Registration Dossier

Administrative data

Endpoint:
one-generation reproductive toxicity
Remarks:
based on generations indicated in Effect levels (migrated information)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
09 March - 23 April 2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was performed according to GLP and OECD guidelines.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report date:
2010

Materials and methods

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

Test material

Constituent 1
Reference substance name:
Rape oil, oxidized
EC Number:
305-871-3
EC Name:
Rape oil, oxidized
Cas Number:
95193-59-2
IUPAC Name:
95193-59-2
Details on test material:
- Name of test material (as cited in study report): Blown rapeseed oil
- Substance type: Yellow to brown viscous liquid
- Physical state: liquid
- Analytical purity: treated as 100% pure
- Expiration date of the lot/batch: 16 December 2010
- Stability under test conditions: stable
- Storage condition of test material: At room temperature in the dark
-Other: Specific Gravity 0.9660 g/cm3 (20°C)
-Other: pH (1% in water, indicative range) 4.9-4.7 (determined at NOTOX)

Test animals

Species:
rat
Strain:
other: Crl:WI(Han)
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: F0 Charles River Laboratories France, L'Arbresle Cedex. France.
- Age at study initiation: Approximately 11 weeks
- Weight at study initiation: no data
- Fasting period before study: no
- Housing:
Pre-mating: Animals were housed in groups of 5 animals/sex/cage in Macrolon plastic cages (MIV type, height 18 cm).
Mating: Females were caged together with males on a one-to-one-basis in Macrolon plastic cages (MIII type, height 18 cm).
Post-mating: Males were housed in their home cage (Macrolon plastic cages, MIV type, height 18 cm) with a maximum of 5 animals/cage. Females were individually housed in Macrolon plastic cages (MIII type, height 18 cm).
Lactation: Pups were kept with the dam until termination in Macrolon plastic cages (MIII type, height 18 cm).
General: Sterilised sawdust as bedding material (Litalabo, S.P.P.S., Argenteuil, France) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom) were supplied. Certificates of analysis were examined and then retained in the NOTOX archives. During activity monitoring, animals were housed individually in Macrolon plastic cages (MIII type; height 15 cm) with sterilized sawdust as bedding material. No cage-enrichment was provided during activity monitoring.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: At least 5 days prior to start of treatment

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.2 – 21.4°C
- Humidity (%): 23 - 89%
- Air changes (per hr): approximately 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours artificial light and 12 hours darkness per day.
Cleaning procedures in the room might have caused the temporary fluctuations above the optimal maximum level of 70% for relative humidity (with a maximum of 4 hours). Temporary fluctuations from the light/dark cycle (with a maximum of 1 hour) occurred due to performance of pupillary reflex tests in the room. Based on laboratory historical data, these fluctuations were considered not to have affected the study integrity.

IN-LIFE DATES: From: 09 March 2010 To: 23 April 2010



Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
Formulations (w/w) were prepared daily within 6 hours prior to dosing and were homogenised to a visually acceptable level. Adjustment was made for specific gravity of the test substance and vehicle.

