reaction products of diglycidyl ether bisphenol F (DGEBF) and oligomeric phenol diglycidyl ethers with acrylic acid

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study, GLP

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test) (adotped 22nd March 1996)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: OPPTS 870.3650, July 2000

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

reaction products of diglycidyl ether bisphenol F (DGEBF) and oligomeric phenol diglycidyl ethers with acrylic acid

EC Number:

700-487-6

Molecular formula:

Not applicable.

IUPAC Name:

reaction products of diglycidyl ether bisphenol F (DGEBF) and oligomeric phenol diglycidyl ethers with acrylic acid

Details on test material:

- Name of test material (as cited in study report): Epoxy half acrylate
- Physical state: liquid
- Analytical purity: 100%, UVCB

Test animals

Species:

rat

Strain:

other: RccHan: WIST(SPF)

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 12 weeks (Males) / 11 weeks (Females)
- Weight at study initiation: Males: 323 - 396 g / Females: 160 - 237 g
- Housing: Individually in Makrolon type-3 cages or type-4 (for animals over 400 g) with wire mesh tops and sterilized standard softwood bedding with paper enrichment / During the pre-pairing period, cages with males were interspersed amongst those holding females to promote the development of regular estrus cycles. During pairing females were housed with males (1:1) in Makrolon type-3 cages.
- Diet (e.g. ad libitum): Pelleted standard Harlan Teklad 2018C rodent maintenance diet, ad libitum
- Water (e.g. ad libitum): community tap-water, ad libitum
- Acclimation period: 9 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30 - 70
- Air changes (per hr): 10 - 15
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: PEG300

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
- dose formulations were prepared weekly using the test item as supplied
- the test item was weighed into a glass beaker and the required amount of vehicle was added (w/v). Using a magnetic stirrer, a homogeneous suspension
was prepared. Separate formulations were prepared for each concentration. Homogeneity of the test item in the vehicle was maintained during the daily administration period using a magnetic stirrer.
- Dose formulations were stored in refrigerator (5 ± 3 °C) in glass beakers as daily aliquots

VEHICLE
- Justification for use and choice of vehicle (if other than water): not stated in report
- Concentration in vehicle: Group 1: 0 mg/mL / Group 2: 25 mg/mL / Group 3: 75 mg/mL / Group 3: 250 mg/mL
- Amount of vehicle (if gavage): 4 mL/kg body weight

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

- samples were delivered to the analytical department at ambient temperature and analyzed immediately or stored frozen (at -20 ± 5 °C) until analysis
- analysis by HPLC coupled to a UV detector

Details on mating procedure:

- Impregnation procedure: cohoused
- If cohoused:
- M/F ratio per cage: 1:1
- Length of cohabitation: up to 14 d
- After 14 days of unsuccessful pairing replacement of first male by another male with proven fertility.
- Further matings after two unsuccessful attempts: no
- Proof of pregnancy: vaginal plug / sperm in vaginal smear referred to as day 0 of pregnancy

Animals in recovery groups were not mated.

Duration of treatment / exposure:

Males: 42 days (during 14 days pre-pairing period, up to 28 days pairing period and after pairing period).
Females: Minimum 5 weeks (14 days pre-pairing period, up to 28 days pairing period, approximately 21 days of gestation period and lactation period to day 3 post partum).
Recovery Groups): 42 d

Frequency of treatment:

daily

Duration of test:

Males: 42 days / Females: Minimum 5 weeks

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 100, 300, 1000 mg/kg bw/d
Basis:
actual ingested

No. of animals per sex per dose:

12 (main groups) / 5 (recovery groups)

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: based on a previous dose range-finding study
- Rationale for animal assignment (if not random): Performed after at least three days of acclimatization using a computer-generated random algorithm. Body
weights (recorded on the day of allocation) were taken into consideration in order to ensure similar mean body weights in all groups.
- Rationale for selecting satellite groups: observation of reversibility, persistence or delayed occurrence of systemic toxic effects
- Post-exposure recovery period in satellite groups: 20 d (recovery groups)

Examinations

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Viability / Mortality: Twice daily; cage-side clinical observations (once daily, during acclimatization and up to day of necropsy; additionally females were observed for signs of difficult or prolonged parturition, and behavioral abnormalities in nesting and nursing
- Cage side observations: changes in skin, fur, eyes, mucous membranes, occurrence of secretions and excretions, and autonomic activity (e.g.lacrimation, piloerection, pupil size, unusual respiratory pattern). Changes in gait, posture and response to handling as well as the presence of clonic or tonic movements, stereotypes or bizarre behavior

