Registration Dossier

Data platform availability banner - registered substances factsheets

Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Toxicological information

Repeated dose toxicity: oral

Currently viewing:

Administrative data

Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
06 June 1995 - 26 Sep 1995
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1997
Report date:
1997

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)
Version / remarks:
May 12, 1981
GLP compliance:
yes
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
cis-4-[3-(p-tert-butylphenyl)-2-methylpropyl]-2,6-dimethylmorpholine
EC Number:
266-719-9
EC Name:
cis-4-[3-(p-tert-butylphenyl)-2-methylpropyl]-2,6-dimethylmorpholine
Cas Number:
67564-91-4
Molecular formula:
C20H33NO
IUPAC Name:
(2R,6S)-4-[3-(4-tert-butylphenyl)-2-methylpropyl]-2,6-dimethylmorpholine

Test animals

Species:
rat
Strain:
Wistar
Details on species / strain selection:
Chbb: THOM (SPF)
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Dr. Karl Thomae GmbH, Biberach/Riss, FRG.
- Females (if applicable) nulliparous and non-pregnant: not specified
- Age at study initiation: 26 - 29 days
- Weight at study initiation: mean 194 g (male) / 147 g (female)
- Housing: single housing
- Diet (e.g. ad libitum): Kliba rats/mice/hamsters maintenance diet (343 meal, Klingentalmühle AG, Kaiseraugust, Switzerland), ad libitum
- Water (e.g. ad libitum): from water bottles, ad libitum
- Acclimation period: yes

DETAILS OF FOOD AND WATER QUALITY:
The food used in the study was assayed for chemical and microbiological contaminants.
The drinking water is regularly assayed for chemical contaminants as well as for the absence of microorganisms.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 24°C (air-conditioned)
- Humidity (%): 30 - 70%
- Air changes (per hr): not specified
- Photoperiod: 12 hrs dark / 12 hrs light

IN-LIFE DATES: From: June 06, 1995 To: Sep. 26, 1995

Administration / exposure

Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:
DIET PREPARATION
- Rate of preparation of diet (frequency): not specified
- Mixing appropriate amounts with (Type of food): Kliba rats/mice/hamsters maintenance diet (343 meal, Klingentalmühle AG, Kaiseraugust, Switzerland)
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The concentration control analyses yielded 128% -130% for the 1 ppm samples. Due to the small amount of the test substance and the determination limits of the analytical method, these values for the 1 ppm samples are assessed as being acceptable.
For the samples with nominal concentrations between 100 and 1,000 ppm, mean recoveries in the range of 93.3% - 99.9% were obtained thus confirming the theoretical contents.
Duration of treatment / exposure:
3 months
Frequency of treatment:
continuously
Doses / concentrationsopen allclose all
Dose / conc.:
1 ppm
Remarks:
corresponds to 0.1 mg/kg bw/d in males and females
Dose / conc.:
10 ppm
Remarks:
corresponds to 0.7 mg/kg bw/d in males, 0.8 mg/kg bw/d in females
Dose / conc.:
100 ppm
Remarks:
corresponds to 7.1 mg/kg bw/d in males, 8.5 mg/kg bw/d in females
Dose / conc.:
1 000 ppm
Remarks:
corresponds to 71.0 mg/kg bw/d in males, 77.7 mg/kg bw/d in females
No. of animals per sex per dose:
15 animals / sex in dose groups 0, 10, 100 and 1000 ppm
10 animals / sex in dose group 1 ppm
Control animals:
yes, plain diet
Details on study design:
- Dose selection rationale:
The 0, 1, 10, 100 and 1000 ppm dosages were selected on the basis of the results of the results of a 4-week repeated dose toxicity study and two 3-month feeding studies with the test substance where no animals died.
- Rationale for animal assignment (if not random): The animals were randomly assigned to the groups based upon body weight, separated by sex prior to the
first functional observational battery. The randomization list was drawn up by a computer.
- Fasting period before blood sampling for clinical biochemistry: yes, for about 16-20 hours


Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: on days -7, 22, 50 and 85

CLINICAL OBSERVATIONS: Yes
- Time schedule: Monday to Friday twice a day and Saturday, Sunday and public holidays once a day

BODY WEIGHT: Yes
- Time schedule for examinations: on day 0 and thereafter in weekly intervals

FOOD CONSUMPTION AND COMPOUND INTAKE:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: Yes

