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Toxicological information

Toxicity to reproduction

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Administrative data

Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 12 Jan, 2010 to 19 May, 2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2011
Report date:
2011

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
GLP compliance:
yes
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
2-(hydroxymethyl)-2-[[(1-oxoallyl)oxy]methyl]-1,3-propanediyl diacrylate
EC Number:
222-540-8
EC Name:
2-(hydroxymethyl)-2-[[(1-oxoallyl)oxy]methyl]-1,3-propanediyl diacrylate
Cas Number:
3524-68-3
Molecular formula:
C14H18O7
IUPAC Name:
3-(acryloyloxy)-2-[(acryloyloxy)methyl]-2-(hydroxymethyl)propyl acrylate
Constituent 2
Reference substance name:
2-Propenoic acid, reaction product with pentaerythritol
IUPAC Name:
2-Propenoic acid, reaction product with pentaerythritol
Constituent 3
Reference substance name:
2-Propenoic acid, reaction products with pentaerythritol
EC Number:
629-850-6
Cas Number:
1245638-61-2
Molecular formula:
Not available for this UVCB.
IUPAC Name:
2-Propenoic acid, reaction products with pentaerythritol
Details on test material:
- Name of test material (as cited in study report): PETIA
- Physical state: Clear colourless viscous liquid
- Lot/batch No.: JBGK0072T
- Expiration date of the lot/batch: 10 June 2010

Test animals

Species:
rat
Strain:
other: Crl:CD(SD)]
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Inc., Raleigh, NC, USA
- Age at study initiation: ca. 63 d
- Housing: stainless steel wire mesh cages suspended above cage board
- Diet (e.g. ad libitum): PMI Nutrition International, LLC Certified Rodent LabDiet® 5002
- Water (e.g. ad libitum): yes
- Acclimation period: 17 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.8 - 22.7°C
- Humidity (%): 38.2 - 45.1%
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 29 January 2010 To: 19 May 2010

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:

VEHICLE
- Justification for use and choice of vehicle (if other than water): corn oil
- Concentration in vehicle: 0, 5, 15 or 40 mg/ml
- Amount of vehicle (if gavage): 5ml/kg bw
- Lot/batch no. (if required): YF0793, YR1134, and YJ0917
Details on mating procedure:
The animals were paired on a 1:1 basis within each treatment group following 14 d of treatment for the males and females. A breeding record containing the male and female identification numbers and the start date of cohabitation was maintained. Each female was housed in the home cage of the male. Positive evidence of mating was confirmed by the presence of a vaginal copulatory plug or the presence of sperm following a vaginal lavage and verified by a second biologist. Each mating pair was examined daily. The day when evidence of mating was identified was termed gestation day 0. If evidence of copulation was not detected after 14 days of pairing, any females that had not shown evidence of mating were placed in plastic maternity cages.

For the purpose of calculating pre-coital intervals, rats paired over a 12-hour dark cycle were considered to have been paired for 1 day.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Fifteen-day room temperature resuspension homogeneity and stability of the test substance formulated in the vehicle at concentrations of 10 and 200 mg/mL were established in a previous study (Stump, Draft, WIL-738003). Therefore, resuspension homogeneity and stability were not assessed for the 15 and 40 mg/mL formulations prepared for the current study.

