Registration Dossier

Administrative data

Description of key information

Oral route: The NOAEL from a 90 day, repeated dose toxicity (oral route) of the read across substance is greater than or equal to 1,000 mg/kg/d.
Inhalation and Dermal route: A short term toxicity study does not need to be conducted because a reliable sub-chronic (90 days) or chronic toxicity study is available, conducted with an appropriate species,dosage, solvent and route of administration. This endpoint is waived.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
sub-chronic toxicity: oral
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
From 6th of August to 5th of November, 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Reason / purpose for cross-reference:
read-across: supporting information
Qualifier:
according to guideline
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)
GLP compliance:
yes (incl. QA statement)
Limit test:
no
Species:
rat
Strain:
other: Wistar, derived SPF- Albino, Crt: Wi/Br
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Versuchstierzucht, Charles River Wiga, 8741 Sulzfeld
- Age at study initiation: about 5 weeks (born the 1st and 2st July, 1987)
- Weight at study initiation: Males from 129 g to 170 g; Females from 115 g to 138 g
- Date of receipt: July 30th, 1987
- Animal health: at the entrance, at the test initiation. Certification available.
- Housing: in makrolon R cages.
- Bedding: "Altromin Laboreinstreu" of pure soft wood, dried, disdusted and sterilized at 180°C, supplied by Altromin GmbH, 4937 Lage, renewed weekly.
- Diet: Ssniff R pelletted diet (standard laboratory rat diet), ad libitum
- Diet quality assurance: diet analysed periodically from chlorinated hydrocarbons, aflatoxins, heavy metals, arsenic, and antibiotic activity (by "Landwirtschaf tliche Untersuchungs' und Forschungsanstalt - Kiel" (LUFA, independent laboratory). Certification avalaible.
- Water: tap water from Makrolon R drinking bottles, ad libitum
- Water quality assurance: water was collected twice yearly and analysed from analyses for chlorinated hydrocarbons, heavy metals and arsenic according to "Trinkwasserverordnung" by a governmental chemical institute for chemical quality. Water was controlled bacteriologically by governmental institute for public health. Certification available.
- Acclimation period: 7 days
- Justification for the choice of the species: standard experimental animal
- Animal identification: marked by numbered ear tags and individual cage cards (project number, test group, animal number, ear tag number, source, stain, sex, date of receipt)
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.5 °C ± 1.5 °C
- Humidity (%): 65 % ± 10 %
- Recording: by thermohygrograph
- Air changes (per hr): 16 times
- Photoperiod (hrs dark / hrs light): 12/24 from 7.00 a.m to 7.00 p.m.
- Intensity of light: 120 lux
Route of administration:
oral: feed
Vehicle:
other: basal diet ( incorporation in pellet)
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS DIET PREPARATION
- Rate of preparation of diet: the mixtures were prepared for the first 6 study weeks and again for the 6-13 weeks study phase.
- Mixing: test compound was weighted out for each concentration level and mixed with a small amount of basal diet (300 g), in a pulverized form. Mixing was continued in three steps to the final quantity. Each preparation was pelleted and identified.



Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
- Randomized collected samples from each batch and dose level were analysed for N-Oleyl Palmitamide concentrations (Identity, concentration, homogeneity and stability) by Dr. J.G.A. Kooyman, Clo AKZO Chemicals, Netherland BV, Research Centre Deventer, Analytical Department, Deventer.
- Results: the N-Oleyl Palmitamide concentrations were within a range of 10% of the target, homogeneity was within 10% of the average of Samples and stability was not deviating more than 7% of the originally determined average content.
- Basal diet (control) samples were analyzed to confirm the absence of N-Oleyl Palmitamide.
- Stability of the high concentration was tested after storage of the diet for 6 weeks and 4 months.
Duration of treatment / exposure:
13 weeks
Remarks:
Doses / Concentrations:
1200
Basis:
other: dietary concentration of N-Oleyl Palmitamide (ppm)
Remarks:
Doses / Concentrations:
6000
Basis:
other: dietary concentration of N-Oleyl Palmitamide (ppm)
Remarks:
Doses / Concentrations:
12000
Basis:
other: dietary concentration of N-Oleyl Palmitamide (ppm)
No. of animals per sex per dose:
40 male/40 female
4 test groups of 10 male and 10 female animals
Control animals:
yes, concurrent no treatment
Details on study design:
- Rationale for animal assignment: random, by a computerized randomization table (from both sexes)
Observations and examinations performed and frequency:
CLINICAL OBSERVATION
All animals were observed daily with regard to their Sensory and motor behaviour, hair coat, body orifices, urine and faecal excretion, for their general health status and dose responses. The observations were recorded daily and dose responses were summarized in weekly reports. Viability or mortality checks, resp., were performed twice daily.
SPECIAL CLINICAL EXAMINATION (by Dr. Trinks)
Prior to initiation, after 6 weeks and at termination:
- Ophthalmologic examinations: (cornea, episceral vessels, anterior chamber, pupil, lens, retina) using an ophthalmoscope
- The hearing, tested by simple noise production
- The reflex (awareness, emotion, coordination, autonomic functions)
In weekly intervals:
- Bodyweights, food consumption, water consumption
At the end of the test:
- Moribund animals/mortalities
- Complete macroscopic autopsy
- Organs observations
LABORATORY EXAMINATION (IBR laboratory)
After 6 weeks and at termination
- Hematologiacal and clinical chemistry investigations (samples from retrobulbar venous plexus)
- Urine collection (collected in 18 hours after 20 ml/Kg intragastric administration of water)
Sacrifice and pathology:
NECROPSY
All surviving animals were sacrificed at the end of the experiment by C02 asphyxiation.
A complete autopsy was performed for each animal, with the order of necropsy performed in a random manner (macropathological observations, registration of any abnormal findings)
DETERMINATION OF ORGAN WEIGHTS
Other examinations:
HISTOPATHOLOGICAL EXAMINATIONS (after organ fixation and tissue preparation)
Statistics:
Scheffè method for weight changes, food consumption and water consumption
Co-variance analysis and Scheffè method for organ weights
Variance analysis for dose-effect curves and Scheffè method for clinical chemistry and hematology
Results are expressed with the mean values per group and the standard deviation.
For bodyweights the percentage is calculated.
All control mean values are equalized with 100 %.
Clinical signs:
no effects observed
Description (incidence and severity):
Special reflex examinations after 6 weeks and at termination did not reveal any differences between control- and dose group animals
Mortality:
no mortality observed
Description (incidence):
Special reflex examinations after 6 weeks and at termination did not reveal any differences between control- and dose group animals
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Slight variations among groups were within a range of 10 % to the controls and, since there was no evidence of dose-relation
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
Description (incidence and severity):
Some sporadic differences were not dose-related and thus considered to be coincidental.
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
Occasionally occurring significant differences to the concurrent control group were not specifically dose-related
Urinalysis findings:
no effects observed
Description (incidence and severity):
The significant decrease of specific gravity in dose group III (males) after 6 weeks was not dose-related
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Description (incidence and severity):
Some incidental spontaneous changes were nearly equally distributed among control and dose group animals
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
Some spontaneous incidental lesions or lesions due to sacrifice were found
Histopathological findings: neoplastic:
not examined
Dose descriptor:
NOEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
other: dietary concentration
Sex:
male/female
Basis for effect level:
other: see 'Remark'
Dose descriptor:
NOEL
Effect level:
12 000 ppm
Based on:
other: dietary concentration
Sex:
male/female
Basis for effect level:
other: see 'Remark'
Critical effects observed:
not specified

The test material was incorporated into pelleted rat standard laboratory diet at concentrations of 1200 ppm (dose group I), 6000 ppm (dose group II) and 12000 ppm (dose group III), to perform a test according to OECD 408. 

These dietary concentrations were designed to provide daily dose levels of 100, 500, and 1000 mg/kg/day.

A concurrent control group received the basal diet only. Eachgroup consisted of 10 Wistar-rats per sex. The rats were exposed to the testcompound over an entire test period of 13 weeks. Analytical chemistry results indicated that the mixing procedure resulted in homogeneous mixtures of the test material in the diet. Periodic analyses during the study found the concentrations of test material in the diet to be within acceptable limits. A stability study conducted at the high concentration after storage for up to 4 months indicated that the test material was stabile in the diet. Under the experimental conditions of the study the test material had no adverseeffects on clinical signs, bodyweights, food consumption, water consumption,hematology, clinical chemistry, organs weights, and histopathology.

