Registration Dossier

Toxicological information

Repeated dose toxicity: oral

Currently viewing:

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:
2010
Report Date:
2010

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents)
GLP compliance:
yes (incl. certificate)
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test substance: Oxooel 13
- Test substance No.: 08/0016-003
- Batch-Identification Tk253_20090901
- Purity: 18.5 area-% Mixture of isomers of tridecanol
7.5 area-% Mixture of isomers of tetradecane diol
71.2 area-% Mixture of isomers of ditridecane ether, tridecanoic acid tridecyl ester, tridecane tridecene ether and tetradecane diol tridecane ether
2.5 area-% Mixture of isomers of different trimeric components (ethers and esters)
- Homogeneity: given (visually)
- Stability: The stability was verified by reanalysis after completion of the study.
-Physical state/appearance: liquid /colorless, clear
- Storage conditions: ambient (RT), air sensitive (store under N2)

Test animals

Species:
rat
Strain:
other: Crl:WI(Han)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Sulzfeld, Germany
- Age at study initiation: 42 ± 1 days
- Weight at study initiation: males (mean): 185 g; females (mean): 135 g
- Housing: 5 animals per cage
- Diet (e.g. ad libitum): Kliba maintenance diet mouse/rat “GLP” (Provimi Kliba SA, Kaiseraugst, Switzerland), ad libitum
- Water (e.g. ad libitum): drinking water (from water bottles), available ad libitum
- Acclimation period: at least one week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 24
- Humidity (%): 30-70
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12 hours / 12 hours

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
olive oil
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS: Oxooel 13 was applied as a solution. To prepare this solution, the appropriate amount of test substance was weighed out depending on the desired concentration. Then, Olive Oil Ph. Eur. was filled up to the desired volume, subsequently released manually. The test-substance preparations were produced once a week.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The analyses of the test-substance preparations have been carried out at Competence Center Analytics, BASF SE, Ludwigshafen, Germany as a separate study under the responsibility of a study director of this testing facility. The study was carried out in compliance with the Principles of Good Laboratory Practice.
Method: High temperature capillary GC with area-% evaluation and correction with water content
Sample preperation: The test item was injected as it is.
Results: The area-% evaluation after correction with water content leads to the following results: Group 1: 18.5 area-%; Group 2: 7.5 area-%; Group 3: 71.2 area-%; Group 4: 2.5 area-%. The results are the mean out of two independent determinations. The different components were grouped according to: Group 1: Mixture of isomers of tridecanol; Group 2: Mixture of isomers of tetradecane diol; Group 3: Mixture of isomers of ditridecane ether, tridecanoic acid tridecyl ester, tridecane tridecene ether and tetradecane diol tridecane ether; Group 4: Mixture of isomers of different trimeric components (ethers and esters)
Duration of treatment / exposure:
28 days
Frequency of treatment:
once daily
Doses / concentrations
Remarks:
Doses / Concentrations:
0, 100, 300 and 1000 mg/kg bw/d
Basis:
actual ingested
No. of animals per sex per dose:
5 animals
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: A test study at dose levels of 0, 300 and 1000 mg/kg body weight/day for 2 weeks was performed (experimental conduct in accordance with GLP, without a GLP-status). The only treatment-related finding that was obtained during the study was salivation after treatment in rats of both test groups.
Positive control:
no

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily on working days and once daily on Saturdays, Sundays and public holidays
- Cage side observations: mortality

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: prior to the administration period and thereafter at weekly intervals
- Parameters examined: abnormal behavior during “handling”, fur, skin, posture, salivation, respiration, activity/arousal level, tremors, convulsions, abnormal movements, impairment of gait, lacrimation, palpebral closure, exophthalmus, feces (appearance/ consistency), urine, pupil size

BODY WEIGHT: Yes
- Time schedule for examinations: During the administration period the body weight was determined on day 0 (start of the administration period) and thereafter at weekly intervals

FOOD CONSUMPTION: Yes
- Time schedule: weekly over a period of 1 day
- Food consumption calculated as mean food consumption in grams per rat and day.

WATER CONSUMPTION: Yes
- Time schedule for examinations: daily

HAEMATOLOGY: Yes
- Time schedule for collection of blood: in the morning at the end of the study
- Anaesthetic used for blood collection: Yes, isoflurane
- How many animals: 5 animals per test group and sex
- Parameters examined: Leukocyte count (WBC), Erythrocyte count(RBC), Hemoglobin (HGB), Hematocrit (HCT), Mean corpuscular volume (MCV), Mean corpuscular hemoglobin (MCH), Mean corpuscular hemoglobin concentration (MCHC), Platelet count (PLT), Differential blood count, Reticulocytes, Prothrombin time (Hepato Quick’s test; HQT)

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: in the morning at the end of the study
- How many animals: 5 animals per test group and sex
- Parameters examined: Alanine aminotransferase, Aspartate aminotransferase, Alkaline phosphatase, γ-Glutamyltransferase, Sodium, Potassium, Chloride, Inorganic phosphate, Calcium, Urea, Creatinine, Glucose, Total bilirubin, Total protein, Albumin, Globulins, Triglycerides, Cholesterol, Magnesium, Bile acids

URINALYSIS: Yes
- Time schedule for collection of urine: urine was collected overnight
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes, withdrawal of food and water overnight
- Parameters examined: pH, protein, Glucose, Ketones, Urobilinogen, Bilirubin, Blood, Specific gravity, Sediment, Color, turbidity, Volume

