Registration Dossier

Diss Factsheets

Administrative data

Endpoint:
screening for reproductive / developmental toxicity
Remarks:
based on test type (migrated information)
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 guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Combined Repeated Dose and Reproductive / Developmental Toxicity Screening Test (Precursor Protocol of GL 422)
Version / remarks:
adopted 22nd March 1996
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: OPPTS 870.3650, July 2000
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Limit test:
no

Test material

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)
Sex:
male/female
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
Details on mating procedure:
- M/F ratio per cage: 1:1
- Length of cohabitation: max. 14 d
- Proof of pregnancy: vaginal plug + sperm in vaginal smear referred to as day 0 of pregnancy
- After 14 days of unsuccessful pairing replacement of first male by another male with proven fertility.
- Further matings after two unsuccessful attempts: no
- After successful mating each pregnant female was caged (how): individually
Animals in recovery groups were not mated.
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
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
Details on study schedule:
- F1 parental animals not mated (screening study)
- Parturition: All dams were allowed to give birth and rear their litters (F1 pups) up to day 4 post partum. Day 0 post partum was designated as the day on which a female had delivered all her pups.
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; not mated)
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)
Positive control:
no

Examinations

Parental animals: Observations and 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.

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

OTHER:
HAEMATOLOGY: Yes
- Time schedule for collection of blood: main groups: day 14 of the pre-pairing period / recovery groups: at the end of the treatment and at the end of recovery periods
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes, approx. 18 h, water ad libitum
- How many animals: 5 males and 5 females from each main group / all males and females in the recovery groups
- Parameters checked in table 1 were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: main groups: day 14 of the pre-pairing period / recovery groups: at the end of the treatment and at the end of recovery periods
- Animals fasted: Yes, approx. 18 h, water ad libitum
- How many animals: 5 males and 5 females from each main group / all males and females in the recovery groups
- Parameters checked in table 1 were examined.

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: Five males shortly before the scheduled sacrifice / five females on day 3 post partum
- Dose groups that were examined: all dose groups, except recovery groups (as no test item-related effects were detected in animals from main groups)
- Battery of functions tested:
- Cage-side observations: faeces-balls, urine and posture as well as resistance to removal.
- Hand-held observations: muscle tone, constitution, skin, pupil size, palpebral closure, lacrimation, salivation, reaction to handling and general abnormalities.
- Open field observations: level of ambulatory activity including rearing (one minute evaluation), unusual body movements (e.g. spasms, convulsions), gait evaluation, behavior, hair coat, respiration, quantity of faeces-balls and urine.
- Reflexes: blinking, palpebral closure, pinna reflex, extensor thrust response, paw pinch, responsiveness to sharp noise, righting reflex and hearing ability (Preyer’s reflex).
- Measurements / Counts: hind limb / fore limb grip strength.
Oestrous cyclicity (parental animals):
not examined
Sperm parameters (parental animals):
Parameters examined in male parental generation: stages of spermatogenesis and histopathology of interstitial cell structure
Litter observations:
STANDARDISATION OF LITTERS
no (screening study)

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

GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities; possible cause of death was/was not determined for pups born or found dead.
Postmortem examinations (parental animals):
SACRIFICE
Males were sacrificed on the day after completion of the treatment when they were no longer necessary for the assessment of reproductive performance.
Dams 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.

GROSS PATHOLOGY: Yes (see table 1)

HISTOPATHOLOGY: Yes (see table 1)
Postmortem examinations (offspring):
SACRIFICE
- Pups surviving to planned termination were killed by decapitation on Day 4 of lactation.
- These animals were subjected to postmortem examinations (macroscopic examination) as follows:

GROSS NECROPSY
- Gross necropsy consisted of external examinations

HISTOPATHOLOGY / ORGAN WEIGTHS
Not performed
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.
Reproductive indices:
Percentage mating = ( Females mated / Females paired) * 100
Fertility index = ( Females achieving a pregnancy / Females paired) * 100
Conception rate = ( Females achieving a pregnancy / Females mated) * 100
Gestation index = ( Number of females with living pups / Females pregnant) * 100
Offspring viability indices:
Birth index = (pups born alive / number of implantations) * 100
Viability index = (pups alive before culling on day 4 p.p. / pups born alive) * 100

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
effects observed, treatment-related
Body weight and weight changes:
effects observed, treatment-related
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
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

Details on results (P0)

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
Main groups
No test item-related deaths were noted during the study at any dose level.
One male (no. 43) and one female (no. 87) in group 4 as well as one female (no. 84) in group 3 were found dead prior to schedule. All remaining animals survived the scheduled study period. Male no. 43 was found dead on day 6 of the after pairing period, female no. 87 was found dead on day 1 of the lactation period and female no. 84 was found dead on the day of scheduled necropsy, day 4 of the lactation period. No signs of toxicity were observed in these animals. Based on the macroscopical and histopathological findings, an accidental administration error was considered to be the cause of death of the two animals in group 4. No cause of death was identified for the female in group 3.