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Males: once prior to the first administration of the test item and weekly thereafter.
Females: once prior to the first administration of the test item, weekly during the pre-pairing
and pairing and on days 0, 6, 13 and 20 post coitum

BODY WEIGHT: Yes
- Time schedule for examinations: Once during acclimatization and daily from treatment start to day of necropsy

FOOD CONSUMPTION):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
Males: Pre-pairing days 1 - 4, 4 - 8, 8 - 11 and 11 - 13 and weekly during after pairing.
Females: Pre-pairing days 1 - 4, 4 - 8, 8 - 11 and 11 - 13 and for periods: days 0 - 7, 7 – 14 and 14 - 21 post coitum and 1 - 4 post partum.
No food consumption was recorded during the pairing period.

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No

HAEMATOLOGY: Yes
- described in detail in IUCLID section "Repeated dose toxicity"

CLINICAL CHEMISTRY: Yes
- described in detail in IUCLID section "Repeated dose toxicity"

NEUROBEHAVIOURAL EXAMINATION: Yes
- described in detail in IUCLID section "Repeated dose toxicity"

POST-MORTEM EXAMINATIONS: Yes
Main Groups:
Males were sacrificed on the day after completion of the treatment when they were no longer necessary for the assessment of reproductive performance.
Dams and pups were sacrificed on day 4 post partum. If birth did not occur on the expected date (day 21 post coitum), the female was sacrificed and
examined on day 25 post coitum.
Recovery Groups:
Males and females were sacrificed after the 20-day recovery period.

GROSS PATHOLOGY: Yes (see table 1)
HISTOPATHOLOGY: Yes (see table 1)

Ovaries and uterine content:

- Gravid uterus weight: Not applicable (screening study)
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: No (screening study)
- Number of late resorptions: No (screening study)
- Other: duration of gestation, post-implantation loss, litter size at first litter check, postnatal loss days 0 - 4 post partum

Fetal examinations:

F1 Pups:
First Litter Check: Offspring were examined as soon as possible after completion of delivery for litter size, number of live and still births, and any gross abnormalities.
Viability / Mortality: Daily
Clinical Signs: Daily observations for any abnormal findings.
Sex: On days 0 (if possible), 1 and 4 post partum.
Body Weights: On days 0 (if possible), 1 and 4 post partum.

Statistics:

- Means and standard deviations of various data were calculated and included in the report.
- The Dunnett-test (many to one t-test) based on a pooled variance estimate was applied if the variables could be assumed to follow a normal distribution for the comparison of the treated groups and the control groups for each sex.
- The Steel-test (many-one rank test) was applied instead of the Dunnett-test when the data could not be assumed to follow a normal distribution.
- Fisher's exact-test was applied if the variables could be dichotomized without loss of information.

Indices:

percentage mating (number of females mated as a percentage of females paired)
fertility index (number of females achieving pregnancy as a percentage of females paired)
conception rate (number of females achieving pregnancy as a percentage of females mated)
gestation index (number of females with living pups as a percentage of females pregnant)
Birth index (number of pups born alive as a percentage of implantations)
Viability index (number of pups alive on day 4 post partum as a percentage of pups born alive)

Results and discussion

Results: maternal animals

Maternal developmental toxicity

Details on maternal toxic effects:

Maternal toxic effects:yes

Details on maternal toxic effects:
No test item-related deaths, clinical signs, findings during functional observational battery and locomotor activity measurement were noted during the study at any dose level.
The slight and reversible reduction in food consumption and body weight/body weight gain in the 1000 mg/kg bw/d dose group was considered not to be adverse.
No test item-related adverse effects on hematology or clinical biochemistry parameters were noted.
No test item-related changes in organ weights were noted females at any dose level.
Histopathological examination revealed test item-related lesions in the stomach (epithelial hyperplasia/hyperkeratosis of the non-glandular mucosa in forestomach) in females in the 1000 mg/kg bw/d dose group. This finding was correlated to crateriform retractions observed during the necropsy. Due to full reversibility following the treatment free period, these findings were considered not to be adverse.