The "no-effect" - level of the test material was at a dietary concentrationof 12000 ppm, when fed to rats of the Wistar-strain over an entire test period of 13 weeks.

This no-adverse effect level correlates with a mean test compound intake of approximately 1000 mg/Kg/day.

 

Samples from all tissues listed below were removed from all animals; blocks and slides were prepared and examined for the control and high dose groups: skin, mammary gland, salivary gland, trachea, esophagus, thyroid (2x), parathyroid (2x), thymus, heart, lung, aorta, pituitary, tongue, liver, spleen, pancreas, kidney (2x), adrenal (2x), stomach, duodenum, jejunum, ileum, cecum, colon, seminal vesicle, lymph node (mesenteric+ cervical), ovary (2x)/ testis + epididymis (2x), prostate/uterus + vagina, urinary bladder, sciatic nerve, skeletal muscle, bone with marrow (sternum + femur), eye with N. opticus (2x), cerebrum with brain stem, cerebellum, spinal cord (2x), lacrimal gland (2x), macroscopic changes.

Microscopic evaluation of representative organ sections from all control and high dose animals did not reveal any test item related changes. Some spontaneous incidental lesions or lesions due to sacrifice were found evenly distributed among control and high dose animals of both sexes.

Conclusions:
The results of this study indicate, that the test material had no adverse effect on any of the tested parameters in rats of the Wistar-strain over the entire test period of 13 weeks. Thus a dietary concentration of 12000 ppm (high-dose group) revealed a clear "no-effect" level.
Significant intergroup differences in individual parameters were generally within normal limits and without any dose-relationship. These differences are therefore considered to be without biological importance and not attributable to the test material exposure.
Executive summary:

In an OECD 408 study, conducted according to GLP, the sub-chronic toxicity of the structurally similar read across substance in male and female Wistar rats a NOAEL of greater than or equal to 1,000 mg/kg/d was determined. The lack of test item related changes in the reproductive organs, accessory sexual organs, pituitary, adrenal gland, and thyroid provides evidence that the substance lacks the potential to cause reproductive toxicity and no treatment-related histopathological, haematological or clinical effects were observed.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
1

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 registration of the test material employs a combined approach for addressing the required endpoints. When data are not available, data from the read across substances are used to address the required endpoints.

 

The read across substances are structurally similar to registered substance. The substance and the proposed structural analogues are predominantly C32 – C36 alkyl fatty amides. These substances are expected to be metabolised by fatty acid amide hydrolase and esterases. Any differences among the substance structures are not expected to result in any significant differences in the toxicological effects or degradation products.

 

The following endpoints include supporting studies to provide evidence of read across suitability:

  • Repeated dose toxicity study (90 day)

 

Oral route:

In an OECD 408 study, conducted according to GLP, the sub-chronic toxicity of the structurally similar read across substance in male and female Wistar rats a NOAEL of greater than or equal to 1,000 mg/kg/d was determined. The lack of test item related changes in the reproductive organs, accessory sexual organs, pituitary, adrenal gland, and thyroid provides evidence that the substance lacks the potential to cause reproductive toxicity and no treatment-related histopathological, haematological or clinical effects were observed.

Inhalation route:

A short-term toxicity study does not need to be conducted because a reliable sub-chronic (90 days) or chronic toxicity study is available, conducted with an appropriate species, dosage, solvent and route of administration. This endpoint is waived.

Dermal route:

A short-term toxicity study does not need to be conducted because a reliable sub-chronic (90 days) or chronic toxicity study is available, conducted with an appropriate species, dosage, solvent and route of administration. This endpoint is waived.


Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
Study completed according to OECD Guideline under GLP conditions.

