4,4'-[(isopropylidene)bis(p-phenyleneoxy)]diphthalic dianhydride

Endpoint summary

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

Reference

Reference 1

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

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	8^a
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 ± 2.91^b
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. 

Reference 2

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

Reference

Reference 1

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

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

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 or test system 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.

Reference 2

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

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

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 or test system 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.

Reference 3

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

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.