Cage Side Observations
No test item-related clinical signs were noted in males or females in any group.
The only clinical signs recorded during the study were breathing noises in one male in group 4 (no. 47) observed on five days during the pairing period and in one female in group 4 (no. 96) on one day during the gestation period. Further, hair loss was noted in one female in the control group (no. 53). Due to incidental occurrence in individual animals, these observations were considered not to be related to the treatment with the test item.

Detailed Clinical Observations
No test item-related findings were noted in males or females in any group during detailed weekly
observations. The only finding recorded during the examination was hair loss confirmed in female no. 53.

Recovery groups
- described in detail in IUCLID section “Repeated dose toxicity”

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
Main groups
Treatment with the test item caused a reduction in body weights and body weight gain in group 4. Mean body weight gain in order of ascending dose levels was 4.6%, 4.4%, 4.1% and 2.9% during the pre-pairing period, 6.2%, 5.2%, 5.3% and 3.5% during the pairing period and 2.8%, 4.3%, 3.7% and 2.7% during the after pairing period until day 14.
In group 4, a body weight loss by 0.2% was noted on day 2 of the pre-pairing period followed by slight but statistically significant reduction in body weight gain recorded on several days during the entire study. Reduction in body weight gain resulted in slightly lower body weights. Mean body weights in the high-dose group were approximately 5% lower than the respective control values toward the end of the study with statistical significance on days 10 and 15 of the after pairing period.
In groups 2 and 3, body weights and body weight gain were similar to the respective values in the control group.
Treatment with the test item caused a reduction in body weights and body weight gain in group 4. Mean body weight gain in order of ascending dose levels was 3.4%, 3.6%, 3.4% and 3.3% during the pre-pairing period, 64.4%, 57.2%, 60.3% and 57.9% during the gestation period, and 4.9%, 2.4%, 2.6% and 4.7% during the lactation period.
In group 4, a minor reduction in body weight gain was noted during the gestation period with a statistical significance on day 11 of this period. No statistically significant changes were noted in the absolute body weights in this group.
In groups 2 and 3, body weights and body weight gain were similar to the respective values in the control group.

Recovery groups
- described in detail in IUCLID section “Repeated dose toxicity”

REPRODUCTIVE FUNCTION: SPERM MEASURES (PARENTAL ANIMALS)
Microscopic examination of the testes, including the qualitative examination of the stages of spermatogenesis and evaluation of the uterus, follicles and corpora lutea in the ovaries revealed no test item-related findings.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
Mating performance and fertility were not affected by the treatment with the test item.
With exception for one female in group 3 (no. 78), all females mated within the first pairing period. Precoital time was unaffected by treatment with the test item. Mean (median) precoital times were 2.1 (2), 3.3 (3), 3.2 (2) and 2.1 (2) days in groups 1, 2, 3 and 4, respectively.
For female no. 78 a second pairing period with another male was initiated after no mating had been observed during the first pairing period. This female mated successfully within one day of the second pairing period. With exception for one female in group 3 (no. 76) and one female in group 4 (no. 85), all mated females were pregnant and gave birth to living pups. Female no. 76 was pregnant but did not give birth; one implantation site was found in this female during
necropsy. Female no. 85 was not pregnant.
Consequently, percentage mating (number of females mated as a percentage of females paired), was 100% in all groups, fertility index (number of females achieving pregnancy as a percentage of females paired) and conception rate (number of females achieving pregnancy as a percentage of females mated) were 100% in groups 1, 2 and 3 and 91.7% in group 4, whereas gestation index (number of females with living pups as a percentage of females pregnant) was 100%, 100%, 91.7% and 100% in groups 1, 2, 3 and 4.
Corpora lutea count was not affected by the trea ztment with the test item. Mean number of corpora lutea per dam was 13.4, 13.3, 13.5 and 12.9 in groups 1, 2, 3 and 4, respectively.
Duration of gestation was unaffected by the exposure to the test item. Mean duration of gestation was 21.7, 21.6, 21.7 and 21.6 days, in order of ascending dose level.
Mean number of implantations per dam and post-implantation loss were considered not to be affected by the treatment with the test item. Mean number of implantations per dam was 13.3, 11.5, 12.4 and 12.0 whereas mean incidence of post-implantation loss per dam was 0.8, 1.0, 2.4 and 0.7 in groups 1, 2, 3 and 4, respectively.
Birth index (number of pups born alive as a percentage of implantations) was 93.8%, 91.3%, 80.9% and 93.9% in order of ascending dose levels.
In group 3, a statistically significantly increased total incidence of post-implantation loss and statistically significantly increased number of litters affected were noted. This resulted in statistically significantly lower birth index (number of pups born alive as percentage of implantation sites) in this group. The total number of implantations lost in this group was 26 (in 10 litters) compared to 10 (in 5 litters) in the control group whereas birth index in this group was
80.9% compared to 93.8% in the control group. In group 4, values of post implantation loss (8 implantation sites in 6 litters) and birth index (93.9%) were similar to the control values and therefore observation in group 3 was considered not to be related to the treatment.