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: before the start and at the end of administration (study day 78 females and study day 80 males)
- Dose groups that were examined: each test group was examined before and the control and the high dose group were examined at the end of administration

HAEMATOLOGY: Yes
- Time schedule for collection of blood: on study day 91 (female) and 92 (male)
- Anaesthetic used for blood collection: No
- Animals fasted: Yes
- How many animals: 10 / dose group
- The following parameters were examined: leukocytes, erythrocytes, hemoglobin, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, platelets, prothrombin time

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: on study day 91 (female) and 92 (male)
- Animals fasted: Yes
- How many animals: 10 / dose
- The following parameters were examined: alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, serum cholinesterase. erythrocyte cholinesterase, brain cholinesterase, sodium, potassium, chloride, inorganic phosphate, calcium, urea, creatinine, glucose, total bilirubin, total protein, albumin, globulins, triglycerides, cholesterol, magnesium, gamma-glutamyltransferase

PLASMA/SERUM HORMONES/LIPIDS: Not specified

URINALYSIS: Yes
- Time schedule for collection of urine: on study day 87 (male) and 89 (female)
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes
- The following parameters were examined: volume, color, turbidity, nitrite, pH, protein, glucose, ketones, urobilinogen, bilirubin, blood, specific gravity, sediment

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: on study days -7, 22, 50 and 85
- Dose groups that were examined: all dose groups
- Battery of functions tested: sensory activity, grip strength, motor activity

IMMUNOLOGY: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes

HISTOPATHOLOGY: Yes

1. All gross lesions
2. Brain
3. Pituitary gland
4. Thyroid glands/ parathyroid glands
5. Thymus
6. Trachea
7. Lungs
8. Heart
9. Aorta
10. Salivary glands (mandibular gland, sublingual gland)
11. Liver
12. Spleen
13. Kidneys
14. Adrenal glands
15. Pancreas
16. Testes/ Ovaries
17. Uterus, oviducts, vagina
18. Epididymides, prostate, seminal vesicle
19. Skin
20. Esophagus
21. Stomach (fore- and glandular stomach)
22. Duodenum, jejunum, ileum
23. Cecum, colon, rectum
24. Urinary bladder
25. Lymph nodes (mesenteric and mandibular lymph node)
26. Female mammary gland
27. Skeletal muscle
28. Sciatic nerve
29. Sternum with sternal marrow
30. Bone marrow (femur)
31. Eyes
32. Femur with knee joint
33. Spinal cord (cervical, thoracic and lumbar cord)
34. Extraorbital lacrimal glands
Statistics:
Mean and standard deviations, the Kruskal-Wallis Test (two sided), Mann-Whitney U test (two-sided), Anova (two-sided), Fishers´s exact test and Dunnett´s test (two sided) were used.

Results and discussion

Results of examinations

Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
No substance related effects were obtained. The findings (loss of tail end in each one female of test groups 0 and 3) were spontaneous in nature.
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
In high dose (1,000 ppm ) males and females, body weight was statistically significantly impaired during the entire study. The impairment on day 91 in comparison to the control group was 14.7% in males and 12.2% in females. This was assessed as being substance-related. Corresponding body weight change
values were 24.9% and 25.3% below controls, respectively.

