[No public or meaningful name is available]

Toxicity to reproduction

Administrative data

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

23 Augustus - 09 November 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

Version / remarks:

2011

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: EPA, Health Effects Test Guidelines OPPTS 870.3650, Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test, July 2000.

Version / remarks:

2000

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of (1-methylethylidene)bis(4,1-phenyleneoxy-2,1-ethanediyl) bismethacrylate and 2-{4-[2-(4-{2-[2-(methacryloyloxy)ethoxy]ethoxy}phenyl)propan-2-yl]phenoxy}ethyl methacrylate

EC Number:

939-702-5

Molecular formula:

C23H24O4 (C2H4O)n

IUPAC Name:

Reaction mass of (1-methylethylidene)bis(4,1-phenyleneoxy-2,1-ethanediyl) bismethacrylate and 2-{4-[2-(4-{2-[2-(methacryloyloxy)ethoxy]ethoxy}phenyl)propan-2-yl]phenoxy}ethyl methacrylate

Test material form:

other: Clear colourless highly viscous liquid

Details on test material:

- Name of test material (as cited in study report): 2 moles ethoxylated bisphenol A dimethacrylate

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
Nulliparous and nonpregnant females and untreated animals were used at initiation of the study.
- Age at study initiation: Approximately 11 weeks.
- Weight at study initiation: mean weight at start of treatment was 306-309 gr (males) or 213 - 220 gr (females).
- Fasting period before study: no
- Housing:
Pre-mating: Animals were housed in groups of 5 animals/sex/cage in Macrolon cages.
Mating: Females were caged together with males on a one-to-one-basis in Macrolon cages.
Post-mating: Males were housed in their home cage with a maximum of 5 animals/cage. Females were individually housed in Macrolon cages. Pups were kept with the dam until termination
General: Sterilised sawdust as bedding material and paper as cage enrichment were supplied.
- Diet: Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).
- Water: Free access to tap water.
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
Environmental controls for the animal room were set to maintain 18 to 24°C, a relative humidity of 40 to 70%, approximately 15 room air changes/hour, and a 12-hour light/12-hour dark cycle. Any variations to these conditions were maintained in the raw data and had no effect on the outcome of the study.

IN-LIFE DATES
From: 23 Augustus to 09 November 2013

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Details on exposure:

- Method of formulation: Formulations (w/w) were prepared daily within 5 hours prior to dosing and were homogenized to a visually acceptable level. Adjustment was made for specific gravity of the test substance and the vehicle. No correction was made for the purity/composition of the test substance.
- Storage conditions of formulations: At ambient temperature.
- Justification for use and choice of vehicle (if other than water): Based on trial formulations performed at WIL Research Europe.
- Dose volume: 5 mL/kg body weight. Actual dose volumes were calculated according to the latest body weight.

Details on mating procedure:

- M/F ratio per cage:1/1 (one female was cohabitated with one male of the same treatment group, avoiding sibling mating.
- Length of cohabitation: A maximum of 14 days was allowed for mating.
- Proof of pregnancy: Detection of mating was confirmed by evidence of sperm in the vaginal lavage, by staging of the estrous cycle and/or or by the appearance of an intravaginal copulatory plug. This day was designated Day 0 post-coitum. Once mating had occurred, the males and females were separated.
-Further matings after two unsuccessful attempts: no
- After successful mating each pregnant female was caged individually in Macrolon cages (MIII type, height 18 cm).

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The delegated phase was performed by the Principal Investigator for Formulation Analysis.

Samples (0.5 mL) were taken using a pipette (a clean pipette tip was used for every group), and were weighed on an analytical balance at 4 decimals precision. During sampling, formulations were placed on a magnetic stirrer. Immediately after sampling (accuracy and homogeneity samples) or after 5 hours at room temperature under normal laboratory light conditions (stability samples), samples were stored on dry ice. Samples remained on dry ice until receipt at ABL, The Netherlands.

Samples of formulations were analyzed for homogeneity (highest and lowest concentration) and accuracy of preparation (all concentrations). Stability in vehicle over 5 hours at room temperature under normal laboratory light conditions was also determined (highest and lowest concentration).

