Soybean oil, epoxidized, acrylate

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Remarks:

based on test type (migrated information)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

November 2012-February 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study has been conducted according to OECD Guideline No. 422 and under GLP.

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 (incl. QA statement)

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH, Sandhofer Weg 7, D-97633
- Age at study initiation: young adult rats, approximately 10 weeks old at starting and 12 weeks at mating
- Weight at study initiation: Males: 346 g – 407 g, Females: 224 g - 277 g
- Housing: Type II and/or III polypropylene/polycarbonate
- Diet (e.g. ad libitum): ssniff® SM R/M-Z+H "Autoclavable complete feed for rats and mice – breeding and Maintenance" produced by ssniff Spezialdiäten GmbH, D-59494 Soest Germany ad libitum,
- Water (e.g. ad libitum): tap water from municipal supply, as for human consumption from 500 ml bottle ad libitum.
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.4.– 23.1°C
- Humidity (%): 32 - 57%
- Air changes (per hr): 15-20 air exchanges/hour
- Photoperiod (hrs dark / hrs light): 12 hours daily, from 6.00 a.m. to 6.00 p.m.

IN-LIFE DATES: From: 12 November 2012 To: 30 December 2012

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
The test item was formulated in Poly(ethylene glycol) 400 at 15.6, 62.5 and 250 mg/mL in the Central Dispensary of CiToxLAB Hungary Ltd. Formulations were prepared fresh prior to administration to animals.

Details on mating procedure:

Mating began after the animals have attained full sexual maturity, 2 weeks after the initiation of treatment, with one female and one male of the same dose group (1:1 mating) in a single cage. Females remained with the same male until copulation occurred, for up to 5 days. A vaginal smear were prepared daily during the mating period and stained with 1% aqueous methylene blue solution. The smears were examined with a light microscope, the presence of vaginal plug or sperm in the vaginal smear was considered as evidence of copulation (Day 0 of pregnancy as defined by the relevant guidelines). Sperm positive females were caged individually. Mating pairs were clearly identified in the data, mating of siblings was avoided.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Test item formulations were analysed for concentration and homogeneity at Analytical Laboratory of CiToxLAB Hungary Ltd. Top, middle and bottom triplicate samples were taken from test item formulations on 4 occasions, during the first, second and last weeks and approximately midway of the treatment. Two sets to analyse (which were collected in replicates as practical) and one set as a back-up for any confirmatory analyses. Similarly, one sample was taken in duplicate from the vehicle control group 1 solution for concentration measurements.

Duration of treatment / exposure:

Test item or negative control material treated groups animals were administered the dosing solutions daily on a 7 days/week basis, by oral gavage using a tipped gavage needle attached to a syringe. A constant volume was administered to all animals. The actual volume administered was calculated and adjusted based on each animal’s most recent body weight.

Details on study schedule:

The dose levels and the vehicle were selected based on available data, formulation and analytical trials and information from previous experimental work, including the results of a repeated dose range finding study in the rat (CiToxLAB Hungary Ltd. study code 12/074-220PE), with the aim of inducing toxic effects but ideally no death or suffering at the highest dose and a NOAEL at the lowest dose. The oral route was selected as it is a possible route of exposure to the test item in humans.

Test item or negative control material treated groups animals were administered the dosing solutions daily on a 7 days/week basis, by oral gavage using a tipped gavage needle attached to a syringe. A constant volume was administered to all animals. The actual volume administered was calculated and adjusted based on each animal’s most recent body weight.

Dosing of both sexes began after an acclimation period (A) of 6 days after the animal arrival; the animals were dosed for 2 weeks before mating, during the mating/post-mating, and were continued up to the day of necropsy.

Males were dosed for 28 days (14 days pre-mating and 14 days mating/post-mating period), then were euthanized and subjected to necropsy examination, as no additional mating was considered required. Females were dosed for 14 days pre-mating, for up to 5 days mating period, through gestation and up to the day of necropsy (at least 4 days post-partum dosing). In one female at 1000 mg/kg (no. 4506) the duration of mating period was 9 days, however this female proved to be not pregnant. The day of birth (viz. when parturition was complete) was defined as Day 0 post-partum. Females showing no-evidence of copulation were sacrificed, 25-27 days after the end of the mating period.

Mating began after the animals have attained full sexual maturity, 2 weeks after the initiation of treatment, with one female and one male of the same dose group (1:1 mating) in a single cage. Females remained with the same male until copulation occurred, for up to 5 days. A vaginal smear were prepared daily during the mating period and stained with 1% aqueous methylene blue solution. The smears were examined with a light microscope, the presence of vaginal plug or sperm in the vaginal smear was considered as evidence of copulation (Day 0 of pregnancy as defined by the relevant guidelines). Sperm positive females were caged individually. Mating pairs were clearly identified in the data, mating of siblings was avoided.

All F1 offspring were terminated on Day 4 post-partum; in order to allow for overnight fasting of dams prior to urine collection on PPD5, offspring were euthanized on PND/PPD 4, and the dams on PPD/PND 5.

Remarks:

Doses / Concentrations:
62.5, 250 and 1000 mg/kg bw
Basis:
nominal conc.

No. of animals per sex per dose:

12 animals/sex/dose

Control animals:

yes, concurrent vehicle

Details on study design:

DOSE SELECTION RATIONALE
The dose levels and the vehicle were selected based on available data, formulation and analytical trials and information from previous experimental work, including the results of a repeated dose range finding study in the rat (CiToxLAB Hungary Ltd. study code 12/074-220PE), with the aim of inducing toxic effects but ideally no death or suffering at the highest dose and a NOAEL at the lowest dose. The oral route was selected as it is a possible route of exposure to the test item in humans.

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS:
All animals: Animals were inspected for signs of morbidity and mortality twice daily, at the beginning and end of the working day. General clinical observations were performed daily, after treatment at approximately the same time with minor variations, or in the afternoon (pm) as practical during the working day, as no peak period of effects was noted after dosing during the first days of treatment. All animals were monitored for pertinent behavioural changes, signs of difficult or prolonged parturition and all signs of toxicity including mortality. Any changes were recorded including onset, degree and duration of signs as applicable.