VEHICLE
- Justification for use and choice of vehicle (if other than water): Based on trial formulations performed at NOTOX.
- Concentration in vehicle: 75, 225 and 500 mg/ml
- Amount of vehicle (if gavage): 2 mL/kg body weight
Details on mating procedure:
- M/F ratio per cage: 1/1
- Length of cohabitation: A maximum of 14 days was allowed for mating. After 14 days of mating, female 46 (Group 1) had not shown evidence of mating and was separated from her male. However, mating of this female was inadvertently overlooked, since she later delivered live offspring.
- Proof of pregnancy: confirmed by evidence of sperm in the vaginal lavage, by staging of the estrous cycle and/or or by the appearance of an intravaginal copulatory plug. This day was designated Day 0 post-coitum. Once mating occurred, the males and females were separated.
- After successful mating each pregnant female was caged individually
Analytical verification of doses or concentrations:
no
Details on analytical verification of doses or concentrations:
No analyses were conducted since no analytical method could be developed for formulations of the test substance in corn oil.
Duration of treatment / exposure:
Males were exposed for 29 days, i.e. 2 weeks prior to mating, during mating, and up to termination. Females were exposed for 41-45 days, i.e. during 2 weeks prior to mating, during mating, during post-coitum, and during at least 4 days of lactation. Females 69 (Group 3) and 73 (Group 4) were not dosed during littering.
Frequency of treatment:
Once daily for 7 days per week, approximately the same time each day with a maximum of 6 hours difference between the earliest and latest dose. Animals were dosed up to the day prior to scheduled necropsy.
Details on study schedule:
- Age at mating of the mated animals in the study: Approximately 13-14 weeks
Doses / concentrations
Remarks:
Doses / Concentrations:
0, 150, 450 and 1000 mg/kg/day
Basis:
actual ingested
No. of animals per sex per dose:
10 animals/sex/dose + 460 pups in total
Details on study design:
- Dose selection rationale: Dose levels were based on results of a 10-Day dose range finding study (NOTOX Project 493033)
-Results of the dose range finding study are described in End point study record Repeated dose toxicity: oral.Notox Project 493025.
-5 animals/sex/group were randomly selected at allocation for functional observations, clinical pathology, macroscopic examination (full list), organ weights (full list) and histopathology:
Males: the first 5 males per group
Females: with live offspring
- Post-exposure recovery period in satellite groups: none
Positive control:
none

Examinations

Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: At least twice daily (early morning/late afternoon)

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Detailed clinical observations were made in all animals, immediately after dosing. Once prior to start of treatment and at weekly intervals this was also performed outside the home cage in a standard arena. Arena observations were not performed when the animals were mating, or housed individually. On March 30th 2010, no arena observations were performed for the males, but sufficient data was available from other observations.

BODY WEIGHT: Yes
- Time schedule for examinations: Males and females were weighed on the first day of exposure and weekly thereafter. Mated females were weighed on Days 0, 4, 7, 11, 14, 17 and 20 post-coitum, and during lactation on Days 1 and 4. No body weight was determined during the post-coitum period for female no. 46 (Group 1) because mating of this female was overlooked. Sufficient data was available for an accurate evaluation.

FOOD CONSUMPTION: Yes
-Weekly, except for males and females which were housed together for mating. Food consumption of mated females was measured on Days 0, 4, 7, 11, 14, 17 and 20 post-coitum and on Days 1 and 4 of lactation. No food consumption was determined during the post-coitum period for female no. 46 (Group 1) because mating of this female was overlooked. Sufficient data was available for an accurate evaluation.

WATER CONSUMPTION: No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: immediately prior to scheduled post mortem examination, between 7.00 and 10.30 a.m.
- Anaesthetic used for blood collection: Yes (iso-flurane)
- Animals fasted: Yes, but water was provided
- How many animals: 5 animals/sex/group (females used had live offspring)
- Parameters checked were: White blood cells, Differential leucocyte count (neutrophils, lymphocytes, monocytes,
eosinophils, basophils), Red blood cells, Reticulocytes, Red blood cell distribution width, Haemoglobin, Haematocrit, Mean corpuscular volume, Mean corpuscular haemoglobin, Mean corpuscular haemoglobin concentration, Platelets, Clotting Potential, Prothrombin time, Activated Partial thromboplastin time
Because of a possible switch of blood samples from female no. 45 (Group 1) and female no. 54 (Group 2) the rults for haematology and clinical biochemistry were excluded. These data were excluded and sufficient data from other animals is available to make an accurate evaluation.


CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: immediately prior to scheduled post mortem examination, between 7.00 and 10.30 a.m.
- Animals fasted: Yes, but water was provided
- How many animals: 5 animals/sex/group
- Parameters checked were: Alanine aminotransferase, Aspartate aminotransferase, Alkaline phosphatase, Total Protein, Albumin, Total Bilirubin, Urea
Creatinine, Glucose, Cholesterol, Sodium, Potassium, Chloride, Calcium, Inorganic Phosphate, Bile acids
Because of a possible switch of blood samples from female no. 45 (Group 1) and female no. 54 (Group 2) the rults for haematology and clinical biochemistry were excluded. These data were excluded and sufficient data from other animals is available to make an accurate evaluation.