The accuracy of preparation was considered acceptable if the mean measured concentrations were 90-110% of the target concentration. Homogeneity was demonstrated if the coefficient of variation was = 10%. Formulations were considered stable if the relative difference before and after storage was maximally 10%.

Formulation samples were directly shipped instead of being sent to the ‘weegkamer’ first.
Evaluation: The test site received the samples in good condition; the study was not affected.

Duration of treatment / exposure:

Males were exposed for 29 days, i.e. 2 weeks prior to mating, during mating, and up to the day prior to the scheduled necropsy. Females were exposed for 43-46 days, i.e. during 2 weeks prior to mating, during mating, during post-coitum, and during at least 4 days of lactation (up to the day prior to scheduled necropsy).Three females( from Group 1, 2 and 3 respectively) and 2 females from Group 4 were not dosed during littering.

Pups were not dosed directly but could have potentially be exposed to the test substance in utero, via maternal milk or from exposure to maternal urine/faeces.

Frequency of treatment:

Once daily for 7 d/w.

Details on study schedule:

- Age at mating of the animals in the study: Approximately 13 weeks

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

62 mg/kg bw/day

Doses / concentrations 2

Dose / conc.:

250 mg/kg bw/day

Doses / concentrations 3

Dose / conc.:

1 000 mg/kg bw/day

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Dose levels were based on results of a 14-Day dose range finding study (See attached results).
- Based on the results of this range finding study, dose levels for the main study were: 62, 250 and 1000 mg/kg.

- Since no clinical signs were observed in the range finding study, clinical observations in the main study were started at no specific time point but within a similar time period for all animals after dosing.

- Selection of animals for selected measurements: 5 animals/sex/group were randomly selected at allocation for functional observations, clinical pathology, macroscopic examination (full list), organ weights (full list) and histopathology (see also respective paragraphs). Only females with live offspring were selected.
- Parturition: The females were allowed to litter normally. Day 1 of lactation was defined as the day when a litter was found completed (i.e. membranes, placentas cleaned up, nest built up and/or feeding of pups started). Females that were littering were left undisturbed.
- Identification of pups: On Day 1 of lactation, all pups were individually identified by means of subcutaneous injection of Indian ink.

Positive control:

Not required.

Examinations

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS
- Time schedule: At least twice daily.

DETAILED CLINICAL OBSERVATIONS
- Time schedule: Daily from treatment onwards up to the day prior to necropsy, detailed clinical observations were conducted for all animals at no specific time point, but within a similar time period after dosing for the respective animals. Once prior to start of treatment and at weekly intervals during the treatment period this was also performed outside the home cage in a standard arena.
The time of onset, grade and duration of any observed sign was recorded.

BODY WEIGHT
- Time schedule for examinations: Males and females were weighed on the first day of exposure and weekly thereafter. Mated females were weighed on Days 0, 4, 7, 11, 14, 17 and 20 post-coitum, and during lactation on Days 1 and 4.

FOOD CONSUMPTION
- Weekly, for males and females. Food consumption was not recorded during the mating period. Food consumption of mated females was measured on Days 0, 4, 7, 11, 14, 17 and 20 post-coitum and on Days 1 and 4 of lactation.

FOOD EFFICIENCY: yes

WATER CONSUMPTION
No. Subjective appraisal was maintained during the study, but no quantitative investigation introduced as no effect was suspected.

OPHTHALMOSCOPIC EXAMINATION
No

HAEMATOLOGY
- Time schedule for collection of blood: prior to scheduled post mortem examination (between 7.00 and 10.30 a.m.).
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes (with a maximum of 24 hours). Water was provided.
- How many animals: 5 animals/sex/group
- Parameters checked were: According to test guidelines

CLINICAL CHEMISTRY
- Time schedule for collection of blood: prior to scheduled post mortem examination (between 7.00 and 10.30 a.m.).
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes (with a maximum of 24 hours). Water was provided.
- How many animals: 5 animals/sex/group
- Parameters checked were: According to test guidelines

THYROID HORMONE ANALYSIS
- Time schedule for collection of blood: prior to scheduled post mortem examination (between 7.00 and 10.30 a.m.).
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes (with a maximum of 24 hours). Water was provided.
- How many animals: 5 animals/sex/group
- Parameters checked were: According to test guidelines
- Instead of taking 1 mL of blood for the serum sample from males, 0.5 mL was initially taken and an additional 0.5 mL had to be collected again. Animals. 3 and 5 (Group 1) had already been necropsied, so an additional 0.5 mL could not be collected.
Evaluation: The extra collection has no effect on the study and sufficient data is available for the insufficient samples from animals 3 and 5.