DETAILED CLINICAL OBSERVATIONS:
More detailed examinations were made once before the first exposure (to allow for within-subject comparisons), then at least weekly, in the morning (am) or before treatment. These observations were made outside the home cage in a standard arena, at similar times as practical. The animals were monitored for changes in skin, fur, eyes, mucous membranes, occurrence of secretions and excretions, and autonomic activity (e.g. lachrymation, piloerection, pupil size, unusual respiratory pattern), or changes in gait, posture and response to handling as well as the presence of clonic or tonic movements, stereotypies (e.g. excessive grooming, repetitive circling), difficult or prolonged parturition or bizarre behaviour (e.g. self-mutilation, walking backwards); special attention were directed towards the observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma. No such clinical signs were observed during the study.

NEUROBEHAVIOURAL EXAMINATION
5 males and 5 females/group, “subgroup A”: Assessment of any potential test item related neurotoxicity was performed during the last exposure week (males on Day 24am; females on PPD 3am). In order to avoid hyperthermia of pups, dams were removed from the pups for not more than approximately 30-40 minutes. Selected animal were subjected to the functional observation battery, including qualitative assessment of the grip strength, and to measurements of the landing foot splay and fore/hind grip strength.

To measure the landing foot splay, the fore/hind paws of the rat were painted with ink and the rats were dropped from a horizontal position onto the appropriate record sheet covering the examination table. The distance between the two resulting ink spots was measured.

Fore/hind grip strength measurements were conducted using a grip strength meter (Model GS3, Bioseb, Chaville, France), an instrument designed to quantify objectively rodent muscular strength, in order to identify and assess quantitatively any potential effect of test item. The rats were held appropriately such that the fore limbs were allowed to grip the support bar and gently pulled back until they released the bar; the device measured the maximum grip strength. This was performed 3 times for each animal on each test day. The procedure was repeated with the hind limbs with the appropriate grip support; results are tabulated with individual and mean data.

Sensory reactivity to different type of stimuli (e.g. auditory, visual and proprioceptive), assessment of grip strength and motor activity were conducted and the general physical condition and behaviour of animals were tested. A modified Irwin test was performed. Parameters such as, but not limited to body position, locomotor activity, respiration rate, respiration type, piloerection, head searching compulsive biting or licking, circling, upright walking, retropulsion, jumping, exophthalmos, twitches, clonic convulsions, tonic convulsions, tremor, startle, transfer arousal, spatial locomotion, gait, posture, limb position, finger approach, finger withdrawal, touch escape response, diarrhoea, diuresis, visual placing, grip strength, body tone, corneal reflex, pinna, toe pinch, grasping reflex, positional struggle, skin, mucous membrane colour, salivation, palpebral closure, lachrymation, limb tone, abdominal tone, tail pinch, righting reflex, and/or vocalisation were evaluated.

BODY WEIGHT:
All adult animals were weighed with accuracy of 1 g for randomization purposes, then on Day 0, afterwards at least weekly and at termination. Parent females were weighed on gestation Days GD 0, 7, 14 and 20 and on postpartal Days PPD0 (within 24 hours after parturition), PPD4 and PPD5 (before termination). Body weights of the female animals were additionally weighed on gestational Days GD10 and 17 in order to give accurate treatment volumes, but these data were not evaluated statistically.

FOOD CONSUMPTION
Animal food consumption was determined by re-weighing the non-consumed diet with a precision of 1 g on Day 7 then at least weekly (see Study Schedule).

OPHTHALMOSCOPIC EXAMINATION:
The fundus of eyes of all animals was examined before treatment. Five male and 5 female Control and High dose animals (“subgroup A”) randomly selected from groups 1 and 4, during the last week of treatment prior to necropsy (males, Day 24pm, females, PPD 3 pm). Mydriasis was produced after instillation of a mydriatic agent (eye drops "Humapent") into the conjunctival sac. The examination was performed using a Gowlland ophthalmoscope. As no ophthalmoscopic alterations were found, no additional examination was performed in other animals.

OBSERVATION OF THE DELIVERY PROCESS, OFFSPRING AND NURSING INSTINCT
Females were allowed to litter and rear their offspring. Delivery process was observed as carefully as possible. All observations were recorded as applicable. No evidence of abnormal deliveries was recorded. The duration of gestation was recorded and was calculated from Day 0 of pregnancy. Dams were observed to record whether they form a nest from the bedding material and cover their new-borns or not. The efficiency of suckling was observed by the presence of milk in the pups' stomach. All observations were recorded as applicable.

Each litter was examined as soon as possible after delivery to establish the number and sex of pups, stillbirths, live births, runts (pups that are significantly smaller than normal pups) and the presence of gross abnormalities.

Observations are reported individually for each adult animal. In addition to the observations on parent animals, the pups (offspring) were monitored for any behavioural changes. Live pups were counted, sexed, weighed individually within 24 hours of parturition (ex. Day 0 or 1 post-natal, PND0 or 1) and on PND4, with accuracy of 0.01g. All the litters were checked and recorded daily for the number of viable and dead pups. The pups found dead and intact (not cannibalized) were subjected to necropsy with macroscopic examination in order to identify the possible cause of death. All observed abnormalities were recorded

HAEMATOLOGY:
All animals selected for blood sampling were fasted (overnight period of food deprivation). 5 males and 5 females/group, “subgroup B”: For terminal blood sampling of animals selected (subgroup B), 3 samples were taken from each scheduled animal: for haematology (in tubes with K3-EDTA as anticoagulant), one sample for blood clotting times (APTT and PT measurements, in tubes with sodium citrate as anticoagulant) and one to obtain serum (in tubes with no anticoagulant) for clinical chemistry. Blood samples were collected by cardiac puncture under pentobarbital anaesthesia, immediately prior to scheduled necropsy.-Parameters checked in table on page 22 of the report were examined.