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: The selected males were tested during Week 4 of treatment and the selected females were tested during lactation (all before blood sampling).
- Dose groups that were examined: all
- Battery of functions tested: hearing ability, pupillary reflex, static righting reflex, grip strength, motor activity:
Oestrous cyclicity (parental animals):
Not determined
Sperm parameters (parental animals):
Parameters examined in all male parental animals:
testis weight, epididymis weight.
For 5 males of the control and high dose group, slides of the testes were prepared to examine the staging of spermatogenesis

OTHER:
Male number paired with, mating date, confirmation of pregnancy, and delivery day was recorded. Pregnant females were examined to detect signs of difficult or prolonged parturition, and cage debris of these females was examined to detect signs of abortion or premature birth. Any deficiencies in maternal care (such as inadequate construction or cleaning of the nest, pups left scattered and cold, physical abuse of pups or apparently inadequate lactation or feeding) were examined.
Litter observations:
PARAMETERS EXAMINED
Each litter was examined to determine the following, if practically possible:

Mortality / Viability: The numbers of live and dead pups on Day 1 of lactation and daily thereafter were determined. If possible, defects or cause of death were evaluated.

Clinical signs: At least once daily, detailed clinical observations were made in all animals.

Body weights: Live pups were weighed on Days 1 and 4 of lactation.
There were no bodyweights and clinical observations performed for pup 15 from litter 43 (Group 1) on Day 1 because this pup was not seen on Day 1 because this pup was not seen on Day 1 or was born after the first litter check was conducted. This pup was observed on Day 2 onwards and was weighed on Day 4, and sufficient data is available for an accurate evaluation.

Sex: Sex was determined for all pups on Days 1 and 4 of lactation.

GROSS EXAMINATION OF DEAD PUPS: Yes
All pups were sexed and descriptions of all external abnormalities were recorded. The stomach was examined for the presence of milk. If possible, defects or cause of death were evalutated.
Postmortem examinations (parental animals):
GROSS PATHOLOGY: Yes
All animals were subjected to macroscopic examination of the cranial, thoracic and abdominal tissues and organs, with special attention being paid to the reproductive organs. Descriptions of all macroscopic abnormalities were recorded. The numbers of former implantation sites and corpora lutea were recorded for all paired females.
Samples of the following tissues and organs were collected and fixed in 10% buffered formalin (neutral phosphate buffered 4% formaldehyde solution, Klinipath, Duiven, The Netherlands):

From the selected 5 animals/sex/group:
Identification marks (not processed), Adrenal glands, All gross lesions, (Aorta), Brain (cerebellum, mid-brain, cortex), Caecum, Cervix, Clitoral gland, Colon, Coagulation gland, Duodenum, Epididymides (2), (Esophagus), Eyes (with optic nerve (if detectable) and Harderian gland; 2), Female mammary gland area, Femur including joint, Heart, Ileum, Jejunum, Kidneys, (Larynx), (Lacrimal gland, exorbital), Liver, Lung, infused with formalin, Lymph nodes (mandibular, mesenteric), (Nasopharynx), Ovaries, (Pancreas), Peyer's patches (jejunum, ileum) if detectable, Pituitary gland, Preputial gland, Prostate gland, Rectum, (Salivary glands - mandibular, sublingual), Sciatic nerve, Seminal vesicles, Skeletal muscle, (Skin), Spinal cord (cervical, midthoracic, lumbar), Spleen, Sternum with bone marrow, Stomach, Testes (2), Thymus, Thyroid including parathyroid (if detectable), (Tongue), Trachea, Urinary bladder, Uterus, Vagina

All remaining animals and females which failed to deliver**:
All gross lesions, Cervix, Clitoral gland, Coagulation gland, Epididymides*, Identification marks (not processed), Ovaries, Preputial gland, Prostate gland, Seminal vesicles, Testes*, Uterus, Vagina

* Fixed in modified Davidson's solution (prepared at NOTOX using Formaldehyde 37-40%, Ethanol, Acetic acid (glacial) (all Merck, Darmstadt, Germany) and Milli-Ro water (Millipore Corporation, Bedford, USA)) and transferred to formalin after fixation for at least 24 hours.
** In case no macroscopically visible implantation sites were present, nongravid uteri were stained using the Salewski technique (Ref. 1) in order to detect any former implantation sites (Salewski staining prepared at NOTOX using Ammoniumsulfide-solution 20% (Merck, Darmstadt, Germany) and Milli-Ro water (Millipore Corporation, Bedford, USA)).