URINALYSIS
No

NEUROBEHAVIOURAL EXAMINATION
- Time schedule for examinations: The selected males were tested during Week 4 of treatment and the selected females were tested during lactation (all before blood sampling).
- Dose groups that were examined: all
- Battery of functions tested: According to test guidelines
- Motor activity and functional observation assessment were performed on Day 3 of lactation for one Group 4 animal instead of from Day 4.
Evaluation: Conducting these tests one day too early has no impact on the results.

GENERAL REPRODUCTION DATA
- Male number paired with, mating date, confirmation of pregnancy, and delivery day was recorded.
- Pregnant females were examined to detect signs of difficult or prolonged parturition, and cage debris of these females was examined to detect signs of abortion or premature birth.
- Any deficiencies in maternal care (such as inadequate construction or cleaning of the nest, pups left scattered and cold, physical abuse of pups or apparently inadequate lactation or feeding) were examined.

Oestrous cyclicity (parental animals):

Not determined.

Sperm parameters (parental animals):

Slides of the testes were prepared for histopathological staging of spermatogenesis (5 males of the control and high dose group and animals suspected to be infertile).

Litter observations:

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross abnomalies, weight gain, physical or behavioural abnormalities.

- Mortality: The numbers of live and dead pups on Day 1 of lactation and daily thereafter were determined. If possible, defects or cause of death were evaluated.
- Clinical signs: At least once daily, detailed clinical observations were made in all animals.
During the lactation period, no clinical observations were entered online for pups 13 -15 of 1 Group 2 litter on Day 6 and for pups 13 +14 of an other Group 2 on Day 5.
Evaluation: Sufficient data is available for a thorough evaluation.

- Body weights: Live pups were weighed on Days 1 and 4 of lactation.
- Sex: Sex was determined for all pups on Days 1 and 4 of lactation (by assessment of the ano-genital distance).

GROSS EXAMINATION OF DEAD PUPS
Yes, if possible, defects or cause of death were evaluated.

Postmortem examinations (parental animals):

GROSS PATHOLOGY
All animals were fasted overnight (with a maximum of 24 hours) prior to planned necropsy, but water was provided. Animals surviving to scheduled necropsy and all moribund animals were deeply anaesthetised and subsequently exsanguinated.
- The number of former implantation sites and corpora lutea were recorded for all paired females.
- Selected 5 animals/sex/group and all animals that died spontaneously or were killed in extremis: According to test guidelines
- All remaining animals and females which failed to deliver: According to test guidelines

ORGAN WEIGHTS
- Selected 5 animals/sex/group: According to test guidelines
- All remaining males: Epididymides and testes

- Inadvertently, the testes and epididymides from 1 Group 2 animal were not weighed at necropsy, and the rectum from 1 Group 1 animal and the mesenteric lymph node from 1 Group 4 animal were not available for histopathology because the tissues were not discernible at necropsy or trimming, or were erroneously not collected.
Evaluation: Sufficient data was available for evaluation.

HISTOPATHOLOGY
- According to test guidelines
- The skin was collected from all Group 1 and 2 animals at necropsy.
Evaluation: The tissue was stored but not analyzed further. This has no influence on the study.

Postmortem examinations (offspring):

SACRIFICE
Pups surviving to planned termination were killed by decapitation on lactation Days 5-7.

GROSS NECROPSY
All pups were sexed and descriptions of all external abnormalities were recorded. The stomach was examined for the presence of milk. If possible, defects or cause of death were evaluated. Any abnormal pup, organ or tissue was preserved in 10% buffered formalin for possible further examination.

HISTOPATHOLOGY / ORGAN WEIGHTS
No.

Statistics:

The following statistical methods were used to analyze the data:
- If the variables could be assumed to follow a normal distribution, the Dunnett-test (many-to-one t-test) based on a pooled variance estimate was applied for the comparison of the treated groups and the control groups for each sex.
- The Steel-test (many-to-one rank test) was applied if the data could not be assumed to follow a normal distribution.
- The Fisher Exact-test was applied to frequency data.
- The Kruskal-Wallis nonparametric ANOVA test was applied to motor activity data to determine intergroup differences.