In 100 ppm females, body weight was statistically significantly impaired on days 21, 35, 42, 49, and 77; the values on day 91 was still 6.9% below control, although without statistical significance.
Nevertheless, this was assessed as being substance related. Corresponding body weight change values were 14.9% below controls.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
In high dose (1,000 ppm ) males and females, food consumption was statistically significantly impaired during the entire study. The impairment on day 91 in
comparison to the control group was 9.8% in males and 18.1% in females. This was assessed as being substance-related.
In the 10 ppm group males, statistically significantly increased values were seen on days 63, 77 and 84, and in the 100 ppm group females a statistically significantly decreased value was seen on day 21. Due to the isolated occurrence and the
lack of a dose-response relationship this was assessed as being incidental.
Food efficiency:
effects observed, treatment-related
Description (incidence and severity):
Food efficiency was statistically significantly impaired on single occasions in males and females treated with 1,000 ppm and in females treated with 100 ppm. This was assessed as being substance-related.
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
effects observed, non-treatment-related
Description (incidence and severity):
No substance-related findings were obtained. All findings (corneal stipplings, remainders of the pupillary membrane) were equally distributed between
the groups and thus spontaneous in nature.
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
At the end of the study slightly but statistically significantly decreased hemoglobin concentrations were found in the peripheral blood of the high dose females. The other hematological examinations and the clotting analyses did not reveal any substance-related changes.
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
After 3 months of test substance administration alanine aminotransferase activities were significantly increased in the sera of the high dose males and females. Moreover, marked decreases in serum cholinesterase activities were measured in test group 3 (68% of the control value) and 4 (69% of the control value) males and in test group 2 (71% of the control value), 3 (46% of the control value) and 4 (31% of the control value) females. The other serum enzymes as well as the erythrocyte and brain cholinesterase in the animals of either sex remained unaffected.
Blood chemistry examinations revealed increases in total bilirubin concentrations and decreases in triglyceride and cholesterol levels in the sera of the high dose males and females. However, the increase in total bilirubin seen in the high dose
males was not statistically significantly different to the corresponding control but appeared as a trend towards elevated concentrations.
Endocrine findings:
not examined
Urinalysis findings:
no effects observed
Behaviour (functional findings):
effects observed, treatment-related
Description (incidence and severity):
Open field observations:
In high, mid and low dose males rearing was statistically significantly decreased on day -7. This was clearly incidental in nature.
On day 22, each one male of test group 2 (10 ppm) and 3 (100 ppm) showed slight resistance against handling (rank 2) when removed from home cage. On day 50, 2 males of test group 2 and one male of test group 3 also showed slight resistance
against handling (rank 2). On day 85, one male of test group 2 was slightly apathetic when removing from home cage (rank 1) and one male showed slight resistance against handling (rank 2). However, as slight resistance against handling (rank 2) and "difficult to handle" (rank 3), was also observed sporadically in control animals, these findings were assessed as being incidental and not substance-related.

Sensorimotor tests / Reflexes:
The pupillary reflex test was retarded (rank 1) on day 22 in 1 high dose female, on day 50 in 8 high dose males and 5 high dose females, and on day 85 in 8 high
dose males and 5 high dose females.
Grip strength of forelimbs was statistically significantly reduced in high dose females on day 22.
The values of landing foot splay test were statistically significantly decreased in high dose males on day 50, in high dose females on days 22, 50 and 85 and in mid dose females day 22.
All these findings were assessed as being treatment-related.
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
In the male test animals, the relative organ weights of testes and brain of the high dose group and of the liver of the third dose group were statistically significantly increased (p <= 0.01, testes approximately 20%, brain approximately 20%, and liver
approximately 18%).
In the female test animals, the mean liver weights of the third and high dose group were statistically significantly increased (p <= 0.01, approximately 21% in the high dose group and approximately 13% in the third dose group).
Also the relative brain weights of the female third and high dose test animals were statistically significantly increased (p <= 0.01, approximately 13% in the high dose group and p <= 0.05, approximately 9% in the third dose group).
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Liver: A "Focus" was found in one male test animal of the second dose group.
Forestomach: The macroscopic finding "Margo plicatus thickened" was recorded for one female test animal of the control group.
Kidney: A macroscopically visible "Cyst" was diagnosed for one high dose female test animal.
Neuropathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
The mean absolute brain weight of the females in dose group 3 (100 ppm) showed a slight, significant decrease (p <= 0.05, approximately 6%).
The brain weight change is not seen to be of any biological significance.
A minimal (single, grade 1) axonal degeneration was found in one lumbar dorsal root of one male control. Minimal axonal degeneration was also found in the
tibial nerve of one male control and in the proximal sciatic nerve of another male control as well as in the proximal sciatic nerve of one high dose female and in the sural nerve of another high dose female.
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
For the macroscopic observation of a red brown "Focus" no histopathological correlate was found. All histopathological findings noted for the liver are seen to be spontaneous or incidental in nature.
The macroscopic finding "Margo plicatus thickened" of one female control test animal is caused by a focal, submucosal inflammation, histopathologically.
All histopathological findings noted for the forestomach are seen to be spontaneous or incidental in nature.
The macroscopic finding "Cyst" is verified by the histopathological examination.
All histopathological findings noted for the kidneys are seen to be spontaneous or incidental in nature.
Histopathological findings: neoplastic:
no effects observed

Effect levels

Dose descriptor:
NOAEL
Effect level:
10 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
behaviour (functional findings)
body weight and weight gain
clinical biochemistry
food consumption and compound intake
food efficiency
haematology
organ weights and organ / body weight ratios

Target system / organ toxicity

Key result
Critical effects observed:
not specified

Applicant's summary and conclusion