CLINICAL CHEMISTRY:
- Time schedule for collection of blood: immediately prior to scheduled necropsy.
- Animals fasted: Yes
- How many animals: 5 males and 5 females
- Parameters checked in table on page 23 of the report were examined.

URINALYSIS:
Urine sampling (approximately 16 hours sampling period) was performed prior to necropsy (males on Day 28; females on PND 5). Parameters checked in table on page 24 of the report were examined.

Litter observations:

LITTER OBSERVATIONS
Females were allowed to litter and rear their offspring. Delivery process was observed as carefully as possible. All observations were recorded as applicable. No evidence of abnormal deliveries was recorded. The duration of gestation was recorded and was calculated from Day 0 of pregnancy. Dams were observed to record whether they form a nest from the bedding material and cover their new-borns or not. The efficiency of suckling was observed by the presence of milk in the pups' stomach. All observations were recorded as applicable.

Each litter was examined as soon as possible after delivery to establish the number and sex of pups, stillbirths, live births, runts (pups that are significantly smaller than normal pups) and the presence of gross abnormalities.

GROSS EXAMINATION OF DEAD PUPS:
Observations are reported individually for each adult animal. In addition to the observations on parent animals, the pups (offspring) were monitored for any behavioural changes.

PARAMETERS EXAMINED
Live pups were counted, sexed, weighed individually within 24 hours of parturition (ex. Day 0 or 1 post-natal, PND0 or 1) and on PND4, with accuracy of 0.01g. All the litters were checked and recorded daily for the number of viable and dead pups. The pups found dead and intact (not cannibalized) were subjected to necropsy with macroscopic examination in order to identify the possible cause of death. All observed abnormalities were recorded.

Postmortem examinations (parental animals):

SACRIFICE
Surviving adult rats were terminated according to the study protocol. These adult animals were euthanized upon completion of the study, according to the study plan.

GROSS NECROPSY
Gross necropsy was performed on all animals. Terminally, after completion of the treatment, animals were euthanised under pentobarbital anaesthesia, followed by exsanguination. After exsanguination the external appearance was examined, cranium, thoracic and abdominal cavities were opened and the appearance of the tissues and organs were observed macroscopically. Any abnormality was recorded with details of the location, colour, shape and size, as appropriate.

HISTOPATHOLOGY / ORGAN WEIGHTS

Postmortem examinations (offspring):

SACRIFICE
- The F1 offspring were sacrificed at 4 days of age.

GROSS NECROPSY
Pups euthanized at PND 4 were carefully examined at least externally for gross abnormalities. Any pups showing abnormalities in structure or behaviour, including the pups found dead and intact (not cannibalized) were subjected to necropsy with macroscopic examination, in order to identify the probable cause of death if possible.

Statistics:

Data were collected using the software PROVANTIS v.7, or were recorded on the appropriate forms from the relevant SOPs of CiToxLAB Hungary Ltd., then tabulated using the software PROVANTIS v.7, Microsoft Office Word and/or Excel, as appropriate. Numerical data obtained during the conduct of the study were subjected as appropriate to calculation of group means and standard deviations.

The statistical evaluation of appropriate data (marked † below) was performed with the statistical program package SPSS PC+4.0. The homogeneity of variance between groups was checked by Bartlett’s homogeneity of variance test. Where no significant heterogeneity was detected, a one-way analysis of variance (ANOVA) was carried out. If the obtained result was significant, Duncan Multiple Range test was used to access the significance of inter-group differences. Getting significant result at Bartlett’s test, the Kruskal-Wallis analysis of variance was used and the inter-group comparisons were performed using Mann-Whitney U-test. Chi2 test was performed as feasible.

Reproductive indices:

In parental males the following parameters were detemined: number of fertile pairings, number of infertile males, male mating index and male fertility index.

In parental females the following parameters were detemined: number of pairings, number of pregnant females, number of sperm positive, but non-pregnant females, number of non mated females, female mating index, female fertility index, gestation index, duration of pregnancy (days), number of corpora lutea/dams, number of implantations/dams, number of dams with live pups (day 0 and 4), pre-implantation mortality, intrauterine mortality,
total mortality (intra and extra uterine mortality).

Offspring viability indices:

In offspring the following parameters were detemined: mean pup body weight (per pup within the group and per litter) on PND 0 and 4, mean pup body weight gain (per litter) between postnatal Days 0-4, number of live births per litter, and number of viable pups per litter on postnatal Days 0 and 4, survival Index of pups on postnatal Days 0 and 4, sex ratio % (on postnatal Days 0 and 4).

Clinical signs:

no effects observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Other effects:

no effects observed

Reproductive function: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

not examined

Reproductive performance:

no effects observed

DOSE FORMULATION ANALYSIS
Test item content and homogeneity of the dosing formulations was determined on 3 occasions during the study. Dose formulations were homogenous. The measured concentrations of Epoxidized Soybean Oil Acrylate evaluated for each test item-dose group varied between 90.7 ± 4.8% and 102.9 ± 3.2%. No test item was detected in the control samples. These results were within acceptable ranges (90 % - 110%).

PARENTAL/ADULT EVALUATION

MORTALITY
There was no unscheduled test item related mortality during the study. One female animal in the control group was found dead on Day 4, considered a technical error during dosing.

CLINICAL SIGNS
No test item-related adverse effects or systemic clinical signs were noted following administration of daily by oral gavage under the conditions of this study, or in the animals. In the male animals, thin fur (in one animal in the low dose group) and liquid faeces (in one animal in the high dose group) were observed. In the female animals scab and scar (in one - one animal in the control, mid and high dose groups), fur thin (in one animal in the low dose group), faeces liquid (in one animal in the control group) and subcutaneous mass (in one animal in the high dose group) were observed occasionally.

NEUROBEHAVIOUR
There were no toxicologically significant changes in the animal behaviour, general physical condition, in the reactions to different type of stimuli, grip strength or motor activity in the control or treated groups, at the evaluation performed towards the end of the treatment period.

