Soybean oil, epoxidized, acrylate

Endpoint summary

Description of key information

No effects were observed with the test substance up to 1000 mg/kg bw (the highest dose level used) in a repeated dose toxicity/screening reprotoxicity study according to OECD guideline No. 422. 

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: oral

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:

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:

no

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 oral 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.

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.

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).

Remarks:

Doses / Concentrations:
62.5, 250 and 1000 mg/kg bw
Basis:
other: nominal concentrations based on test material

No. of animals per sex per dose:

12 animals/sex/dose

Control animals:

yes, concurrent vehicle

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.

Positive control:

not applicable

Observations and examinations performed and frequency:

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.

Sacrifice and pathology:

GROSS PATHOLOGY:
Surviving animals were euthanized upon completion of the study, according to the study plan.

Gross necropsy was performed on all animals. Terminally, after completion of the treatment, animals were euthanised under pentobarbital anaesthesia, followed by exsanguination (see "Details of Other Materials"). 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. Special attention was paid to the organs of the reproductive system. The number of implantation sites and of corpora lutea was recorded in the females as applicable.

At the time of termination, body weight and weight of the following organs of all parental animals were determined:
- With a precision of 0.01 g: uterus (with and without cervix), vagina, testes, epididymides (total and cauda), prostate, seminal vesicles with coagulating glands, brain
- With a precision of 0.001 g: ovaries, pituitary

The weighed organs and all organs showing macroscopic lesions of all adult animals were preserved. The eyes with the optic nerve were retained in modified Davidson’s fixative. Testes and epididymides were preserved in Bouin’s solution, all other organs in 10% buffered formalin solution.

In addition, for 5 animals/sex/group (subgroup B), the following organs and tissues, or representative samples, were preserved:
Gross findings, Adrenals, Animal identification, Aorta, Brain, Epididymides, Eyes with optic nerves, Oesophagus, Femur with marrow , Heart, Kidneys, Large intestine, Lacrimal glands, Harderian glands, Liver, Lungs with bronchi, Lymph nodes, Mammary gland (inguinal), Ovaries with oviduct, Pancreas, Pituitary, Prostate, Salivary gland (mandibular), Sciatic nerve, Seminal vesicles, Coagulating glands, Skeletal muscle (quadriceps), Skin and subcutis (inguinal),
Small intestine, Spinal cord (cervical, lumbar, and thoracic levels), Spleen, Sternum with marrow, Stomach, Testes, Thymus, Thyroid with parathyroids,
Tongue, Trachea (with main stem bronchi), Urinary bladder, Uterus, Vagina.

From subgroup B animals, the following organs were weighed in addition to the ones previously mentioned:
- With a precision of 0.01 g: heart, kidneys, liver, spleen and thymus.
- With a precision of 0.001 g: adrenals.

For all organs, paired organs were weighed individually. Individual and/or paired absolute organ weight are reported for each animal and adjusted for the body and brain weights. Paired organ weights as applicable were summarised. Relative organ weight (to body and brain weight) were calculated and reported.

HISTOPATHOLOGY:
For the adult animals, detailed histological examination was performed on the selected list of retained organs in the Control and High dose groups and all macroscopic findings (abnormalities) from all animals. The retained tissues and organs were embedded in paraffin wax, sections were cut at 4-6µ by microtome and transferred to slides. Tissue sections were stained with haematoxylin-eosin/phloxine and examined by light microscope.

As no test item related pathology findings were noted, no additional histopathology evaluation was considered required. Special attention was paid to evaluation of the stages of spermatogenesis in the male gonads and histopathology of interstitial testicular cell structure. Detailed histological examination of the ovaries covered the follicular, luteal, and interstitial compartments of the ovary, as well as the epithelial capsule and ovarian stroma.

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.

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

no effects observed

Water consumption and compound intake (if drinking water study):

no effects observed

Ophthalmological findings:

no effects observed

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

no effects observed

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

not specified

Details on results:

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.

GROSS PATHOLOGY
At necropsy, no treatment-related macroscopic findings were observed. Changes such as small testes and epididymides (in one control male), or firm mass at the mammary gland region, enlarged spleen and enlarged/cystic ovaries (in one high dose female) were incidental.

HISTOPATHOLOGY
No treatment-related microscopic findings were observed. The minimal, multifocal congestion/haemorrhage in the thymus, the minimal, focal/multifocal, perivascular, mononuclear cell infiltrate of the prostate, the tubular degeneration/atrophy in the testes, or the diffuse aspermia in the epididymides, could be observed across control and treated groups, and were therefore regarded as incidental or common background, not toxicologically significant and/or not associated with the test item administration.

The galactocele characterized as dilatation of ducts and acini, contained proteinaceous, eosinophilic material, without evidence of epithelial proliferation was seen in the mammary gland of one high dose female . There were no other microscopic changes in reproductive organs including the ovaries, uterus and vagina in this female. The galactocele was considered as spontaneous change. In addition, extramedullary hematopoiesis in the spleen (in correlation with necropsy) was also recorded for this female rat.

Dose descriptor:

NOAEL

Remarks:

for parental/adult toxicity

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Critical 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.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

The dossier does not currently have a subchronic (90-day) toxicity study. Therefore, there is a data gap for this endpoint. However, the results of the OECD 422 combined repeated dose/reproduction and developmental toxicity screening study indicate no test item related or adverse effects. Daily administration of the test substance 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. 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 parental/adult and F1 effects is considered to be 1000 mg/kg bw/day in this study. It is, therefore, very unlikely that the substance will cause any adverse effects in a subchronic (90-day) study.

Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
This is the only available repeated dose toxicity. The study has been conducted according to OECD guideline 422 and under GLP.

Justification for classification or non-classification

No effects were observed up to 1000 mg/kg bw (the highest dose level used) in a repeated dose toxicity/screening reprotoxicity study according to OECD guideline No. 422. Therefore, the substance does not need to be classified for repeated dose toxicity, according to the Regulation EC 1272/2008 and the Directive 67/584/EEC.