ORGAN WEIGHTS (PARENTAL ANIMALS)
Main groups
Treatment with the test item caused an increase in liver weights in males in groups 2, 3 and 4.
The increase of the absolute liver weights as well as increase in liver weights to body weights ratio and liver weight to brain weight ratio was statistically significant in all dose-groups. Remaining statistically significant differences were considered not to be related to the treatment with the test item. In particular, changes in testes weights recorded in males in group 4 and changes in the liver weight recorded in females in group 4 were considered to be secondary to a reduction in body weights at the high dose level. Statistically significant increase in the weights of these organs relative to body weights was recorded, however, in the absence of significant changes in absolute weights of these organs or their weights relative to the brain weights.
Therefore, higher testes relative to body weights in males and higher liver relative to body weights in females in group 4 were considered not to be related to the treatment.

Recovery groups
- described in detail in IUCLID section “Repeated dose toxicity”

GROSS PATHOLOGY (PARENTAL ANIMALS)
Main groups
Males
Treatment with the test item caused crateriform retractions in males in groups 3 and 4. The total numbers of animals affected were 0, 0, 5 and 9, in order of ascending dose levels.
Incidentally, one male (no. 1) in the control group had red brown discoloration of thymus, one male in group 2 (no. 27) had pelvic dilation, one further (no. 39), in addition to crateriform retractions in the stomach, had foci on the lungs and reddish discoloration of mesenteric lymph nodes. Due to the isolated occurrence in individual animals, these findings were considered not to be related to the treatment. Male no. 43 in group 4 had dark red discoloration of the lungs and clear watery fluid in thoratic body cavities which indicated an administration error as a cause of its death. This male had also dark red foci on thymus.
No further macroscopical findings were noted in males in any group.
Females
No findings, which were considered to be test item-related, were noted in any group. Incidentally, alopecia was noted in one female (no. 53) in the control group. One further female (no. 73) in group 3 had dark red discolored thymus and red brown discolored mandibular lymph nodes.
Female no. 87 in group 4, which was found dead on day 1 post partum, had thickened, dark red uterus, dark red foci on the thymus and discolored mesentheric lymph nodes. These observations were partially due to the birth shortly before but give no clear indication of the cause of the death. No further macroscopical findings were noted in females in any group.

Recovery groups
- described in detail in IUCLID section “Repeated dose toxicity”

HISTOPATHOLOGY (PARENTAL ANIMALS)
Main groups
Histopathological examination revealed test item-related lesions in the stomach in males in groups 3 and 4 and females in group 4, in the epididymides in males in groups 3 and 4, in the prostate in males in groups 2, 3 and 4 and in the liver in males in groups 3 and 4.
In the stomach of males in groups 3 and 4 with dose-related incidence and in females in group 4 epithelial hyperplasia and hyperkeratosis of the non-glandular mucosa (forestomach) were observed. Minimal erosion/ulcer(s) of the non-glandular or glandular mucosa was also recorded in a few affected animals. Epithelial hyperplasia/hyperkeratosis was the histological correlate of the crateriform retractions observed at necropsy on the forestomach mucosa.
In the epididymides, minimal to moderate vacuolation of the epithelium (zonal distribution) was noted in groups 3 and 4 with dose-related incidence and severity and minimal edema and decreased tubular size (cauda epididymis) were noted in group 4. Occasional 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.
In the prostate, minimal to slight vacuolation of the glandular epithelium and decreased secretory content were recorded in groups 2, 3 and 4, the incidence and severity being unrelated to the dose levels. Changes were observed in the tubulo-alveoli of the ventral lobe; when present on sections, the lateral and dorsal lobes were unaffected.
In the liver, centrilobular hepatocellular hypertrophy was recorded in males in groups 3 and 4 with dose-related incidence and severity. The enlarged hepatocytes displayed a “ground glass” appearance.
All other microscopic observations were few and were those commonly recorded as spontaneous findings in the Wistar rat.
Microscopic examination of the testes, including the qualitative examination of the stages of spermatogenesis and evaluation of the uterus, follicles and corpora lutea in the ovaries revealed no test item-related findings.
The reason for reduced fertility in animals which did not produce any offspring was not clearly identified. The reproductive organs of female no. 85 (group 4), mated with male no. 37, were unremarkable. Male no. 37 showed changes in the epididymides (moderate vacuolation of the epithelium at the caput-body junction and minimal edema and decreased tubular size in the cauda) and prostate (slight decreased secretory content). However, similar changes were observed in all other males in group 4, for which no reduced fertility was apparent. In female no. 76 (group 3), mated with male no. 28, there were evidences of pregnancy (presence of implantation site within the uterus); the reason for premature fetal loss was not determined histologically.
In animals dead prior to scheduled necropsy, histopathological lesions indicative of gavage accident as cause of death were found in male no. 43 and female no. 87. The male no. 43 had diffuse subacute pulmonary inflammation, which was the histological correlate of the lung discoloration observed at necropsy. Female no. 87 had fibrino-necrotic tracheitis. The cause of death of female rat no. 84 (group 3) was not was not determined histologically.