Increased vocalization was observed on occasion in the animals (one male and two females in the control group and one female in low dose group) and slightly increased startle (one male in control group) when subjected to the modified Irwin test (functional observation battery). However, no treatment-related differences to the control, or dose, or gender related response, were noted, and this sign was considered to be without toxicological significance and within the normal biological variation with respect to behaviour, reactions to different type of stimuli or manipulations.

Slightly decreased mean grip strength (g) were measured in high dose males attaining statistical difference (p<0.05) for forelimbs and without statistical difference for hindlimbs when compared to control. Increased landing foot splay (cm) values were measured in the High dose male rats with statistical significance (p<0.05) for forelimbs and without statistical significance at hindlimbs.

Based on the lack of a consistent dose or gender response and the normal historical ranges expected, these variations were regarded as incidental and without toxicological significance.

OPHTHALMOSCOPIC EXAMINATION
No test item related changes compared to pre-treatment were noted during ophthalmoscopy examination of 5 male and 5 female control and high dose animals during the last week of treatment prior to necropsy (males, Day 24 pm, females, PPD 3 pm), thus no additional evaluation was required in other dose groups .

BODY WEIGHT AND WEIGHT GAIN
No test item related effects were noted on the mean body weight and body weight gain values following daily administration of Epoxidized Soybean Oil Acrylate at dose levels up to and including 1000 mg/kg bw/day, during the treatment period. A higher increase in body weight gain was recorded for low dose males between days 21 and 27 (p<0.05) and High dose females between days 0 - 7 and 0 – 14 (p<0.05) compared to the control animals. However, all body weight gains in the study were within the normal range and showed no clear dose response relationship. Based on the isolated incidence and lack of a consistent dose or gender response, these variations were regarded as incidental and without toxicological significance.

FOOD CONSUMPTION
There were no test item-related differences in the mean daily food consumption in any test-item treated group (62.5, 250, or 1000 mg/kg bw/day) when compared to the control. Minor differences to control were noted or variations within the group, generally associated with changes in the study schedule including mating, delivery, or fasting before blood collection for clinical pathology evaluation, unrelated to treatment and with no toxicological significance. Occasionally, spillage was noted in the low and mid dose females.

HAEMATOLOGY
When compared to the controls, there were no differences that were considered toxicologically significant in the treated animals. Variations were noted in a few parameters in the female animals, on occasion attaining statistical significance, including for example statistically higher mean cell haemoglobin concentration (MCHC) in the Low dose(p<0.05), lower than control red cell distribution width (RDW) in the Low dose (p<0.05) and higher white blood count (WBC) than control in the Mid dose(p<0.05). Evaluation of the mean and individual results in comparison with the control data did not reveal any test-item related cause of the changes noted, and/or no consistent dose or gender-related response was observed. Therefore, these differences observed between the Control and treated groups were considered to be incidental or individual findings, which were not related to treatment, were generally comparable with the expected physiological range and of no toxicological significance.

CLINICAL CHEMISTRY
In the animals evaluated at the completion at termination (on Day 28 in males and on PPD5 in females), there were no toxicologically significant changes or adverse effects on the animal serum chemistry that could be ascribed to Epoxidized Soybean Oil Acrylate administration in the conditions of this study. A few clinical chemistry parameters showed on occasion statistically significant variations (i.e. albumin concentration 5.5% (p<0.05), calcium 7% (p<0.01) and phosphorus 23% (p<0.01) lower than control in the low dose females, however, there was no dose or gender response or the values were within the physiological ranges. For this reason, these variations were not considered toxicologically significant or related to treatment.

URINALYSIS
Epoxidized Soybean Oil Acrylate administration daily by oral gavage at up to and including 1000 mg/kg bw/day did not result in any test item-related effects considered adverse at urinalysis performed prior to necropsy in the animals. The urine volume showed minor variations, without statistically significant in the animals, however, with no dose or gender-dependent, and not considered to be of toxicological importance in correlation with test item administration in the conditions of this study. The few other minor variations observed did not attain statistical significance and/or were regarded as normal background changes.

OESTRUS CYLCE, REPRODUCTIVE ABILITY ASSESSMENT AND INDICES
There were no statistically significant differences between the control and test item-treated groups with regard to reproductive ability or in the mating or gestation indices, or effects considered adverse or toxicologically significant in correlation with Epoxidized Soybean Oil Acrylate administration.
In all groups, the mating indices were 100%, the fertility indices, 100%, 100%, 92% and 83% due to 0/12, 0/12, 1/12 and 2/12 non-pregnant females, respectively, in Groups 1, 2, 3 and 4 (these values are all in the normal historical range). The gestation indices was 100% values in all treated groups.

Test item administration was considered to have no impact on the duration of the mating period. Successful coitus (sperm positive vaginal smears and/or vaginal plugs) generally occurred within up to 5 days of pairing (cohabitation). In one of 12 female at 1000 mg/kg (no. 4506) the duration of mating period was 9 days, almost only dioestrus picture with leucocytes characterized the oestrus cycle, however this female proved to be not pregnant. This was considered to be an incidental finding previously seen in control females.

EVALUATION OF THE GESTATION, PARTURITION AND POST-PARTAL PERIOD
No test item effect on the duration of pregnancy or abnormalities in the gestation outcome ascribed to the treatment were observed. The mean duration of pregnancy was similar in the control and test item treated groups, all were 22 to 23 days. All the parturitions were normal. The number of corpora lutea and implantation sites were comparable in the treated groups up to and including 1000 mg/kg bw/day with the mean value recorded in the Control group.

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

no effects observed

Sexual maturation:

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings:

not examined

VIABILITY (OFFSPRING)
Epoxidized Soybean Oil Acrylate administered to parental generation at up to 1000 mg/kg bw/day did not lead to mortality or any adverse effects considered related to treatment or toxicologically significant in the F1 generation.