Justification for selection of repeated dose toxicity inhalation - systemic effects endpoint:
According to 8.6.1 Column 2 Annex VIII of Regulation (EC) 1907/2006 concerning the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH), the repeat dose toxicity, via inhalation route, of a substance does not need to be conducted if exposure of humans via inhalation is unlikely, taking into account the vapour pressure (for liquids) and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size. The test material is a solid block with no particles of an inhalable size (a granulometry study could not be conducted as the substance is a waxy solid block at room temperature) thus human exposure is considered unlikely, therefore, this endpoint is waived.

Justification for selection of repeated dose toxicity inhalation - local effects endpoint:
According to 8.6.1 Column 2 Annex VIII of Regulation (EC) 1907/2006 concerning the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH), the repeat dose toxicity, via dermal route, of a substance does not need to be conducted if exposure of humans via skin contact is unlikely, and the physicochemical and toxicological properties suggest no significant rate of absorption through the skin, for example, substances with a log Kow of <3 are more likely to penetrate the skin. The test material has a log Kow of 6.5 and therefore is unlikely to be absorbed via the skin. In addition, in an acute oral toxicity study the test material, showed no treatment- related effects (LD50 greater than 2,000 mg/kg bw; in a structurally similar substance read-across study, a 90-day repeated dose oral toxicity study showed no treatment-related effects (NOAEL 1,000 mg/kg bw/day) which indicates that no potential absorption occurs from the gastro-intestinal tract. In an acute dermal toxicity study, the read across substance showed no treatment related effects (LD50 greater than 2,000 mg/kg bw). Based upon the aforementioned information, human exposure is considered unlikely, and therefore, this endpoint is waived.

Justification for selection of repeated dose toxicity dermal - systemic effects endpoint:
According to 8.6.1 Column 2 Annex VIII of Regulation (EC) 1907/2006 concerning the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH), the repeat dose toxicity, via dermal route, of a substance does not need to be conducted if exposure of humans via skin contact is unlikely, and the physicochemical and toxicological properties suggest no significant rate of absorption through the skin, for example, substances with a log Kow of <3 are more likely to penetrate the skin. The test material has a log Kow of 6.5 and therefore is unlikely to be absorbed via the skin. In addition, in an acute oral toxicity study the test material, showed no treatment- related effects (LD50 greater than 2,000 mg/kg bw; in a structurally similar substance read-across study a 90-day repeated dose oral toxicity study showed no treatment-related effects (NOAEL 1,000 mg/kg bw/day) which indicates that no potential absorption occurs from the gastro-intestinal tract. In an acute dermal toxicity study, the read across substance showed no treatment related effects (LD50 greater than 2,000 mg/kg bw). Based upon the aforementioned information, human exposure is considered unlikely, and therefore, this endpoint is waived.

Justification for selection of repeated dose toxicity dermal - local effects endpoint:
According to 8.6.1 Column 2 Annex VIII of Regulation (EC) 1907/2006 concerning the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH), the repeat dose toxicity, via dermal route, of a substance does not need to be conducted if exposure of humans via skin contact is unlikely, and the physicochemical and toxicological properties suggest no significant rate of absorption through the skin, for example, substances with a log Kow of <3 are more likely to penetrate the skin. The test material has a log Kow of 6.5 and therefore is unlikely to be absorbed via the skin. In addition, in an acute oral toxicity study the test material, showed no treatment- related effects (LD50 greater than 2,000 mg/kg bw; in a structurally similar substance read-across study a 90-day repeated dose oral toxicity study showed no treatment-related effects (NOAEL 1,000 mg/kg bw/day) which indicates that no potential absorption occurs from the gastro-intestinal tract. In an acute dermal toxicity study, the read across substance showed no treatment related effects (LD50 greater than 2,000 mg/kg bw). Based upon the aforementioned information, human exposure is considered unlikely, and therefore, this endpoint is waived.

Justification for classification or non-classification

In an OECD 408 study, conducted according to GLP with the read-across substance the NOAEL(oral route)(rat) is greater than or equal to 1,000 mg/kg bw/d.

According to Regulation (EC) No 1272/2008 on the classification, labelling and packaging of substances and mixtures (CLP), a substance is classified as a Specific Target Organ Toxicity (repeated exposure) (STOT-RE) if the NOAEL is less than or equal to 100 mg/kg bw/d. Based upon the read across data, the test material is not classified for repeat dose toxicity.