All tests were two-sided and in all cases p < 0.05 was accepted as the lowest level of significance.
Group means were calculated for continuous data and medians were calculated for discrete data (scores) in the summary tables. Test statistics were calculated on the basis of exact values for means and pooled variances. Individual values, means and standard deviations may have been rounded off
before printing. Therefore, two groups may display the same printed means for a given parameter, yet display different test statistics values.

Reproductive indices:

For each group, the following calculations were performed:
- Mating index: Number of females mated/Number of females paired x 100
- Fertility index: Number of pregnant females/Number of females paired x 100
- Conception index: Number of pregnant females/Number of females mated x 100
- Gestation index: Number of females bearing live pups/Number of pregnant females x 100
- Duration of gestation: Number of days between confirmation of mating and the beginning of parturition

Offspring viability indices:

- Percentage live males at First Litter Check: Number of live male pups at First Litter Check/Number of live pups at First Litter Check x 100
- Percentage live females at First Litter Check: Number of live female pups at First Litter Check/Number of live pups at First Litter Check x 100
- Percentage of postnatal loss Days 0-4 of lactation: Number of dead pups on Day 4 of lactation/Number of live pups at First Litter Check x 100
- Viability index: Number of live pups on Day 4 of lactation / Number of pups born alive x 100
- Pre-implantation loss (%): (number of corpora lutea - number of implantation sites)/number of corpora lutea x 100
- Post-implantation loss (%): (number of implantation sites - number of live fetuses) / number of implantation sites x 100

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

No adverse clinical signs of toxicity were noted during the observation period.
Salivation was seen for all animals at 1000 mg/kg, for four males and one female at 250 mg/kg and for eight males at 62 mg/kg. This was considered to be a physiological response rather than a sign of systemic toxicity considering the nature and minor severity of the effect and its time of occurrence (i.e. after dosing). This sign may be related taste or possible irritancy of the test substance, and is not reflective of systemic toxicity.
Incidental findings noted included chromodacryorrhea and opacity of the right eye, noted for a single control male. These findings occurred within the range of background findings to be expected for rats of this age and strain which are housed and treated under the conditions in this study. As these were noted for a control male, they were not treatment related.

Mortality:

mortality observed, non-treatment-related

Description (incidence):

One Group 4 female no. 79 was found dead the morning after pairing began. She was partially cannibalized with her vagina, urinary bladder, rectum and clitoral glands missing. Severe necrosis of the trachea was found at the microscopic examination, indicating her death was secondary to a gavage error and was not attributable to treatment.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

No toxicologically relevant changes in body weights and body weight gain were noted up to 1000 mg/kg.
Body weight gain was slightly lower for males at 250 and 1000 mg/kg on Day 8 of the premating period. The difference from controls was only very slight and moreover, no statistically significant decrease in body weight for males was observed during the premating period for males. Taken together, it was not considered to be toxicologically relevant.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

Food consumption before or after allowance for body weight was similar between treated and control animals.

Ophthalmological findings:

not examined

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

There were no toxicologically relevant effects on haematology parameters up to 1000 mg/kg.
The significant increase in mean corpuscular volume (MCV) and mean corpuscular haemoglobin (MCH) seen for males at 62 and 250 mg/kg. These parameters were unaffected for females.
The significant decrease in eosinophils was seen for females at 250 mg/kg only. A trend towards an increase in eosinophils with increasing doses was seen for males, but changes were not statistically significant. All these changes occurred in the absence of a dose response effect and remained within the range considered normal for animals of this age and strain. As such, these differences from controls were not considered to be toxicologically relevant.