Effect levels (maternal animals)

1

Dose descriptor:

NOEL

Effect level:

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

Based on:

act. ingr.

Basis for effect level:

other: other:

2

Dose descriptor:

NOAEL

Effect level:

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

Based on:

act. ingr.

Basis for effect level:

other: maternal toxicity

Results (fetuses)

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Fetal abnormalities

Abnormalities:

not specified

Overall developmental toxicity

Developmental effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

No test item-related deaths or clinical signs were noted during the study at any dose level. No test item-related findings were recorded during functional observational battery and locomotor activity measurement in males or females in any group.
Treatment with the test item caused a slight and reversible reduction in food consumption in males and females in group 4. This effect was considered not to be adverse.
In males in group 4, treatment with the test item caused a slight reduction in body weights and body weight gain observed during the most of the study. In females in group 4, a minor reduction in body weight gain was noted during the gestation period whereas absolute body weights were not changed. This effect was considered not to be adverse.
Treatment with the test item caused an increase in potassium concentration in males and females of group 4. This effect was considered not to be adverse. No further test item-related changes in hematology or clinical biochemistry parameters were noted in males or females at any dose level.
Treatment with the test item caused an increase in liver weights in males in groups 2, 3 and 4. No further test item-related changes in organ weights were noted in males or females at any dose level.
Treatment with the test item caused crateriform retractions in males in groups 3 and 4.
Histopathological examination revealed test item-related lesions in the stomach (epithelial hyperplasia/hyperkeratosis of the non-glandular mucosa in forestomach) in males in groups 3 and 4 and in females in group 4. This finding was correlated to crateriform retractions observed during the necropsy.
Further, vacuolation of epithelium was found in epididymides of males in groups 3 and 4 with dose-related incidence and severity. Minimal edema and decreased tubular size were found in cauda epididymis of males in group 4, occasionally apoptotic epithelial cells were present and the stroma around vacuolated tubules was slightly oedematous with infiltration of a few inflammatory cells. Edema with apparent decreased tubular size was observed within the cauda epididymis
Test item-related changes in the tubulo-alveoli of the ventral lobe in the prostate and minimal to slight vacuolation of the glandular epithelium and decreased secretory content were recorded in males of groups 2, 3 and 4 with the incidence and severity being unrelated to the dose levels. In the liver, centrilobular hepatocellular hypertrophy was recorded in males of groups 3 and 4 with a dose-related incidence and severity. The enlarged hepatocytes displayed a “ground glass” appearance.
Because of the type of these findings and their partial or full reversibility following the treatment free period, these findings were considered not to be adverse.

Reproduction and Breeding Data
Mating performance, fertility, duration of gestation, corpora lutea count, implantation rate, post implantation and post natal losses as well as litter size were not affected by the treatment with the test item at any dose level.

Litter Data - F1 Pups
No test item-related abnormal findings were noted in pups at first litter check or during lactation in any group. Sex ratios at first litter check were unaffected by exposure to the test item.
Pup body weights and body weight gain were not affected by the treatment with the test item in any group.
No test item-related macroscopical findings were noted during necropsy of pups in any group.

Based on these results the NOAEL for general toxicity was established at 1000 mg/kg bw/d.
The NOAEL for reproduction/developmental toxicity was considered to be 1000 mg/kg bw/d.

Executive summary:

In a Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test according to OECD guideline 422, adotped 22nd March 1996, and OPPTS 870.3650, July 2000, Epoxy half acrylate (100% a.i.) was administered to 12 RccHan: WIST(SPF) rats/sex/dose orally via gavage at dose levels of 0, 100, 300 and 1000 mg/kg bw/day in the main groups. A recovery group of 5 RccHan: WIST(SPF) rats/sex/dose in the highest dose group and the control group, which was not mated, was observed for reversibility, persistence or delayed occurrence of systemic toxic effects.

In the P generation, the test item was administered to male rats for 42 days and to female rats for 14 days prior to pairing, throughout the pairing and gestation periods until the F1 generation reached day 4 post partum. In recovery animals, the test item was administered to male and female rats for 42 days followed by a 20-day recovery period without treatment.

General Toxicity

No test item-related deaths, clinical signs or behavioral changes were noted in males or females during the study at any dose level.

