly during the prebreed, mating and gestation periods. During lactation, F0 female body weights were obtained on pnd 0 and 4. Feed consumption was recorded weekly during the prebreed period, but not during the mating period. Feed consumption was recorded for the F0 females during gestation and through pnd 4 of lactation. Clinical signs were recorded at least once daily for all animals.

F0 females with litters were sacrificed on pnd 4 and F0 females that did not produce a litter were sacrificed on gestation day (gd) 26 or 26 days after mating.
At the F0 parental animal necropsy, the following tissues were weighed and retained: ovaries and uterus. All gross lesions were also retained. Histopathology was performed on all retained reproductive tissues for the high dose and control animals. The uteri from the F0 females that failed to produce a litter by gd 26 or by 26 days post-mating were stained with potassium ferricyanide for confirmation of pregnancy.
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Fetal examinations:
On the day of birth (pnd 0), all live F1 pups were counted, sexed, weighed and examined as soon as possible. All stillborn pups or pups that died on the date of birth were sexed and counted. All pups were examined daily from birth through pnd 4 for survival and physical abnormalities.

Any pups dying during lactation were necropsied, if possible. On pnd 4, all live pups were examined sexed and weighed, then euthanised and discarded without further evaluation.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Treatment-related clinical observations at 1000 mg/kg/day included two to three females with audible respiration, sneezing and/or chromodacryorrhea. Other findings were not considered related to treatment.
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Description (incidence):
There were no treatment-related deaths for the F0 females.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
There were no significant changes in F0 female body weights during the prebreed and mating periods; however, there was a decrease in body weight change from sd 7-14 at 1000 mg/kg/day. During gestation, there were no significant changes in body weight or body weight change for the F0 females. During lactation, there were no significant differences between groups in F0 maternal body weights or body weight change values.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
There were no significant changes in F0 female feed consumption during the prebreed and mating periods. During gestation, feed consumption (g/kg/day) was increased from gd 7 to 14 at 100 and 1000 mg/kg/day compared to controls. During lactation, there were no significant differences between groups in F0 maternal feed consumption values.
Food efficiency:
no effects observed
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):
effects observed, treatment-related
Description (incidence and severity):
Post-dose rooting is considered to be a behavioral response to taste aversion to the dosing formulations and not a toxic sign. Since there was a dose-related increase in the incidence of post-dose rooting (0, 2 and 3 females in the 0, 100 and 1000 mg/kg/day groups, respectively), it was presumed that the increasing concentrations across groups caused the adverse taste reaction.
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
At necropsy, mean final body weights of the F0 females were equivalent across all dose groups. The absolute weight and weight relative to final body weight of the paired ovaries were significantly reduced at 1000 mg/kg/day.
Gross pathological findings:
no effects observed
Description (incidence and severity):
There were no treatment-related macroscopic findings.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
There were no treatment-related microscopic findings.
Histopathological findings: neoplastic:
not examined
Other effects:
not examined
Number of abortions:
no effects observed
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
no effects observed
Early or late resorptions:
no effects observed
Dead fetuses:
no effects observed
Changes in pregnancy duration:
no effects observed
Changes in number of pregnant:
no effects observed
Other effects:
not examined
Details on maternal toxic effects:
During the post mating period, there were 9, 9, and 8 females in the 0, 100, and 1000 mg/kg/day dose groups, respectively, that were determined to be sperm positive; however, the total number of females confirmed pregnant at study completion was 9, 10 and 9, respectively. One pregnant female in the 100 mg/kg/day group was not identified as being sperm positive; one female each in the control and 1000 mg/kg/day groups was not pregnant, and one pregnant female at 1000 mg/kg/day did not deliver a litter. There was a statistically significant increase in precoital interval at 1000 /mg/kg/day although the increase was only approximately one day longer than the controls.
Key result
Dose descriptor:
NOAEL
Effect level:
100 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
other: maternal toxicity
Key result
Dose descriptor:
NOAEL
Effect level:
> 1 000 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
other: developmental toxicity
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects: no effects

Details on embryotoxic / teratogenic effects:
There was no evidence of F1 offspring toxicity at any dose.
Key result
Dose descriptor:
NOAEL
Effect level:
> 1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: There was no evidence of F1 offspring toxicity at any dose.
Abnormalities:
no effects observed
Developmental effects observed:
no

Summary of F1 Offspring Toxicity

F1

Bisphenol A Dianhydride (mg/kg/day)

0

100

1000

No. Live Litters

Postnatal Day 0

9

10

8

Postnatal Day 4

9

10

8

Average No. of Live Pups per Litter (pnd 0)

14.8 ± 1.2

12.7 ± 1.7

13.4 ± 1.1

Average No. of Live Pups per Litter (pnd 4)

14.6 ± 1.1

12.3 ± 1.7

13.3 ± 1.2

Average Pup Body Weight (g) per Litter (pnd 0)

6.33 ± 0.14

7.03 ± 0.17**

6.40 ± 0.14

Average Male Body Weight (g) per Litter (pnd 0)

6.45 ± 0.17

7.16 ± 0.19*

6.49 ± 0.16

Average Female Body Weight (g) per Litter (pnd 0)

6.22 ± 0.13

6.91 ± 0.17**

6.30 ± 0.13

Average Pup Body Weight (g) per Litter (pnd 4)

9.96 ± 0.33

11.22 ± 0.52

10.81 ± 0.51

Average Male Body Weight (g) per Litter (pnd 4)

10.20 ± 0.36

11.44 ± 0.58

11.02 ± 0.52

Average Female Body Weight (g) per Litter (pnd 4)

9.71 ± 0.30

10.93 ± 0.54

10.63 ± 0.52

% Percent Male Pups per Litter (pnd 0)

54.9 ± 5.1

48.5 ± 3.5

44.4 ± 3.9

% Percent Male Pups per Litter (pnd 4)

55.0 ± 5.2

60.2 ± 6.5

48.8 ± 2.7

* p = 0.05; ** p = 0.01

Conclusions:
Under the conditions of this study, the NOAEL for the F0 systemic toxicity was 100 mg/kg/day. The NOAEL for F0 reproductive toxicity was >1000 mg/kg/day for both sexes. The NOAEL for F1 offspring toxicity was > 1000 mg/kg/day.
Executive summary:

A screening reproduction/development test was conducted using methods comparable to OECD guideline 421 under GLP conditions (RTI International, 2005).