Prior to the initiation of dose administration, quadruplicate samples for homogeneity and concentration determination were collected from the top, middle, and bottom strata of the 5 mg/mL non-dosing formulation. However, because the analytical results of the initial samples as well as the back-up samples did not meet the WIL Research Laboratories, LLC’s SOP requirements, a new 5 mg/mL non-dosing formulation was prepared. Quadruplicate samples for homogeneity and concentration determination were collected from the top, middle, and bottom strata of the new 5 mg/mL non-dosing formulation. In addition, quadruplicate samples for resuspension homogeneity and stability determinations were collected from aliquots prepared from this same non-dosing formulation following room temperature storage for 5 and 13 days; the aliquots were stirred for at least 60 minutes prior to sampling. Quadruplicate samples for homogeneity and concentration analyses were collected from the top, middle, and bottom of the test substance formulations prepared for the first week of dose administration; samples were also collected from the middle stratum of the vehicle control formulation. Additionally, quadruplicate samples for concentration analysis were collected from the middle stratum of each dosing formulation and vehicle control formulation prepared for the remainder of the study. One set of duplicate samples from each collection was subjected to the appropriate analyses. The remaining set of duplicate samples was stored frozen (approximately -70°C ± 5°C) as back-up. All analyses were conducted by the Analytical Chemistry Department at WIL Research Laboratories, LLC using a validated high performance liquid chromatography method with ultraviolet absorbance detection.
Duration of treatment / exposure:
The males were dosed once daily during study Days 0-27 (14 days prior to pairing through 1 day prior to scheduled euthanasia) for a total of 28 doses. The females were dosed once daily during study Days 0 through the day prior to euthanasia (14 days prior to pairing through lactation day 4) for a total of 40-47 doses. Females that failed to deliver were dosed through the day prior to euthanasia (post-mating Day 25) for a total of 41 doses.
Frequency of treatment:
Daily
Doses / concentrationsopen allclose all
Dose / conc.:
25 mg/kg bw/day (nominal)
Dose / conc.:
75 mg/kg bw/day (nominal)
Dose / conc.:
200 mg/kg bw/day (nominal)
No. of animals per sex per dose:
12 males and 12 females
Control animals:
yes
Positive control:
None

Examinations

Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: All rats were observed twice daily, once in the morning and once in the afternoon, for moribundity and mortality. A detailed physical examination was conducted weekly on each animal beginning approximately 1 week prior to the initiation of dose administration, including on the day of necropsy.

BODY WEIGHT: Yes
- Time schedule for examinations: Individual male body weights were recorded beginning approximately 1 week prior to the initiation of dose administration, on the first day of dose administration and weekly thereafter until the day prior to scheduled euthanasia. Individual female body weights were recorded beginning approximately 1 week prior to the initiation of dose administration, on the first day of dose administration and weekly thereafter until the day evidence of copulation was observed.

FOOD CONSUMPTION:
- Individual food consumption was recorded on the corresponding weekly body weight days until pairing. Food intake was not recorded during the mating period. Once evidence of mating was observed, female food consumption was recorded on gestation days 0, 4, 7, 11, 14, 17, and 20 and on lactation days 1 and 4. Following mating, food consumption for the female with no evidence of mating was measured on a weekly basis until parturition. When food consumption could not be determined for an animal during a given interval (due to an unscheduled death, as females enter gestation/lactation, weighing error, food spillage, obvious erroneous value, etc.), group mean values were calculated for that interval using the available data.

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at necropsy (Study Day 28 for males and Lactation Day 5 for females)
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes
- How many animals: 6 animals/sex/group

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at necropsy (Study Day 28 for males and Lactation Day 5 for females)
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes
- How many animals: 6 animals/sex/group

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: Yes
- FOB assessments were recorded for 6 animals/sex/group prior to dose administration and fasting for clinical pathology sampling on study day 27 (males) and lactation day 4 (females).
- Locomotor activity counts were recorded for 6 animals/sex/group prior to dose administration on study day 27 (males) and on lactation day 4 (females); the same animals evaluated for FOB were selected for locomotor activity assessment.

GROSS PATHOLOGY

HISTOPATHOLOGY
Litter observations:
LITTER VIABILITY AND DEATHS
Each litter was examined daily for survival, and all deaths were recorded. All pups were individually identified by application of tattoo markings on the digits following completion of parturition. A daily record of litter size was maintained. Intact offspring dying were necropsied using a fresh dissection technique, which included examination of the heart and major vessels (Stuckhardt and Poppe, 1984). The carcass of each pup was then discarded.