A minor and reversible reduction in food consumption, body weights and body weight gain was observed in males at the high-dose level during the treatment and a recovery of these parameters after the completion of the treatment. Reduction in food consumption and body weight gain was also noted in females at the high-dose level but the effects were less pronounced in this gender.

Food consumption and body weight gain were reduced in females in main groups during the gestation period but without any effects on the absolute body weights. In the recovery groups only slight reduction in body weight gain was recorded, followed by a recovery after completion of the treatment. Food consumption and absolute body weights in the recovery animals were not affected by the treatment. Effects on food consumption and body weights were considered not to be adverse.

Treatment with the test item at the high-dose level caused an increase in potassium concentration in males and females in main groups after 14 days of treatment. Further, albumin concentration was increased whereas globulin concentration was decreased resulting in an increased albumin to globulin ratio in males and females in the recovery groups after the treatment for 42 days. These findings were reversible; no significant changes in the clinical biochemistry parameters were recorded after the 20-day treatment free period and therefore they were considered not to be adverse.

At terminal examination, crateriform retractions were found in males in main groups at the high and mid-dose levels. This finding was correlated to the epithelial hyperplasia and hyperkeratosis found on the non-glandular mucosa (forestomach). Changes in the stoma were considered to be indicative of direct contact irritancy. Further, erosion and ulcer(s) were observed in the glandular or non-glandular mucosa, which was considered to be secondary to the non-glandular mucosal findings. Because no macroscopical or microscopical findings were found in the stomach of animals terminated after the recovery period, these changes were reversible and therefore considered not to be adverse.

An increase in the liver weights in males was noted down to the low-dose level. At the high-dose level, this finding was correlated histologically with centrilobular hepatocellular hypertrophy.

These findings are suggestive of an adaptative response to mixed function oxidase induction.

Changes in the liver were reversible after completion of the treatment; they were not recorded for animals terminated after 20-day of the recovery period. Increased liver weights and hepatocellular hypertrophy were considered not to be adverse.

A reduced thymus weight was noted in females at the high-dose level after the recovery period. This finding was considered to possibly be due to the treatment with the test item. In the absence of any histopathological change in this organ, reduced thymus weight was considered not to be adverse.

 

Histopathological examination revealed also test item-related lesions in the prostate andepididymides in males. The pathogenesis of the microscopic changes observed in these organs isunknown. There were no histological effects on the spermatogenesis in the testes. Histological

findings were observed in the prostate and epididymides of rat no. 37 suspected of reducedfertility; although a test-item effect on sperm motility could not be excluded, correlation betweenthe observed changes and infertility of male no. 37 was not apparent as all other males from the

high-dose group, similarly affected, were fertile.

All findings were partially (epididymides) or fully (prostate) reversible following a 20 d treatment free period and thus were considered not to be adverse. 

 

The repeated oral administration of the test item in Han Wistar rats over a period of 42 days in males and a minimum of five weeks in females at the doses of 100, 300 and 1000 mg/kg bw/day induced:

From 100 mg/kg bw/day,

- Epithelial vacuolation and decreased secretory content in the prostate (ventral lobe)

From 300 mg/kg bw/day,

- Epithelial vacuolation (caput-body junction) and/or edema/decreased tubular size (cauda) in the epididymides

- Gastric (forestomach) epithelial hyperplasia/hyperkeratosis (indicative of contact irritancy) in males

- Liver centrilobular hepatocellular hypertrophy in males

At 1000 mg/kg bw/day,

- Gastric (forestomach) epithelial hyperplasia/hyperkeratosis in females

Following a 20-day treatment free period, all findings were partially (epididymides and stomach) or fully (prostate and liver) reversible.

 

Reproduction and Development

No effects on reproduction or development were observed in any group.

Mating performance, fertility, duration of gestation, corpora lutea count, implantation rate, post implantation and post natal losses as well as litter size were not affected by the treatment in anygroup.

No test item-related abnormal findings were noted in pups at first litter check or during lactation in any group. Sex ratios at first litter check were unaffected by the exposure to the test item. Pup body weights and body weight gain in dose groups were similar to those in the control group. No test item-related macroscopical findings were noted during necropsy of pups in any group.

 

Conclusion

Based on these results the NOAEL for general toxicity was established at 1000 mg/kg bw/d.

The NOEL for reproduction/developmental toxicity was considered to be 1000 mg/kg bw/d.