Tissues/organs mentioned in parentheses were not examined by the pathologist, since no signs of toxicity were noted at macroscopic examination.

Four females (nos. 59, 60, 68, 78) were fasted approximately 3 hours longer than the maximum of 20 hours allotted before necropsy. The fasting period was only slightly longer and was considered not to have adversely affected the study's integrity.


HISTOPATHOLOGY: Yes
The delegated phase was performed by the Principal Investigator for histopathology.

The following slides were examined by a pathologist:
- The preserved organs and tissues of the selected 5 animals/sex of Groups 1 and 4.
- The additional slides of the testes of the selected 5 males of Groups 1 and 4 to examine staging of spermatogenesis.
- All gross lesions of all animals (all dose groups).
- The reproductive organs* of all animals that failed to mate, conceive, sire or deliver healthy pups (female no. 57 and male no. 17, Group 2).
* Reproductive organs included the cervix, clitoral gland, coagulation gland, epididymides, ovaries, preputial gland, prostate gland, seminal vesicles, testes, uterus, and vagina.

All abnormalities were described and included in the report. An attempt was made to correlate gross observations with microscopic findings.
Postmortem examinations (offspring):
SACRIFICE
Pups surviving to the planned termination were killed by decapitation on Days 5-6.

GROSS NECROPSY
All pups were sexed and descriptions of all external abnormalities were recorded. The stomach was examined for the presence of milk. If possible, defects or cause of death were evaluated. Any abnormal pup, organ or tissue was preserved in 10% buffered formalin for possible further examination.

HISTOPATHOLOGY / ORGAN WEIGHTS
None
Statistics:
The following statistical methods were used to analyze the data:
- If the variables could be assumed to follow a normal distribution, the Dunnett-test (many-to-one t-test, Dunnett 1955) based on a pooled variance estimate was applied for the comparison of the treated groups and the control groups for each sex.
- The Steel-test (many-to-one rank test; Miller 1981) was applied if the data could not be assumed to follow a normal distribution.
- The Fisher Exact-test (Fisher, 1950) was applied to frequency data.

The number of implantation sites was subjected to the Kruskal-Wallis nonparametric ANOVA test (Kruskal and Wallis, 1952) to determine intergroup differences followed by the Wilcoxon test (Wilcoxon, 1945) to compare the treated groups to the control group.

All tests were two-sided and in all cases p < 0.05 was accepted as the lowest level of significance.

Group means were calculated for continuous data and medians were calculated for discrete data (scores) in the summary tables. Test statistics were calculated on the basis of exact values for means and pooled variances. Individual values, means and standard deviations may have been rounded off before printing. Therefore, two groups may display the same printed means for a given parameter, yet display different test statistics values.
Reproductive indices:
For each group, the following calculations were performed:

Percentage Mated females: Number of females mated/Number of females paired x 100
Fertility index: Number of pregnant females/Number of females paired x 100
Conception index (%): Number of pregnant females/Number of females mated x 100
Gestation index (%): Number of females bearing live pups/Number of pregnant females x 100
Duration of gestation: Number of days between confirmation of mating and the beginning of parturition
Offspring viability indices:
Percentage live males at First Litter Check: Number of live male pups at First Litter Check/Number of live pups at First Litter Check x 100

Percentage live females at First Litter Check: Number of live female pups at First Litter Check/Number of live pups at First Litter Check x 100

Percentage of postnatal loss Days 0-4 of lactation: Number of dead pups on Day 4 of lactation/Number of live pups at First Litter Check x 100

Viability index (%): Number of live pups on Day 4 of lactation/Number of pups born alive x 100

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

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:
not examined

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
not examined
Reproductive function: sperm measures:
no effects observed
Reproductive performance:
no effects observed

Details on results (P0)

MORTALITY AND CLINICAL SIGNS (PARENTAL ANIMALS)
No mortality occurred during the study period.
No clinical signs of toxicity were noted during the observation period.

