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Toxicological information

Repeated dose toxicity: oral

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Administrative data

sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP guideline study with acceptable restriction. Purity not reported.

Data source

Reference Type:
study report
Report Date:

Materials and methods

Test guidelineopen allclose all
equivalent or similar to
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
Guideline not stated, purity not reported.
according to
other: Commission of the European Communities CS/PM/2024, 2 April 1993
GLP compliance:
Limit test:

Test material

Test material form:
other: solid, not further specified
Details on test material:
- Name of test material (as cited in study report): Stearyl erucamide
- Physical state: solid
- Analytical purity: not reported, responsibility of sponsor
- Lot/batch No.: 3751D
- Expiration date of the lot/batch: 18 May 1997
- Stability under test conditions: not examined
- Storage condition of test material: refrigerated at approximately 4 °C in the dark

Test animals

other: Crl: CD(SD)BR (VAF plus)
Details on test animals and environmental conditions:
- Source: Charles River (UK) Limited, Margate, England
- Age at study initiation: 5 to 6 weeks
- Weight at study initiation: males 141-176 g, females 112-148 g
- Fasting period before study: not applicable, feeding study
- Housing: groups of 5 by sex, in grid-bottomed stainless steel cages (Modular Systems, Kent, England), suspended over paper-lined trays
- Diet (e.g. ad libitum): Powdered diet (SQC Rat and Mouse Maintenance Diet No. 1, Ground Fine, Special Diets Services Limited, Witham, England), ad libitum (except for those periods of deprivation associated with laboratory investigations).
- Water (e.g. ad libitum): Mains water provided in polypropylene bottles, ad libitum (except for those periods of deprivation associated with laboratory investigations).
- Acclimation period: 12 days

- Temperature (°C): 18-25
- Humidity (%): 28-48
- Air changes (per hr): air conditioned, not further specified
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
oral: feed
unchanged (no vehicle)
Details on oral exposure:
- Rate of preparation of diet (frequency): Diet mixes were formulated weekly.
- Mixing appropriate amounts with (Type of food): Powdered diet (SQC Rat and Mouse Maintenance diet No. 1, Ground Fine, Special Diets Services, Witham, England). A pre-mix was prepared at an appropriate concentration by mixing a weighed amount of test article with a small quantity of diet. This was then mixed with more diet in a rotary mixer and made up to a final quantity with the remaining blank diet in a double-cone blender. To prepare formulated diets appropriate quantities of the pre-mix were mixed with blank diet to make up to the final quantity in a double-cone blender. Formulated diets for males and females were mixed separately.
- Storage temperature of food: not reported
Analytical verification of doses or concentrations:
Details on analytical verification of doses or concentrations:
To determine the achieved concentration, samples of each diet (including that for the control group), prepared for weeks 1 and 13, were analysed for test article by the Sponsor using a previously validated method. The results are reported separately by the Sponsor.
The stability and homogeneity of test article diets prepared over the concentration ranges used on this study have been assessed by the Sponsor, as well, using the previously validated method.

The achieved dose levels were within 3% of nominal. All individual group values were within 15% of nominal.
Duration of treatment / exposure:
13 weeks
Frequency of treatment:
continuously in diet
Doses / concentrations
Doses / Concentrations:
250, 500, 1000 mg/kg bw/day
nominal in diet
No. of animals per sex per dose:
Control animals:
yes, plain diet
Details on study design:
- Dose selection rationale: Dose levels were selected by the Sponsor after examining preliminary work performed by Quintiles (Quintiles Study Number: CRC/01/C).
Positive control:


Observations and examinations performed and frequency:
- Time schedule: Twice daily for mortality and morbidity, once daily for clinical signs; if required by clinical condition of an animal, monitoring was adjusted accordingly.
- Cage side observations included: Mortality, morbidity, clinical signs.

- Time schedule for examinations: On the day prior to dosing and then weekly thereafter.

- Food consumption for each group determined weekly, and mean daily diet consumption per animal calculated as g food/animal/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No

- Time schedule for examinations: Two days prior to dosing and on Day 85.
- Dose groups that were examined: All animals before dosing, and control and high dose group on Day 85

- Time schedule for collection of blood: Week 13
- Anaesthetic used for blood collection: No, blood was taken via tail vein puncture
- Animals fasted: Yes, overnight
- How many animals: All animals
- Parameters examined (in blood taken into EDTA anticoagulant): Erythrocyte count, hemoglobin concentration, leukocyte differential count, mean cell hemoglobin, mean cell hemoglobin concentration, mean cell volume, packed cell volume, platelet count, total leukocyte count.

- Time schedule for collection of blood: Week 13
- Animals fasted: Yes, overnight
- How many animals: All animals
- Parameters examined (in blood taken into lithium heparin anticoagulant): A/G ratio, alanine aminotransferase, albumin, alkaline phosphatase, aspartate aminotransferase, blood urea nitrogen, calcium, chloride, cholesterol, creatinine, glucose, phosphorus, potassium, sodium, total bilirubin, total protein. Coagulation tests (on blood taken into 3.2% aqueous trisodium citrate anticoagulant): activated partial thromboplastin time, fibrinogen, prothrombin time.

- Time schedule for collection of urine: Week 13
- Metabolism cages used for collection of urine: No data
- Animals fasted: Yes, overnight
- Parameters examined: Bilirubin, blood pigments, glucose, ketones, leukocytes, microscopic examination of sedimented deposit, nitrites, pH, protein, specific gravity, urobilinogen, volume.
Sacrifice and pathology:
GROSS PATHOLOGY: Yes; all animals were killed by CO2 asphyxiation, weighed and examined externally. The animals were then exsanguinated. The cranial, thoracic and abdominal cavities were opened and examined macroscopically. Any abnormalities were recorded with details of the location, colour, shape and size.