Recovery groups
- described in detail in IUCLID section “Repeated dose toxicity”

OTHER FINDINGS (PARENTAL ANIMALS)
- described in detail in IUCLID section “Repeated dose toxicity”

Effect levels (P0)

open allclose all
Dose descriptor:
NOEL
Remarks:
reproduction
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
other: no adverse effects on reproduction were observed in any group
Dose descriptor:
NOAEL
Remarks:
general toxicity
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
other: see 'Remark'

Results: F1 generation

General toxicity (F1)

Clinical signs:
not examined
Mortality / viability:
no mortality observed
Body weight and weight changes:
no effects observed
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Histopathological findings:
not examined

Details on results (F1)

VIABILITY (OFFSPRING)
Litter size at first litter check was not affected by the treatment with the test item at any dose level. Mean number of pups per dam was 12.5, 10.5, 10.0 and 11.3 in groups 1, 2, 3 and 4, respectively.
Mean number of living pups per dam on day of scheduled termination was 12.4, 10.3, 10.0 and 10.0. Viability index (number of pups alive on day 4 post partum as a percentage of pups born alive) was 99.3%, 98.4%, 100.0% and 88.7% in order of ascending dose levels.
In group 4, statistically significantly increased total number of pups lost during lactation and, consequently, statistically significantly reduced viability index (number of pups alive at planned termination as a percentage of pups born alive) were noted. These findings were due to the termination of 11 pups in litter no. 87 after the female was found dead on day 1 post partum. If this litter was excluded from calculations, values similar to the respective controls and remaining within historical control range were noted at the high-dose level: a total number of pups lost was 3 (in 3 litters) compared to one pup lost in the control group, mean number of pups lost per dam of 0.3 compared to 0.1 in the control group and viability index was 97.3% compared to 99.3% in the control group. For these reasons post-natal loss in group 4 was considered not to be affected by the treatment with the test item.

BODY WEIGHT (OFFSPRING)
No effects on pup body weights or body weight gain were noted in any group until day 4 post partum.
Mean body weights of pups was 6.56, 6.30, 6.50 and 6.34 g on day 1 post partum and 9.49, 8.91, 9.65 and 9.19 g on day 4 post partum, whereas body weight gain was 45.51%, 40.66%, 45.95% and 44.92% during this period.

GROSS PATHOLOGY (OFFSPRING)
No test item-related macroscopical findings were noted during necropsy of pups in any group. The only findings noted were dark red discoloration of lungs in one pup in the control group and sore in one pup in group 3.

OTHER FINDINGS (OFFSPRING)
No test item-related abnormal findings were noted in pups at first litter check or during lactation in any group.
The only findings recorded during the first litter check or during lactation were malformation of tail or missing base o tail, wound on lower mandible, localized swelling of the nose, maculate erythema on nose and wound nose, blue discolored left thigh were noted in individual pups.

Sex ratios at first litter check and on day 4 post partum were unaffected by exposure to the test item. The proportion of males at first litter check was 51%, 51%, 50% and 60%, in order of ascending dose level.

Effect levels (F1)

Dose descriptor:
NOEL
Remarks:
development
Generation:
F1
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
other: no adverse effects on development were observed in any group

Overall reproductive toxicity

Reproductive 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.