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

Cholesterol was significantly higher than controls for males at 250 and 1000 mg/kg and for females in all treated groups. Both sexes show a clear dose response effect. However, in the absence of toxicologically significant findings in any other parameters, it was not considered to be toxicologically relevant.
The significant decrease in albumin was noted for males (all treated groups); the same decrease dose-related decrease was seen for females, but was not statistically significant.
The significant decrease in creatinine seen for females (62 and 250 mg/kg) did not show a dose-dependent trend as females at 1000 mg/kg were not affected. There were no creatinine changes for males. Therefore, this change was not considered to be toxicologically relevant as it occurred in the absence of a treatment related distribution and remained within the range considered normal for rats of this age and strain.
Males at 62 mg/kg had a significant increase in the thyroid hormone total thyroxine (T4) compared to controls. This was also not considered to be toxicologically relevant as it occurred in the absence of a treatment related distribution and no corroborative effects were seen microscopically.

Endocrine findings:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

no effects observed

Description (incidence and severity):

Hearing ability, pupillary reflex, static righting reflex and grip strength were normal in all selected animals.
The variation in motor activity did not indicate a relation with treatment.
All groups showed a similar habituation profile with very high activity in the first interval that decreased over the duration of the test period.

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Treatment-related microscopic findings were found in female thyroid glands and comprised:
Diffuse follicular hypertrophy/hyperplasia that was seen in 2/5 males (minimal) and 1/5 females (slight) of Group 2, 2/5 males (minimal) of Group 3 and 1/5 males (slight) and 3/5 females (minimal) of Group 4. However, this remained within the normal background range of findings for rats of this age and strain, and based on the absence of a dose-relationship, the diffuse follicular hypertrophy/hyperplasia was considered to be adaptive in nature and not toxicologically relevant.
The assessment of the integrity of the spermatogenetic cycle did not provide any evidence of impaired spermatogenesis.
There were no morphological findings in the reproductive organs of either sex which could be attributed to the test item.
All microscopic findings recorded were considered to be within the normal range of background pathology encountered in rats of this age and strain and in this type of study.

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):

No toxicologically relevant effects on reproductive parameters were noted.
The mating, fertility and conception indices, precoital time, and number of corpora lutea and implantation sites were unaffected by treatment. There were no toxicologically relevant effects on the gestation index and duration, parturition or maternal care. There were 9, 10, 10 and 9 pregnant females in the control, 62, 250 and 1000 mg/kg groups, respectively.
When recalculated without counting the single implantation site for one control female, the mean number of implantation sites for controls was 13.3. Even discounting this female, there were no differences between control and treated females.
For one Group 3 female, the number of pups (14) was slightly higher than the number of implantations (13). This was considered to be caused by normal resorption of implantation sites as these enumerations were performed on Day 7 of lactation
The gestation index and duration of gestation were unaffected by treatment up to 1000 mg/kg.
No signs of difficult or prolonged parturition were noted among the pregnant females.
Examination of cage debris of pregnant females revealed no signs of abortion or premature birth and no deficiencies in maternal care were observed. There were 8, 10, 10 and 9 females with living pups on Day 1 of lactation. One Group 1 female (control) was found with one giant fetus in the uterus at the macroscopic examination, where the fetus was so large it would not be possible for the mother to deliver. One Group 4 Female (1000 mg/kg) was found dead the day after the mating period began.

Effect levels (P0)

Key result

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Remarks on result:

not determinable due to absence of adverse toxic effects

Target system / organ toxicity (P0)

Critical effects observed:

no

Results: F1 generation

General toxicity (F1)

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

Incidental clinical signs of pups included pale appearance and blue spot of the abdomen, which were noted for a single pup at 250 mg/kg. The nature and incidence of these signs remained within the range considered normal for pups of this age, and were not considered toxicologically relevant. No clinical signs were noted for any other pup.

Mortality / viability:

mortality observed, non-treatment-related

Description (incidence and severity):

Two pups (from separate litters) at 1000 mg/kg were missing on Day 2. They were most likely cannibalized. There were no differences in Day 1 body weights between these pups and control pups. No toxicological relevance was attributed to these dead/missing pups since the incidence remained within the range considered normal for pups of this age. There were no other pups that died or went missing in the control or other treated groups.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

Body weights of pups were unaffected by treatment up to 1000 mg/kg.

Food consumption and compound intake (if feeding study):

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

Anogenital distance (AGD):

not examined

Nipple retention in male pups:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

effects observed, non-treatment-related

Description (incidence and severity):

The only macroscopic finding noted was blue spot on the abdomen, noted for a single pup at 250 mg/kg. Due to the nature of this finding and its limited incidence, it was not considered to be toxicologically relevant.