LITTER CLINICAL OBSERVATIONS
Litters were examined daily for survival and any adverse changes in appearance or behavior. Each pup received a detailed physical examination on PND 1 and 4. Any abnormalities in nursing behavior were recorded.

LITTER BODYWEIGHTS
Pups were individually weighed on PND 1 and 4. Mean pup weights were presented by sex for each litter and by dose group. When body weights could not be determined for a pup during a given interval (due to an unscheduled death, weighing error, etc.), group mean values were calculated for that interval using the available data. The time periods a given pup was not weighed were left blank or designated as “NA” on the individual report tables.

LITTER SEX DETERMINATION
Pups were individually sexed on PND 0 (if possible), 1, and 4.

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
effects observed, treatment-related
Dermal irritation (if dermal study):
effects observed, treatment-related
Mortality:
mortality observed, treatment-related
Description (incidence):
Test substance-related mortality and/or moribundity occured at 75 and 200 mg/kg bw/day.Test substance-related mortality and/or moribundity occured at 75 and 200 mg/kg bw/day.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
In the 200 mg/kg bw/day group males, lower mean body weight gains were noted during the overall pre-mating (Days 0-13) and treatment (Days 0-27) periods, with correspondingly lower mean food consumption during the pre-mating period. As a result, mean male body weight in this group was 8.4% lower than controls on Day 27. For the 75 mg/kg bw/day group males, slightly lower mean body weight gains were noted throughout the treatment period, with the most severe reduction occurring during Days 21-27. However, the magnitude of these differences was not sufficient to affect mean body weights when compared to controls and mean food consumption was similar to that of controls during the pre-mating period (Days 0-13). The effects were therefore not considered to be treatment-related. Mean body weights, body weight gains and food consumption were unaffected by test substance administration in the 25 mg/kg bw/day group males throughout the study and in the 25, 75, and 200 mg/kg bw/day group females during the pre-mating, gestation and lactation periods.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
See above
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
At the scheduled necropsy, there were higher mean absolute neutrophil counts at 75 and 200 mg/kg bw/day, consistent with the test substance-related ulceration and associated inflammation in the non-glandular stomach.
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
no effects observed
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):
Test substance-related histopathologic alterations included ulceration, epithelial hyperplasia and hyperkeratosis, and/or chronic-active inflammation in the non-glandular stomach at 75 and 200 mg/kg bw/day.
Hypertrophy of the zona fasciculate of the adrenal cortex and lymphoid depletion was also noted in the 200 mg/kg bw/day group males and females, respectively. Mean numbers of corpora lutea, unaccounted-for sites and implantation were similar to control values.
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

Details on results (P0)

Male and female mating and fertility, male copulation and female conception indices, mean number of days between pairing and coitus, gestation length, and the process of parturition were unaffected by test substance administration at all dosage levels. Mean numbers of pups born, live litter size, percentage of males at birth, mean pup body weights, and pup body weight gains were unaffected by parental test substance administration. No test substance-related clinical findings or macroscopic findings for pups that were found dead were noted at any dosage level.

Effect levels (P0)

open allclose all
Key result
Dose descriptor:
NOAEL
Effect level:
> 200 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
reproductive function (sperm measures)
reproductive performance
Key result
Dose descriptor:
NOAEL
Effect level:
75 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain

Results: F1 generation

General toxicity (F1)

Clinical signs:
no effects observed
Mortality / viability:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
not examined
Histopathological findings:
not examined
Other effects:
not examined

Developmental neurotoxicity (F1)

Behaviour (functional findings):
not examined

Developmental immunotoxicity (F1)

Developmental immunotoxicity:
not examined

Details on results (F1)

Mean numbers of pups born, live litter size, percentage of males at birth, mean pup body weights, and pup body weight gains were unaffected by parental test substance administration. No test substance-related clinical findings or macroscopic findings for pups that were found dead were noted at any dosage level.