HISTOPATHOLOGY: Yes; with the exception of the eyes, optic nerves and testes, either whole organs or samples of the following tissues were preserved in neutral buffered formaldehyde. The eyes and optic nerves were fixed in Davidson's fluid and the testes were fixed in Bouin's fixative for 24 hours and then transferred to neutral buffered formaldehyde. Organs examined: adrenals, aorta, brain (3 sections), caecum, colon, duodenum, epididymides, eyes (incl. optic nerves), femur (incl. marrow) and joint, heart, ileum, jejunum, kidneys, liver, lungs (incl. mainstem bronchi), mesenteric lymph node, oesophagus, ovaries, pancreas, pituitary, prostate, rectum, salivary gland, sciatic nerve, seminal vesicles, site of mammary gland, skeletal muscle, skin, spinal cord (3 levels), spleen, sternum, stomach, submandibular lymph node, testes, thymus, thyroids, (incl. parathyroids), trachea, urinary bladder, uterus, vagina, all gross lesions. The aforementioned organs were examined from control and high dose animals and from all animals deceased or killed during the study; lungs and all gross lesions were examined from all animals. If target organs were identified in high dose animals they were examined in low and intermediate dose animals, as well.
Other examinations:
Absolute and relative organ weights from all animals: adrenals, brain, heart, kidneys, liver, ovaries, pituitary, spleen, testes, thymus, thyroids; contralateral organs were weighed together.
Group means and standard deviations were calculated where appropriate. No analysis will be performed for variable for which the control group has less than 5 independent observations per group. In situations where the treated groups have fewer than 5 observations comparisons will be performed at the discretion of the statistician. Sexes will be analysed separately. If combination of sexes is required for a particular analysis (for example because of small group sizes), then the analysis will be adjusted for sex differences as well as treatment group, and a test performed for a sex by treatment interaction. Such an analysis will be supported by scientific or statistical rationale. The data will be examined prior to analysis for any obvious outlying observations, which will be excluded from the analysis, any obvious skew in the distribution, for which the data will be log transformed prior to analysis and also for a high incidence of tied data. If the proportion of ties in a data set exceeds 30% then the non-parametric methods outlined below will be deemed unsuitable. Any adhoc comparisons performed, other than those described below, will be performed using a suitable conservative comparison procedure.

Results and discussion

Results of examinations

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
unrelated to treatment
mortality observed, treatment-related
Description (incidence):
unrelated to treatment
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
unrelated to treatment
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
no effects observed
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
not toxicologically significant
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
not toxicologically significant
Urinalysis findings:
effects observed, treatment-related
Description (incidence and severity):
unrelated to treatment
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
not toxicologically significant
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
unrelated to treatment
Histopathological findings: neoplastic:
no effects observed
Details on results:
Hairloss was noted in the treated animals, which is a common finding in animals of this strain and age and lacked any dose-relationship.

One mid dose female was killed in extremis on day 50, clinical signs included peribuccal swelling, teeth abnormalities and black periorbital staining. Histopathology revealed peribuccal ulceration. In view of the isolated nature of this finding it was considered to be unrelated to treatment.

Bodyweight and bodyweight gains were considered to be unaffected by treatment. Body weight gains were slightly reduced in all treated male groups, however these values were within 8% of controls and therefore considered not to be an effect of treatment. Female groups showed bodyweight gains within 3% of controls. The slight bodyweight loss noted in week 13 of the study was considered to be attributable to blood sampling procedures.

Food consumption was unaffected by treatment and was similar in all groups.

There were no treatment-related findings at ophthalmoscopic examination.

A non-dose-related increase in monocytes was apparent in all treated male groups. Group mean values for the low and the mid dose groups were within the expected background ranges. However, 5/10 males of the high dose group exceeded the normal range. Due to the small magnitude of this change and in the absence of any related pathological changes, this was considered to be without toxicological significance.

All treated females showed reduced activated partial thromboplastin times compared to controls; however, this was considered to be due to two animals in the control group having high values and not to any effect of the test substance.

There were no changes considered to be of toxicological significance. The statistically significant values noted in several parameters (creatinine, albumin) were minor, and all group mean values were within the normal ranges found in this laboratory.

There were no changes attributable to treatment. The trace and small amounts of epithelial cells and casts noted were apparent throughout the dose groups, and in the absence of any supporting pathology this was considered to be unrelated to the test substance.

A statistically significant reduction in absolute liver weights was observed in all treated male groups, and a statistically significant reduction in bodyweight-related liver weights was observed in males of the mid and high dose group. In the absence of corresponding histopathological findings this was considered not to be of toxicological significance.

A statistically significant reduction in absolute brain weight was apparent in all treated males. However, there was no effect on bodyweight-related brain weights, and therefore this finding was considered to be unrelated to treatment.

Body weight-related heart weights were statistically higher than in controls in all treated female groups. As this change was minor and lacked a dose-related manner, it was considered not to be of any toxicological significance.

There were no treatment-related macroscopic findings.

There were no treatment-related microscopic findings. The small number of histopathological findings recorded were within the normal range of background alterations which may be seen in untreated rats of this age and strain.

There were no neoplastic findings reported.

Effect levels

Dose descriptor:
Effect level:
>= 1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
other: No adverse effects observed up to the highest dose tested.

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables


The repeated daily administration of the test substancein the diet to the Crl:CD(SD)BR(VAF plus) strain for 13 weeks at dose levels of 250, 500 and 1000 mg/kg bw/day did not reveal any evidence of toxicity. Therefore, the NOAEL was determined to be 1000 mg/kg bw/day.

Applicant's summary and conclusion