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Toxicological information

Repeated dose toxicity: oral

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Administrative data

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2003

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)
Version / remarks:
adopted July 27th, 1995
Qualifier:
according to guideline
Guideline:
EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))
Version / remarks:
adopted July 30th, 1996
Qualifier:
according to guideline
Guideline:
other: U.S. EPA: OPPTS 870.3050, Health Effects Test Guidelines: Repeated dose 28-day oral toxicity study in rodents, July 2000
GLP compliance:
yes
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
(Z)-octadec-9-enylamine
EC Number:
204-015-5
EC Name:
(Z)-octadec-9-enylamine
Cas Number:
112-90-3
Molecular formula:
C18H37N
IUPAC Name:
octadec-9-en-1-amine
Details on test material:
- Name of test material (as cited in study report): Genamin OL 100 D
- Physical state: white paste
- Analytical purity: app. 100%
- Purity test date: 2002-08-16
- Lot/batch No.: DEGE 905 602
- Expiration date of the lot/batch: 2003-07-12
- Stability under test conditions: guaranteed for 4h in sesame oil by the sponsor
- Storage condition of test material: room temperature in fume cupboard

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Harlan Winkelmann GmbH, Borchen
- Age at study initiation: app. 6 weeks
- Weight at study initiation: app. 126g (males), 123g (females)
- Fasting period before study: no
- Housing: 5 animals per cage in Makrolon type IV cages with soft wood granulate
- Diet (e.g. ad libitum): "ssniff" ad lib., except for the period in which the animals were kept in diuresis cages
- Water (e.g. ad libitum): tap water ad lib. except for the period in which the animals were kept in diuresis cages
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3°C
- Humidity (%): 50 +/- 20%
- Photoperiod (hrs dark / hrs light): 12h/12h

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: sesame oil
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS: The test substance was dissolved daily in sesamy oil.

VEHICLE
- Justification for use and choice of vehicle (if other than water): solubility
- Concentration in vehicle: 0.325 - 5 mg/mL
Volume of vehicle + test substance: 10ml/kg b.w.
Duration of treatment / exposure:
28days
Frequency of treatment:
daily
Doses / concentrations
Remarks:
Doses / Concentrations:
3.25, 12.5, 50mg/kg b.w.
Basis:
actual ingested
No. of animals per sex per dose:
5 (+ 5 per sex per recovery group)
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale:
In a dose range finding study, 3 males and 3 females received 25, 100, and 400mg/kg of the test substance for 14 days. One high dose male and female died on days 4 and 7, respectively. The other animals of this dose group were killed for animal welfare reasons. Necropsy revealed changes in the the stomach and intestinal mucosa, probably due to the irritant properties of the test substance. Mid dose animals showed impairments of motility and respiration, with males being clearly more sensitive. Body weight gain was also impaired, and necropsy revealed reddening of the stomach mucosa. Except for one female, that showed uncoordinated gait on study day 2 only, no signs of toxicity were observed in the low dose group.
Based on the results, dose levels of 3.25, 12.50 and 50.00 mg/kg body weight per day were selected for the 4-week repeat dose gavage study.

- Rationale for selecting satellite groups: random
- Post-exposure recovery period in satellite groups: 14 days
Positive control:
none

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily
- Cage side observations: mortality, clinical signs

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: weekly
Changes in skin, fur, eyes, mucous membranes, occurrence of secretions and excretions and autonomic activity such as lacrimation, salivation, nasal discharge, piloerection, pupil size, and unusual respiratory pattern. Changes in gait, posture, and response to handling as well as the presence of clonic or tonic movements, tremor, and any other abnormal motor movements (such as excessive grooming, repetitive circling or other stereotypes) or bizarre behavior (e.g. self-mutilation, walking backwards) were also recorded. In addition, defecation and urination were evaluated.