Sprague-Dawley male and female rats were exposed to the test material via gavage in corn oil at dose levels of 0, 100, and 1000 mg/kg/day (10/sex/group). Due to the limited toxicity observed in previous studies for a similar test substance, only two BPA-DA dose groups were used. Females were dosed for ~7 weeks (2 weeks prior to breeding, 2 week mating period, 3 week gestation period and lactation through postnatal day 4).

Minimal systemic toxicity was present in parental animals through the course of the study at 1000 mg/kg/day. In the F0 females, the only adverse effects were decreases in high dose body weight change (Days 7 to 14) and treatment related clinical signs at the high dose. At the high dose there was an increase in the pre-coital interval, but no effect on fertility. There was no evidence of reproductive toxicity in the F0 females at any dose, or any toxicity in the F1 offspring. At necropsy, for the F0 females there were no treatment effects with the exception of decreases in the absolute and relative paired ovary weights. 

Under the conditions of this study, the NOAEL for the F0 systemic toxicity was 100 mg/kg/day. The NOAEL for F0 reproductive toxicity was > 1000 mg/kg/day for both sexes. The NOAEL for F1 offspring toxicity was > 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:
other: The NOAEL was determined to be >1000 mg/kg in both rats and rabbits
Quality of whole database:
Three studies are available to address this endpoint. All were conducted under GLP conditions using methodology equivalent to standardised guidelines. The quality of the database is therefore considered to be high.
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

A screening reproduction/development test was conducted using methods comparable to OECD guideline 421 under GLP conditions (RTI International, 2005). The study was awarded a reliability score of 1 in accordance with the criteria set forth by Klimisch et al. (1997).

Sprague-Dawley male and female rats were exposed to the test material via gavage in corn oil at dose levels of 0, 100, and 1000 mg/kg/day (10/sex/group). Due to the limited toxicity observed in previous studies for a similar test substance, only two BPA-DA dose groups were used. Females were dosed for ~7 weeks (2 weeks prior to breeding, 2 week mating period, 3 week gestation period and lactation through postnatal day 4).

Minimal systemic toxicity was present in parental animals through the course of the study at 1000 mg/kg/day. In the F0 females, the only adverse effects were decreases in high dose body weight change (Days 7 to 14) and treatment related clinical signs at the high dose. At the high dose there was an increase in the pre-coital interval, but no effect on fertility. There was no evidence of reproductive toxicity in the F0 females at any dose, or any toxicity in the F1 offspring. At necropsy, for the F0 females there were no treatment effects with the exception of decreases in the absolute and relative paired ovary weights. Based on these results, the NOAEL for the F0 systemic toxicity was 100 mg/kg/day. The NOAEL for F0 reproductive toxicity was >1000 mg/kg/day for both sexes. The NOAEL for F1 offspring toxicity was >1000 mg/kg/day. 

The developmental toxicity potential of BPA-DA in the rabbit was investigated in a study conducted using methodology equivalent to OECD 414 and EPA OPPTS 870.3700 under GLP conditions (Hazleton Laboratories America, Inc., 1983). The study was awarded a reliability score of 1 in accordance with the criteria set forth by Klimisch et al. (1997).

Sixteen New Zealand White rabbits were dosed with the test material at 1000 mg/kg bw by gavage in CMC (carboxymethyl cellulose) on Days 6 through 18 of gestation. A further 16 were dosed with the vehicle alone.

A reduction in maternal bodyweight was seen at 1000 mg/kg. There were no effects on any foetal parameters from BPA-DA treatment and the NOAEL for developmental toxicity was considered to be ≥1000 mg/kg bw.

Based on the results of this study, BPA-DA is not a developmental toxin.

 

The developmental toxicity potential of BPA-DA in the rat was investigated in a study conducted using methodology equivalent to OECD 414 and EPA OPPTS 870.3700 under GLP conditions (Hazleton Laboratories America, Inc., 1987). The study was awarded a reliability score of 1 in accordance with the criteria set forth by Klimisch et al. (1997).

Twenty-four Sprague-Dawley rats were dosed with the test material at 1000 mg/kg bw by gavage in CMC (carboxymethyl cellulose) on Days 6 through 15 of gestation. A further 24 were dosed with the vehicle alone.

A reduction in maternal bodyweight was seen at 1000 mg/kg. There were no effects on any foetal parameters from BPA-DA treatment and the NOAEL for developmental toxicity was considered to be ≥1000 mg/kg bw.

Based on the results of this study, BPA-DA is not a developmental toxin.

Justification for selection of Effect on developmental toxicity: via oral route:

No key study was selected on the basis that several key studies have been provided to address this endpoint.  

Justification for classification or non-classification

In accordance with the criteria for classification as defined in Annex I, Regulation (EC) No 1272/2008, the substance does not require classification with respect to reproductive or developmental toxicity.