CLINICAL SIGNS (OFFSPRING)
No abnormal behaviour of the pups was noted. No external abnormalities clearly ascribed to treatment were detected at the clinical or external macroscopic examinations of the pups. In single pups at control, 62.5 and 1000 mg/kg haemorrhage or cold body were observed on PND0. There were two animals found (one control and one low dose) with malrotated hind limbs. In the stillborn population there were 3 pups with external alteration, ie. exencephaly (absence of the cranial region of the head, with the brain reduced) in one pup in the low dose; anus absent, short mandible and maxilla, protruding tongue and malpositioned right ovary in one pup in the mid dose; absent tail in one animal in High dose. These 3 abnormalities are changes occasionally seen in control pups/foetuses, so they are not considered to represent a clear effect of treatment. No other external abnormalities were detected at the clinical or external macroscopic examinations of the pups. The incidence of these findings was low, there was no dose response, the types of finding have been seen in control animals in the past in Wistar rats; so it is considered that the observations do not clearly reflect a test item effect.

The number of viable pups on PND4 as well as pups survival indices on PND0 and PND4 when evaluated as litter data were comparable to control values at up to and including 1000 mg/kg bw/day, or showed minor variations ascribed to individual, biological variability.
 
BODY WEIGHT (OFFSPRING)
There were no effects considered adverse on the offspring weight or weight gain following administration of Epoxidized Soybean Oil Acrylate at 62.5, 250 or 1000 mg/kg bw/day to parental generation under the conditions of this study. When evaluated per litter basis, the mean litter body weights and/or body weight gain on PND 0 and 4 showed no statistically or toxicologically significant differences compared to controls in the F1 generation.

Compared to control, statistical significance was noted in the mean body weights values evaluated for all pups at 1000 mg/kg bw/day (high dose) on PND0 (below control) (p<0.05) and PND4 (above control) (p<0.05). Slightly lower mean body weight gain values were measured for 62.5 mg/kg (low dose) (p<0.01) and slightly higher values for 1000 mg/kg bw/day (high dose) between PND0 - 4 (p<0.05 and p<0.01, respectively). All values were in the normal range, no differences were considered to be related to treatment with test item.

GROSS PATHOLOGY (OFFSPRING)
No macroscopic changes were seen in F1 offspring generation euthanized and examined externally at scheduled termination on PND 4.

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Reproductive effects observed:

not specified

Conclusions:

The no observed adverse effect level (NOAEL) for Epoxidized Soybean Oil Acrylate for parental/adult and F1 effects is considered to be 1000 mg/kg bw/day.

Executive summary:

The purpose of this Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test in the Rat study was to obtain information on the toxicity of the test item following repeated daily administration by oral gavage to Wistar rats. The study also included a reproductive/developmental toxicity-screening test, intended to provide initial information on possible effects on male and female reproductive performance such as gonadal function, mating behaviour, conception, pregnancy, parturition and also on the development of the F1 offspring from conception to Day 4 post-partum. The study was conducted according to OECD Guideline No. 422.

Male and female Wistar rats were treated for 2 weeks pre-mating, then during the mating/postmating period, males for 28 days and females throughout gestation period and up to and including postpartum/lactation Day PPD5

Daily administration of Epoxidized Soybean Oil Acrylate by oral gavage to Wistar rats did not result in test item related mortality or clinical adverse effects at daily, weekly or neurological assessment, in ophthalmological changes, or changes in the body weight, food consumption, haematology, coagulation, clinical chemistry, or urinalysis parameters at dose levels of 62.5, 250, or 1000 mg/kg bw/day during the treatment period under the conditions of this study.

 

No test item related, or adverse effects were noted at evaluation of the reproductive parameters during mating and gestation, delivery and post-partum/lactation period until PPD5, under the conditions of this study.There were no adverse effects ascribed to test item administration on the F1 offspring viability, clinical signs, development or at observations following euthanasia. There were no test item-related changes observed in organ weights, at necropsy or at histopathology for the adult animals of either sex.

 

In conclusion, under the conditions of this study, the no observed adverse effect level (NOAEL) for Epoxidized Soybean Oil Acrylate for parental/adult and F1 effects is considered to be 1000 mg/kg bw/day.

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

According to the Regulation EC 1272/2008, a two-generation reproductive toxicity study (one species, male and female, most appropriate route of administration, having regard to the likely route of human exposure) must be proposed in the REACH dossier (Annex IX) if the 28-day or 90-day study indicates adverse effects on reproductive organs or tissues. An OECD 422 study (28 day exposure) via the oral route is available which has not shown any adverse effects, either in the adult animals or in the offspring at the highest dose (1000 mg/kg bw). Therefore, a two-generation reproductive toxicity study is not proposed.

Short description of key information:
No test item related, or adverse effects were noted at evaluation of the reproductive parameters during mating and gestation, delivery and post-partum/lactation period, under the conditions of this study.There were no adverse effects ascribed to test item administration on the F1 offspring viability, clinical signs, development or at observations following euthanasia. There were no test item-related changes observed in organ weights, at necropsy or at histopathology for the adult animals of either sex. The NOAEL for Epoxidized Soybean Oil Acrylate for parental/adult and F1 effects is considered to be 1000 mg/kg bw/day.

Justification for selection of Effect on fertility via oral route:
There is only a screening reproduction/developmental toxicity study available. The study has been conducted according to OECD guideline no. 422 and under GLP.

Effects on developmental toxicity

Description of key information

No test item related, or adverse effects were noted at evaluation of the reproductive parameters during mating and gestation, delivery and post-partum/lactation period, under the conditions of this study.There were no adverse effects ascribed to test item administration on the F1 offspring viability, clinical signs, development or at observations following euthanasia. There were no test item-related changes observed in organ weights, at necropsy or at histopathology for the adult animals of either sex. The NOAEL for Epoxidized Soybean Oil Acrylate for parental/adult and F1 effects is considered to be 1000 mg/kg bw/day.

Link to relevant study records

Reference

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

November 2012-February 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study has been conducted according to OECD Guideline No. 422 and under GLP.