Histopathological findings:

not examined

Developmental neurotoxicity (F1)

Behaviour (functional findings):

not examined

Developmental immunotoxicity (F1)

Developmental immunotoxicity:

not examined

Details on results (F1)

Number of dead and living pups at first litter check, postnatal loss, viability index and sex ratio were unaffected by treatment, and clinical signs, body weight and external macroscopy did not reveal treatment-related findings.

Effect levels (F1)

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Remarks on result:

not determinable due to absence of adverse toxic effects

Target system / organ toxicity (F1)

Critical effects observed:

no

Overall reproductive toxicity

Reproductive effects observed:

no

Any other information on results incl. tables

No test substance was detected in the Group 1 formulations from the week 1, 3 and 6 preparations.

 

The concentrations analyzed in the formulations of Groups 2, 3 and 4 in weeks 1, 3 and 6 were in

agreement with target concentrations (i.e. mean accuracies between 90% and 110%).

 

The formulations of Group 2 and 4 were homogeneous (i.e. coefficient of variation = 10%).

Formulations over the entire range were stable when stored at room temperature under normal

laboratory light conditions for at least 5 hours (i.e. relative difference = 10%).

 

The long term storage samples were stable at =-70°C for at least 21 days.

Applicant's summary and conclusion

Conclusions:

In conclusion, treatment with 2 moles ethoxylated bisphenol A dimethacrylate by oral gavage in male and female Wistar Han rats at dose levels of 62, 250 and 1000 mg/kg revealed no parental, developmental or reproductive toxicity up to 1000 mg/kg.

Based on these results, a parental, reproduction and developmental No Observed Adverse Effect Level (NOAEL) of at least 1000 mg/kg was derived.

Executive summary:

Combined 28-day repeated dose toxicity study with the reproduction/developmental toxicity screening test was performed in rats with 2 moles ethoxylated bisphenol A dimethacrylate by oral gavage (OECD guideline 422).

Based on the results of a 14-day dose range finding study, the dose levels for this study were 62, 250 and 1000 mg/kg. After acclimatization, four groups of ten male and ten female Wistar Han rats were exposed by oral gavage to the test substance. Males were exposed for 29 days, i.e. 2 weeks prior to mating, during mating, and up to termination. Females were exposed for 43-46 days, i.e. during 2 weeks prior to mating, during mating, during postcoitum, and during at least 4 days of lactation.

The following observations and examinations were evaluated: mortality / viability, clinical signs (daily),

functional observations and locomotor activity (end of treatment), body weight and food consumption (at least at weekly intervals), clinical pathology (end of treatment), macroscopy at termination, organ weights and histopathology on a selection of tissues, and reproduction/developmental parameters, consisting of mating, fertility and conception indices, precoital time, number of corpora lutea and implantation sites, gestation index and duration, parturition, maternal care, sex ratio and early postnatal pup development (mortality, clinical signs, body weights and macroscopy). Formulations were analyzed once during the study to assess accuracy, homogeneity and stability.

 

Accuracy, homogeneity and stability of formulations were demonstrated by analyses.

No parental toxicity was observed up to the highest dose level tested (1000 mg/kg). No toxicologically relevant changes were noted in any of the parental parameters investigated in this study (i.e. clinical appearance, functional observations, body weight, food consumption, clinical laboratory investigations, macroscopic examination, organ weights, and microscopic examination).

No reproductive toxicity was observed up to the highest dose level tested (1000 mg/kg).

No developmental toxicity was observed up to the highest dose level tested (1000 mg/kg).

No treatment-related changes were noted in any reproductive or developmental parameters investigated in this study (i.e. mating, fertility and conception indices, precoital time, and numbers ofcorpora lutea and implantation sites, gestation index and duration, parturition, maternal care and early postnatal pup development consisting of mortality, clinical signs, body weight and macroscopy).

Treatment with 2 moles ethoxylated bisphenol A dimethacrylate by oral gavage in male and female Wistar Han rats at dose levels of 62, 250 and 1000 mg/kg revealed no parental, developmental or reproductive toxicity up to 1000 mg/kg. Based on these results, a parental, reproduction and developmental No Observed Adverse Effect Level (NOAEL) of at least 1000 mg/kg was derived.