Effect levels (F1)

Key result
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
> 200 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
mortality
body weight and weight gain

Overall reproductive toxicity

Key result
Reproductive effects observed:
no

Any other information on results incl. tables

None

Applicant's summary and conclusion

Conclusions:
Under the study conditions, the test substance NOAEL for reproductive toxicity and neonatal toxicity was determined to be >200 mg/kg bw/day (highest tested dose).
Executive summary:

A study was conducted to determine the repeated dose and reproduction/developmental toxicity of the test substance, PETIA to rat in a screening study conducted according to OECD Guideline 422 and EPA Guideline OPPTS 870.3650, in compliance with GLP. The test substance was administered orally in corn oil by gavage once daily to 3 groups of 12 male and 12 female Crl:CD(SD) rats. Dosage levels were 25, 75 and 200 mg/kg bw/day, administered at 5 mL/kg. Test substance-related mortality and/or moribundity occured at 75 and 200 mg/kg bw/day. Clinical findings were recorded at all dose levels and included salivation or evidence thereof, yellow and red material primarily around the mouth and wiping of mouth in the bedding (females only). Incidences of rales were also noted sporadically at 75 and 200 mg/kg bw/day. All findings were primarily noted at the time of dose administration or shortly thereafter and were attributed to the irritating properties of the substance. In the 200 mg/kg bw/day group males, lower mean body weight gains were noted during the overall pre-mating (Days 0-13) and treatment (Days 0-27) periods, with correspondingly lower mean food consumption during the pre-mating period. As a result, mean male body weight in this group was 8.4% lower than controls on Day 27. For the 75 mg/kg bw/day group males, slightly lower mean body weight gains were noted throughout the treatment period, with the most severe reduction occurring during Days 21-27. However, the magnitude of these differences was not sufficient to affect mean body weights when compared to controls and mean food consumption was similar to that of controls during the pre-mating period (Days 0-13). The effects were therefore not considered to be treatment-related. Mean body weights, body weight gains and food consumption were unaffected by test substance administration in the 25 mg/kg bw/day group males throughout the study and in the 25, 75, and 200 mg/kg bw/day group females during the pre-mating, gestation and lactation periods. No test substance-related effects were noted during the functional observation battery (FOB) or locomotor activity evaluations at any dosage level. At the scheduled necropsy, there were higher mean absolute neutrophil counts at 75 and 200 mg/kg bw/day, consistent with the test substance-related ulceration and associated inflammation in the non-glandular stomach. Gross observations consisted of thickened non-glandular stomach in the 75 and 200 mg/kg bw/day group males and females and adhesions to the liver or spleen in males at 200 mg/kg bw/day. Test substance-related histopathologic alterations included ulceration, epithelial hyperplasia and hyperkeratosis, and/or chronic-active inflammation in the non-glandular stomach at 75 and 200 mg/kg bw/day. Organ weight changes included higher mean adrenal gland weights in the 75 mg/kg bw/day group males and 200 mg/kg bw/day group males and females and lower mean thymus weight in the 200 mg/kg bw/day group females. Hypertrophy of the zona fasciculata of the adrenal cortex and lymphoid depletion was also noted in the 200 mg/kg bw/day group males and females, respectively. Mean numbers of corpora lutea, unaccounted-for sites and implantation were similar to control values. Male and female mating and fertility, male copulation and female conception indices, mean number of days between pairing and coitus, gestation length and the process of parturition were unaffected by test substance administration at all dosage levels. Mean numbers of pups born, live litter size, percentage of males at birth, mean pup body weights and pup body weight gains were unaffected by parental test substance administration. No test substance-related clinical findings or macroscopic findings for pups that were found dead were noted at any dosage level. Under the study conditions, the test substance NOAEL for reproductive toxicity and neonatal toxicity was determined to be >200 mg/kg bw/day (highest tested dose) (Stump, 2011).