Incidental findings that were noted included scabbing of the left cheek and alopecia. These findings occurred within the range of background findings to be expected for rats of this age and strain which are housed and treated under the conditions in this study. At the incidence observed, these were considered signs of no toxicological relevance.

BODY WEIGHT AND WEIGHT GAIN (PARENTAL ANIMALS)
Body weights and body weight gain of treated animals remained in the same range as controls over the treatment period.

FOOD CONSUMPTION AND FOOD EFFICIENCY (PARENTAL ANIMALS)
Food consumption before or after allowance for body weight was similar between treated and control animals.

The statistically significant increase in relative food consumption seen for males of at 450 mg/kg (Group 3) was not considered to be toxicologically relevant. The higher relative food consumption occurred in the absence of a dose-related distribution, and if treatment related toxicity were evident, a reduction in food consumption would be expected to be seen.

Very low food consumption was seen for female nos. 45 (Group 1) and 61 (Group 3) over Days 0-4 of the post coitum period, however, the low weights were transient and body weight gain for both of these animals was normal over the same period, thus the low values were not considered to be adverse.

HAEMATOLOGY (PARENTAL ANIMALS)
There were no differences noted in haematological parameters between control and treated rats that were considered to be related to treatment with Blown rapeseed oil.

The statistically significant decrease in the activated partial thromboplastin time (APTT; males) at 1000 mg/kg compared to controls was not considered to be toxicologically relevant as the values for concurrent controls were slightly high and values at 1000 mg/kg remained within the range considered normal for rats of this age and strain.

The increase in neutrophil counts with concurrently reduced lymphocyte counts seen for animal no. 32 (Group 4, 1000 mg/kg) were not considered to be toxicologically relevant as it occurred only for an individual animal. This shift in type of white blood cells was considered to be a secondary non-specific response to stress and not to be treatment related.

CLINICAL CHEMISTRY (PARENTAL ANIMALS)
No toxicologically relevant changes occurred in clinical biochemistry parameters of treated rats.

The significantly higher inorganic phosphate level seen for females at 150 mg/kg was considered to be of no toxicological significance as it occurred in the absence of a treatment-related distribution and remained within the range considered normal for rats of this age and strain.

NEUROBEHAVIOUR (PARENTAL ANIMALS)
Hearing ability, pupillary reflex, static righting reflex and grip strength were normal in all animals tested.

The variation in motor activity did not indicate a relation with treatment. Despite not reaching statistical significance, the mean high sensor count for females is increased at 1000 mg/kg. This is attributable to the high counts for female no. 74. The very high values for this female may be due to extraneous factors like movement of the tail or head bobbing. If you discount this female’s data for high counts, the mean for Group 4 is 156, and thus, comparable to control values.

ORGAN WEIGHTS (PARENTAL ANIMALS)
No toxicologically relevant changes were noted in organ weights and organ to body weight ratios.

The significantly kidney to body weight ratios seen for both sexes at 450 mg/kg compared to control animals were considered not to be a sign of toxicity because they remained within the range considered normal for animals of this age and strain and occurred in the absence of a dose-dependent distribution and any microscopic indications of toxicity.

Other organ weights and organ to body weight ratios among the dose groups were similar to control levels.

GROSS PATHOLOGY (PARENTAL ANIMALS)
Macroscopic observations at necropsy did not reveal any alterations that were considered to have arisen as a result of treatment.

Incidental findings among control and treated animals included a focus/foci on the lungs, stomach glandular mucosa, and/or preputial glands, reddish or dark red discoloration of the stomach glandular mucosa, mesenteric and mandibular lymph nodes and/or thymus, discoloration of the preputial glands, enlargement of the mandibular lymph node, soft or hard nodule/nodules on the tail of the epididymis, clitoral glands, and/or left uterine horn, uterus contains fluid, diaphragmatic hernia of the liver, and alopecia. The incidence of these findings was within the background range of findings that are encountered among rats of this age and strain, and did not show a dose-related incidence trend. These necropsy findings were therefore considered to be of no toxicological relevance.

