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Toxicological information

Toxicity to reproduction

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Administrative data

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

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2005
Report Date:
2005

Materials and methods

Test guideline
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

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Bisphenol A Dianhydride (BPA-DA; CAS RN 38103-06-9)

Test animals

Species:
rat
Strain:
other: CD® (Sprague-Dawley)
Sex:
male/female

Administration / exposure

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
Doses / concentrationsopen allclose all
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

Examinations

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.

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

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 / performance (P0)

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.

Effect levels (P0)

open allclose all
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.

Results: F1 generation

General toxicity (F1)

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

Developmental neurotoxicity (F1)

Behaviour (functional findings):
not examined

Developmental immunotoxicity (F1)

Developmental immunotoxicity:
not examined

Details on results (F1)

There was no evidence of F1 offspring toxicity at any dose.

Effect levels (F1)

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.

Overall reproductive toxicity

Reproductive effects observed:
no

Any other information on results incl. tables

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

Applicant's summary and conclusion

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.