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

other: OECD 422

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Species:

rat

Strain:

Wistar

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH, Sandhofer Weg 7, D-97633
- Age at study initiation: young adult rats, approximately 10 weeks old at starting and 12 weeks at mating
- Weight at study initiation: Males: 346 g – 407 g, Females: 224 g - 277 g
- Housing: Type II and/or III polypropylene/polycarbonate
- Diet (e.g. ad libitum): ssniff® SM R/M-Z+H "Autoclavable complete feed for rats and mice – breeding and Maintenance" produced by ssniff Spezialdiäten GmbH, D-59494 Soest Germany ad libitum,
- Water (e.g. ad libitum): tap water from municipal supply, as for human consumption from 500 ml bottle ad libitum.
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.4.– 23.1°C
- Humidity (%): 32 - 57%
- Air changes (per hr): 15-20 air exchanges/hour
- Photoperiod (hrs dark / hrs light): 12 hours daily, from 6.00 a.m. to 6.00 p.m.

IN-LIFE DATES: From: 12 November 2012 To: 30 December 2012

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Test item formulations were analysed for concentration and homogeneity at Analytical Laboratory of CiToxLAB Hungary Ltd. Top, middle and bottom triplicate samples were taken from test item formulations on 4 occasions, during the first, second and last weeks and approximately midway of the treatment. Two sets to analyse (which were collected in replicates as practical) and one set as a back-up for any confirmatory analyses. Similarly, one sample was taken in duplicate from the vehicle control group 1 solution for concentration measurements.

Details on mating procedure:

Mating began after the animals have attained full sexual maturity, 2 weeks after the initiation of treatment, with one female and one male of the same dose group (1:1 mating) in a single cage. Females remained with the same male until copulation occurred, for up to 5 days. A vaginal smear were prepared daily during the mating period and stained with 1% aqueous methylene blue solution. The smears were examined with a light microscope, the presence of vaginal plug or sperm in the vaginal smear was considered as evidence of copulation (Day 0 of pregnancy as defined by the relevant guidelines). Sperm positive females were caged individually. Mating pairs were clearly identified in the data, mating of siblings was avoided.

Duration of treatment / exposure:

Test item or negative control material treated groups animals were administered the dosing solutions daily on a 7 days/week basis, by oral gavage using a tipped gavage needle attached to a syringe. A constant volume was administered to all animals. The actual volume administered was calculated and adjusted based on each animal’s most recent body weight.

Remarks:

Doses / Concentrations:
62.5, 250 and 1000 mg/kg bw
Basis:
nominal conc.

No. of animals per sex per dose:

12 animals/sex/dose

Details on study design:

The dose levels and the vehicle were selected based on available data, formulation and analytical trials and information from previous experimental work, including the results of a repeated dose range finding study in the rat (CiToxLAB Hungary Ltd. study code 12/074-220PE), with the aim of inducing toxic effects but ideally no death or suffering at the highest dose and a NOAEL at the lowest dose. The oral route was selected as it is a possible route of exposure to the test item in humans.

Test item or negative control material treated groups animals were administered the dosing solutions daily on a 7 days/week basis, by oral gavage using a tipped gavage needle attached to a syringe. A constant volume was administered to all animals. The actual volume administered was calculated and adjusted based on each animal’s most recent body weight.

Dosing of both sexes began after an acclimation period (A) of 6 days after the animal arrival; the animals were dosed for 2 weeks before mating, during the mating/post-mating, and were continued up to the day of necropsy.

Males were dosed for 28 days (14 days pre-mating and 14 days mating/post-mating period), then were euthanized and subjected to necropsy examination, as no additional mating was considered required. Females were dosed for 14 days pre-mating, for up to 5 days mating period, through gestation and up to the day of necropsy (at least 4 days post-partum dosing). In one female at 1000 mg/kg (no. 4506) the duration of mating period was 9 days, however this female proved to be not pregnant. The day of birth (viz. when parturition was complete) was defined as Day 0 post-partum. Females showing no-evidence of copulation were sacrificed, 25-27 days after the end of the mating period.

Mating began after the animals have attained full sexual maturity, 2 weeks after the initiation of treatment, with one female and one male of the same dose group (1:1 mating) in a single cage. Females remained with the same male until copulation occurred, for up to 5 days. A vaginal smear were prepared daily during the mating period and stained with 1% aqueous methylene blue solution. The smears were examined with a light microscope, the presence of vaginal plug or sperm in the vaginal smear was considered as evidence of copulation (Day 0 of pregnancy as defined by the relevant guidelines). Sperm positive females were caged individually. Mating pairs were clearly identified in the data, mating of siblings was avoided.

All F1 offspring were terminated on Day 4 post-partum; in order to allow for overnight fasting of dams prior to urine collection on PPD5, offspring were euthanized on PND/PPD 4, and the dams on PPD/PND 5.

Maternal examinations:

CAGE SIDE OBSERVATIONS:
All animals: Animals were inspected for signs of morbidity and mortality twice daily, at the beginning and end of the working day. General clinical observations were performed daily, after treatment at approximately the same time with minor variations, or in the afternoon (pm) as practical during the working day, as no peak period of effects was noted after dosing during the first days of treatment. All animals were monitored for pertinent behavioural changes, signs of difficult or prolonged parturition and all signs of toxicity including mortality. Any changes were recorded including onset, degree and duration of signs as applicable.

DETAILED CLINICAL OBSERVATIONS:
More detailed examinations were made once before the first exposure (to allow for within-subject comparisons), then at least weekly, in the morning (am) or before treatment. These observations were made outside the home cage in a standard arena, at similar times as practical. The animals were monitored for changes in skin, fur, eyes, mucous membranes, occurrence of secretions and excretions, and autonomic activity (e.g. lachrymation, piloerection, pupil size, unusual respiratory pattern), or changes in gait, posture and response to handling as well as the presence of clonic or tonic movements, stereotypies (e.g. excessive grooming, repetitive circling), difficult or prolonged parturition or bizarre behaviour (e.g. self-mutilation, walking backwards); special attention were directed towards the observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma. No such clinical signs were observed during the study.

NEUROBEHAVIOURAL EXAMINATION
5 males and 5 females/group, “subgroup A”: Assessment of any potential test item related neurotoxicity was performed during the last exposure week (males on Day 24am; females on PPD 3am). In order to avoid hyperthermia of pups, dams were removed from the pups for not more than approximately 30-40 minutes. Selected animal were subjected to the functional observation battery, including qualitative assessment of the grip strength, and to measurements of the landing foot splay and fore/hind grip strength.