BODY WEIGHT: Yes
- Time schedule for examinations: twice weekly

FOOD CONSUMPTION:
- evaluated twice weekly

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: during detailed clinical examinations
- Dose groups that were examined: all

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at study termination
- Anaesthetic used for blood collection: Yes (Ketamine-Hydrochloride + Xylazine)
- Animals fasted: No
- How many animals: all
- The following parameters were examined: erythrocyte count, mean corpuscular hemoglobin, mean corpuscular volume, mean corpuscular hemoblobin concentration, reticulocyte counts, hemoglobin, hematocrit, Heinz Body counts (only high dose and control), differential leukocyte counts, clotting time, thrombocyte counts

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at study termination
- Animals fasted: No
- How many animals: all
- The following parameters were examined: GGT, ALAT, albumin, albumin/globulin ratio, alkaline phosphatase, ASAT, bilirubin, Ca, Cl, cholesterol, creatinine, globulin, glucose, inorganic phosphorous, K, Na, total protein, triglycerides, urea, uric acid

URINALYSIS: Yes
- Time schedule for collection of urine: overnight from days 25 to 26
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes
- The following parameters were examined: appearance, bilirubin, blood, color, glucose, ketone bodies, microscopic examination (control and high dose only), pH, protein, urobilinogen, specific weight

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: at study termination
- Dose groups that were examined: all
- Battery of functions tested: sensory reactivity incl. startle response, response to approach, righting reflex, pupillary constriction, motor activity, forelimb and hindlimb grip strength
Sacrifice and pathology:
GROSS PATHOLOGY: Yes: skin, orifices, eyes, teeth, oral mucosa, internal organs
HISTOPATHOLOGY: Yes
organ weights: adrenals, heart, spleen, testes, kidneys, brain, epididymides, liver, thymus
organs processed for histpathological investigations: adrenals, lung, stomach, bone marrow, lymph nodes (mandibular, iliac), testes, thymus, brain, epididymides, sciatic nere, thyroid gland, heart, ovaries trachea, small and large intestine, prostate, seminal vesicle, urinary bladder, uterus, kidneys, spinal cord, nasal cavities, liver, spleen, all gross lesions.

The lungs (intratracheally) and the nasoturbinates (in skull) were instilled/infused with formaldehyde (4 %). Following completion of the fixation, the lungs and nasoturbinates were fixed, together with the other organs, in formalin solution (4% neutral buffered formaldehyde).

Results and discussion

Results of examinations

Details on results:
CLINICAL SIGNS AND MORTALITY
There were no unscheduled deaths throughout the study.
Clinical observations remained unaffected by the administration of the test compound in the low and intermediate dose groups.
In the high-dose group impairments of motility from the 2nd or 3rd week onwards (stilted or uncoordinated gait) were observed in 2 males and 1 female of the main group animals up to the end of the treatment period. These observations were also noted in 4 females in the high dose recovery group starting at the end of treatment (2nd or 3rd week), with subsequent recovery during the recovery period. Respiratory sounds were noted in one female of the high-dose group only on day 13 of the study.
No changes of the oral mucosa, or impairment of dental growth was observed in all groups.

BODY WEIGHT AND WEIGHT GAIN
Body weight gain was not influenced by treatment in low dose animals and mid dose females. The mean body weight was significantly decreased in high dose males and females (app. -10% for both genders) at the end of treatment, with clear subsequent recovery. Mean body weight was slightly but statistically significantly decreased for mid-dose males from study day 22 onwards until end of treatment. This finding was marginal, i.e., less than 5 % as compared to the control. Moreover, it was not observed for the mid-dose females. Hence, it was considered not toxicologically significant.

FOOD CONSUMPTION
unaffected by test substance

OPHTHALMOSCOPIC EXAMINATION
No opacity of the refracting media of the eyes was observed in all groups.

HAEMATOLOGY
- Slightly increased hematocrit values and slightly decreased reticulocyte counts for high-dose males (a similar tendency without statistical significance was noted for high-dose females), subsequent recovery
- Slightly increased white blood cell (WBC) counts for high-dose males and females (not statistically significant, however, for males outside the range of the historical control data), with a shift towards increased neutrophils in both genders, with subsequent recovery

CLINICAL CHEMISTRY
All observed differences to control animals were either within historical control data or showed no dose response relationship.

URINALYSIS
unaffected by test substance

NEUROBEHAVIOUR
Neurological measurements were unaffected by treatment.

ORGAN WEIGHTS
unaffected by test substance

GROSS PATHOLOGY
There were no gross pathology findings, which could be related to administration of the test compound.

HISTOPATHOLOGY: NON-NEOPLASTIC
There were no histopathological findings, which could be related to administration of the test compound.

Effect levels

Dose descriptor:
NOAEL
Effect level:
12.5 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: reduced body weight

Target system / organ toxicity

Critical effects observed:
not specified

Applicant's summary and conclusion

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