HISTOPATHOLOGY (PARENTAL ANIMALS)
There were no treatment-related microscopic findings.

No abnormalities were seen in the reproductive organs of the suspected non-fertile animals (Group 2 female 57 and male 17) which could account for their infertility.

All microscopic findings recorded were considered to be within the normal range of background pathology encountered in Wistar-Han rats of this age and strain.

REPRODUCTIVE FUNCTION: SPERM MEASURES (PARENTAL ANIMALS)
Staging of spermatogenesis did not provide any evidence of impairment of the spermatogenetic cycle.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
No toxicologically relevant effects on reproductive parameters were noted.
The mating, fertility and conception indices, precoital time, and number of corpora lutea and implantation sites were unaffected by treatment.

No toxicologically relevant effects on gestation index and duration, parturition, maternal care and early postnatal pup development (mortality, clinical signs, body weight and macroscopy) were observed.

Gestation
The gestation index and duration of gestation remained in the same range between controls and treated animals.

Parturition/Maternal care
No signs of difficult or prolonged parturition were noted among the pregnant females.
Examination of cage debris of pregnant females revealed no signs of abortion or premature birth and no deficiencies in maternal care were observed.

Effect levels (P0)

Dose descriptor:
NOAEL
Effect level:
> 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female

Results: F1 generation

General toxicity (F1)

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:
no effects observed

Details on results (F1)

VIABILITY (OFFSPRING)
Mortality
One pup of the control group, one pup at 450 mg/kg and one pup at 1000 mg/kg/day were found dead or missing on Day 2 of the lactation period. The control pup that was missing on Day 2 was most likely cannibalized. No toxicological relevance was attributed to these dead/missing pups since the mortality incidence did not show a dose-related trend and remained within the range considered normal for pups of this age.

CLINICAL SIGNS (OFFSPRING)
Incidental clinical symptoms of pups consisted of small or very small size, a blue-red spot on the head, a wound and scabbing on the abdomen and no milk in the stomach. The nature and incidence of these clinical signs remained within the range considered normal for pups of this age, and were therefore considered to be of no toxicological relevance.

BODY WEIGHT (OFFSPRING)
Body weights of pups were considered to have been unaffected by treatment.

GROSS PATHOLOGY (OFFSPRING)
There were no macroscopic findings for the individual pups at 450 and 1000 mg/kg that were found dead. Incidental macroscopic findings among surviving pups included small or very small size, a blue-red spot on the head and a scab on the abdomen. The nature and incidence of these findings remained within the range considered normal for pups of this age, and were therefore considered to be of no toxicological relevance.


Effect levels (F1)

Dose descriptor:
NOAEL
Generation:
F1
Effect level:
> 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female

Overall reproductive toxicity

Reproductive effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
Blown rapeseed oil was administered by daily oral gavage to male and female Wistar Han rats at dose levels of 150, 450 and 1000 mg/kg/day. Males were exposed for 2 weeks prior to mating, during mating, and up to termination (for 29 days). The females were exposed for 2 weeks prior to mating, during mating, during post-coitum, and at least 4 days of lactation (for 41-45 days).

Parental results:
No parental toxicity was observed at any dose level.

No treatment-related changes were noted in any of the parental parameters investigated in this study (i.e. clinical appearance, functional observations, body weight, food consumption, clinical laboratory investigations, macroscopic examination, organ weights, and microscopic examination).

Reproductive/Developmental results:
No reproduction or developmental toxicity was observed at any dose level.


In conclusion, treatment with blown rapeseed oil by oral gavage in male and female Wistar Han rats at dose levels of 150, 450 and 1000 mg/kg body weight/day revealed no parental toxicity up to 1000 mg/kg body weight/day. No reproduction and developmental toxicity was observed for treatment up to 1000 mg/kg body weight/day.

Based on these results, a parental, reproduction and developmental No Observed Adverse Effect Level (NOAEL) of at least 1000 mg/kg/day was derived.