To measure the landing foot splay, the fore/hind paws of the rat were painted with ink and the rats were dropped from a horizontal position onto the appropriate record sheet covering the examination table. The distance between the two resulting ink spots was measured.

Fore/hind grip strength measurements were conducted using a grip strength meter (Model GS3, Bioseb, Chaville, France), an instrument designed to quantify objectively rodent muscular strength, in order to identify and assess quantitatively any potential effect of test item. The rats were held appropriately such that the fore limbs were allowed to grip the support bar and gently pulled back until they released the bar; the device measured the maximum grip strength. This was performed 3 times for each animal on each test day. The procedure was repeated with the hind limbs with the appropriate grip support; results are tabulated with individual and mean data.

Sensory reactivity to different type of stimuli (e.g. auditory, visual and proprioceptive), assessment of grip strength and motor activity were conducted and the general physical condition and behaviour of animals were tested. A modified Irwin test was performed. Parameters such as, but not limited to body position, locomotor activity, respiration rate, respiration type, piloerection, head searching compulsive biting or licking, circling, upright walking, retropulsion, jumping, exophthalmos, twitches, clonic convulsions, tonic convulsions, tremor, startle, transfer arousal, spatial locomotion, gait, posture, limb position, finger approach, finger withdrawal, touch escape response, diarrhoea, diuresis, visual placing, grip strength, body tone, corneal reflex, pinna, toe pinch, grasping reflex, positional struggle, skin, mucous membrane colour, salivation, palpebral closure, lachrymation, limb tone, abdominal tone, tail pinch, righting reflex, and/or vocalisation were evaluated.

BODY WEIGHT:
All adult animals were weighed with accuracy of 1 g for randomization purposes, then on Day 0, afterwards at least weekly and at termination. Parent females were weighed on gestation Days GD 0, 7, 14 and 20 and on postpartal Days PPD0 (within 24 hours after parturition), PPD4 and PPD5 (before termination). Body weights of the female animals were additionally weighed on gestational Days GD10 and 17 in order to give accurate treatment volumes, but these data were not evaluated statistically.

FOOD CONSUMPTION
Animal food consumption was determined by re-weighing the non-consumed diet with a precision of 1 g on Day 7 then at least weekly (see Study Schedule).

OPHTHALMOSCOPIC EXAMINATION:
The fundus of eyes of all animals was examined before treatment. Five male and 5 female Control and High dose animals (“subgroup A”) randomly selected from groups 1 and 4, during the last week of treatment prior to necropsy (males, Day 24pm, females, PPD 3 pm). Mydriasis was produced after instillation of a mydriatic agent (eye drops "Humapent") into the conjunctival sac. The examination was performed using a Gowlland ophthalmoscope. As no ophthalmoscopic alterations were found, no additional examination was performed in other animals.

OBSERVATION OF THE DELIVERY PROCESS, OFFSPRING AND NURSING INSTINCT
Females were allowed to litter and rear their offspring. Delivery process was observed as carefully as possible. All observations were recorded as applicable. No evidence of abnormal deliveries was recorded. The duration of gestation was recorded and was calculated from Day 0 of pregnancy. Dams were observed to record whether they form a nest from the bedding material and cover their new-borns or not. The efficiency of suckling was observed by the presence of milk in the pups' stomach. All observations were recorded as applicable.

Each litter was examined as soon as possible after delivery to establish the number and sex of pups, stillbirths, live births, runts (pups that are significantly smaller than normal pups) and the presence of gross abnormalities.

Observations are reported individually for each adult animal. In addition to the observations on parent animals, the pups (offspring) were monitored for any behavioural changes. Live pups were counted, sexed, weighed individually within 24 hours of parturition (ex. Day 0 or 1 post-natal, PND0 or 1) and on PND4, with accuracy of 0.01g. All the litters were checked and recorded daily for the number of viable and dead pups. The pups found dead and intact (not cannibalized) were subjected to necropsy with macroscopic examination in order to identify the possible cause of death. All observed abnormalities were recorded

HAEMATOLOGY:
All animals selected for blood sampling were fasted (overnight period of food deprivation). 5 males and 5 females/group, “subgroup B”: For terminal blood sampling of animals selected (subgroup B), 3 samples were taken from each scheduled animal: for haematology (in tubes with K3-EDTA as anticoagulant), one sample for blood clotting times (APTT and PT measurements, in tubes with sodium citrate as anticoagulant) and one to obtain serum (in tubes with no anticoagulant) for clinical chemistry. Blood samples were collected by cardiac puncture under pentobarbital anaesthesia, immediately prior to scheduled necropsy.-Parameters checked in table on page 22 of the report were examined.

CLINICAL CHEMISTRY:
- Time schedule for collection of blood: immediately prior to scheduled necropsy.
- Animals fasted: Yes
- How many animals: 5 males and 5 females
- Parameters checked in table on page 23 of the report were examined.

URINALYSIS:
Urine sampling (approximately 16 hours sampling period) was performed prior to necropsy (males on Day 28; females on PND 5). Parameters checked in table on page 24 of the report were examined.

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes

Fetal examinations:

FETAL EXAMINATIONS
Females were allowed to litter and rear their offspring. Delivery process was observed as carefully as possible. All observations were recorded as applicable. No evidence of abnormal deliveries was recorded. The duration of gestation was recorded and was calculated from Day 0 of pregnancy. Dams were observed to record whether they form a nest from the bedding material and cover their new-borns or not. The efficiency of suckling was observed by the presence of milk in the pups' stomach. All observations were recorded as applicable.

Each litter was examined as soon as possible after delivery to establish the number and sex of pups, stillbirths, live births, runts (pups that are significantly smaller than normal pups) and the presence of gross abnormalities.

GROSS EXAMINATION OF DEAD PUPS:
Observations are reported individually for each adult animal. In addition to the observations on parent animals, the pups (offspring) were monitored for any behavioural changes.

PARAMETERS EXAMINED
Live pups were counted, sexed, weighed individually within 24 hours of parturition (ex. Day 0 or 1 post-natal, PND0 or 1) and on PND4, with accuracy of 0.01g. All the litters were checked and recorded daily for the number of viable and dead pups. The pups found dead and intact (not cannibalized) were subjected to necropsy with macroscopic examination in order to identify the possible cause of death. All observed abnormalities were recorded.

Statistics:

Data were collected using the software PROVANTIS v.7, or were recorded on the appropriate forms from the relevant SOPs of CiToxLAB Hungary Ltd., then tabulated using the software PROVANTIS v.7, Microsoft Office Word and/or Excel, as appropriate. Numerical data obtained during the conduct of the study were subjected as appropriate to calculation of group means and standard deviations.

The statistical evaluation of appropriate data (marked † below) was performed with the statistical program package SPSS PC+4.0. The homogeneity of variance between groups was checked by Bartlett’s homogeneity of variance test. Where no significant heterogeneity was detected, a one-way analysis of variance (ANOVA) was carried out. If the obtained result was significant, Duncan Multiple Range test was used to access the significance of inter-group differences. Getting significant result at Bartlett’s test, the Kruskal-Wallis analysis of variance was used and the inter-group comparisons were performed using Mann-Whitney U-test. Chi2 test was performed as feasible.

Indices:

In parental males the following parameters were detemined: number of fertile pairings, number of infertile males, male mating index and male fertility index.

In parental females the following parameters were detemined: number of pairings, number of pregnant females, number of sperm positive, but non-pregnant females, number of non mated females, female mating index, female fertility index, gestation index, duration of pregnancy (days), number of corpora lutea/dams, number of implantations/dams, number of dams with live pups (day 0 and 4), pre-implantation mortality, intrauterine mortality,
total mortality (intra and extra uterine mortality).

In offspring the following parameters were detemined: mean pup body weight (per pup within the group and per litter) on PND 0 and 4, mean pup body weight gain (per litter) between postnatal Days 0-4, number of live births per litter, and number of viable pups per litter on postnatal Days 0 and 4, survival Index of pups on postnatal Days 0 and 4, sex ratio % (on postnatal Days 0 and 4).

Details on maternal toxic effects:

Maternal toxic effects:no effects

Dose descriptor:

NOAEL

Effect level:

> 1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

other: maternal toxicity

Dose descriptor:

NOAEL

Effect level:

> 1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

other: developmental toxicity

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Dose descriptor:

NOAEL

Effect level:

> 1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

other: fetotoxicity

Abnormalities:

not specified

Developmental effects observed:

not specified

Conclusions:

Under the conditions of this study, the no observed adverse effect level (NOAEL) for Epoxidized Soybean Oil Acrylate for parental/adult and F1 effects is considered to be 1000 mg/kg bw/day.

Executive summary:

The purpose of this Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test in the Rat study was to obtain information on the toxicity of the test item following repeated daily administration by oral gavage to Wistar rats. The study also included a reproductive/developmental toxicity-screening test, intended to provide initial information on possible effects on male and female reproductive performance such as gonadal function, mating behaviour, conception, pregnancy, parturition and also on the development of the F1 offspring from conception to Day 4 post-partum. The study was conducted according to OECD Guideline No. 422.

Male and female Wistar rats were treated for 2 weeks pre-mating, then during the mating/postmating period, males for 28 days and females throughout gestation period and up to and including postpartum/lactation Day PPD5

Daily administration of Epoxidized Soybean Oil Acrylate by oral gavage to Wistar rats did not result in test item related mortality or clinical adverse effects at daily, weekly or neurological assessment, in ophthalmological changes, or changes in the body weight, food consumption, haematology, coagulation, clinical chemistry, or urinalysis parameters at dose levels of 62.5, 250, or 1000 mg/kg bw/day during the treatment period under the conditions of this study.

 

No test item related, or adverse effects were noted at evaluation of the reproductive parameters during mating and gestation, delivery and post-partum/lactation period until PPD5, under the conditions of this study.There were no adverse effects ascribed to test item administration on the F1 offspring viability, clinical signs, development or at observations following euthanasia. There were no test item-related changes observed in organ weights, at necropsy or at histopathology for the adult animals of either sex.

 

In conclusion, under the conditions of this study, the no observed adverse effect level (NOAEL) for Epoxidized Soybean Oil Acrylate for parental/adult and F1 effects is considered to be 1000 mg/kg bw/day.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Additional information

The dossier does not currently have a pre-natal developmental toxicity study. Therefore, there is a data gap for this endpoint. However, the results of the OECD 422 reproduction/developmental toxicity screening study indicate no test item related, or adverse effects in any of reproductive parameters during mating and gestation, delivery and post-partum/lactation period. There were no adverse effects ascribed to test item administration on the F1 offspring viability, clinical signs, development or at observations following euthanasia. The NOAEL for parental/adult and F1 effects is considered to be 1000 mg/kg bw/day. Moreover, due to its high MW (>1000), it is very unlikely that the substance will be systemically absorbed and will cause any adverse effects in a full developmental toxicity study.

Justification for selection of Effect on developmental toxicity: via oral route:
There is only a screening reproduction/developmental toxicity study available. The study has been conducted according to OECD guideline no. 422 and under GLP.

Justification for classification or non-classification

No test item related, or adverse effects were noted at evaluation of the reproductive parameters during mating and gestation, delivery and post-partum/lactation period, under the conditions of this study.There were no adverse effects ascribed to test item administration on the F1 offspring viability, clinical signs, development or at observations following euthanasia. There were no test item-related changes observed in organ weights, at necropsy or at histopathology for the adult animals of either sex. The NOAEL for Epoxidized Soybean Oil Acrylate for parental/adult and F1 effects is considered to be 1000 mg/kg bw/day.

Therefore, there is no need to classify the substance for fertility and developmental toxicity, according to the Regulation EC 1272/2008 and the Directive 67/584/EEC.