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: at the end of the administration period
- Dose groups that were examined: in all rats
- Battery of functions tested: home cage observations (posture, tremor, convulsions, abnormal movements, impairment of gait), open field observations (behavior when removed from cage, fur, skin, salivation, nose discharge, lacrimation, eyes/ pupil size, posture, palpebral closure, respiration, tremors, convulsions, abnormal movements, impairment of gait, activity/ arousal level, feces (number of fecal pellets/ appearance/ consistency) within two minutes, urine (appearance/ quantity) within two minutes, number of rearings within two minutes.), sensorimotor tests/reflexes (approach response, touch response, vision ("visual placing response"), pupillary reflex, pinna reflex, audition ("startle response"), coordination of movements ("righting response"), behavior during "handling", vocalization, pain perception ("tail pinch"), grip strength of forelimbs, grip strength of hindlimbs, landing foot-splay test), motor activity
Sacrifice and pathology:
GROSS PATHOLOGY: Yes, Organ weights,Organ/tissue fixation
HISTOPATHOLOGY: Yes, All gross lesions, Adrenal glands, Bone marrow (femur), Brain, Cecum, Cervix, Colon, Duodenum, Epididymides, Eyes, Heart, Ileum, Jejunum (with Peyer’s patches), Kidneys, Liver, Lung, Lymph nodes (mesenteric and axillary lymph nodes), Ovaries, Pituitary gland, Prostate, Rectum, Sciatic nerve, Spinal cord (cervical, thoracic and lumbar cords), Spleen, Seminal vesicle with coagulation glands, Skeletal muscle, Sternum with marrow, Stomach (forestomach and glandular stomach), Testes, Thymus, Thyroid glands (with parathyroid glands), Trachea, Urinary bladder, Uterus, Vagina
Statistics:
Body weight, body weight change: A comparison of each group with the control group was performed using DUNNETT's test (two-sided) for the hypothesis of equal means.
Feces, rearing, grip strength forelimbs, grip strength hindlimbs, foot-splay test, motor activity: Non-parametric one-way analysis using KRUSKALWALLIS test (two-sided). If the resulting p-value was equal or less than 0.05, a pairwise comparison of each dose group with the control group was performed using WILCOXON test (two-sided) for the equal medians.
Clinical pathology parameters, urine volume, urine specific gravity: Non-parametric one-way analysis using KRUSKAL-WALLIS test (two-sided). If the resulting p-value was equal or less than 0.05, a pairwise comparison of each dose group with the control group was performed using WILCOXON-test (two-sided) for the equal medians.
Urinalysis, except color, turbidity, volume and specific gravity: Pairwise comparison of each dose group with the control group using FISHER's exact test for the hypothesis of equal proportions.
Weight parameters: Non-parametric one-way analysis using KRUSKAL-WALLIS test (two-sided). If the resulting pvalue was equal or less than 0.05, a pairwise comparison of each dose group with the control group was performed using WILCOXON-test (two-sided) for the equal medians.

Results and discussion

Results of examinations

Details on results:
CLINICAL SIGNS AND MORTALITY: No rat died prematurely in the present study.
Slight and moderate salivation after treatment was seen in all males and 4 females of test group 1000 mg/kg bw/d. This finding was observed from day 3 onwards on several days of the study. Slight salivation after treatment was seen in 2 male and 2 female rats of test group 300 mg/kg bw/d from day 5 onwards each on one or two several days as well as in 1 male of test group 100 mg/kg bw/d on day 25.

FOOD CONSUMPTION: No test substance-related effects on food consumption were obtained.

WATER CONSUMPTION: No test substance-related findings were observed.

BODY WEIGHT: No test substance-related changes of body weight and body weight change were observed in any test group. Body weight change was significantly increased in female animals of test group 1000 mg/kg bw/d on day 14 (+26 %). These changes were assessed as being incidental and not related to treatment.

HAEMATOLOGY: At the end of the study, in males of test group 1000 mg/kg bw/d the total white blood cell (WBC) counts were lower compared to controls, although not significant. This was due to a decrease of the absolute lymphocyte counts. Consequently, the relative neutrophil counts were increased.
Lower absolute counts of the large unstained cells (LUC) might be an artifact, because some activated lymphocytes normally were counted as LUCs and, therefore, a decrease of absolute lymphocyte counts might have lead to a corresponding decrease of LUC counts. No pathological effect could be correlated with a decrease of LUC counts. Therefore, the decrease of LUC counts was not regarded as a treatment-related effect.
In males of test group 1000 mg/kg bw/d, the red blood cell (RBC) counts were marginally increased. This was the only changed red blood cell parameter and, therefore, the alteration was regarded as possibly treatment-related.
In females of test group 100 mg/kg bw/d the hematocrit values were slightly increased. This was the only altered red blood cell parameter in female animals, and the values were not dose-dependently changed. Therefore, this alteration was regarded as incidental and not treatment-related.

CLINICAL CHEMISTRY: No treatment-related, adverse changes among clinical chemistry parameters were measured.
In males of test groups 300 and 1000 mg/kg bw/d the total bilirubin values were lower compared to controls. This was most probably due to an increased conjugation of bilirubin in the liver followed by an increased excretion via the bile and the kidneys. No pathological finding could be correlated with a lower bilirubin concentration. Therefore, this change was regarded as an adaptive and non-adverse effect.
In females of test group 1000 mg/kg bw/d the alanine aminotransferase (ALT) activities were moderately lower compared to controls. This alteration corresponded to the increased relative liver weights in females of test groups 300 and 1000 mg/kg bw/d without any histopathological correlate. Additionally, in females of test groups 300 and 1000 mg/kg bw/d the serum bile acid levels were lower compared to controls, which became statistically significant in test group 1000 mg/kg bw/d. The conjugation rate of bile acids in liver cells was higher and subsequently more bile acids were excreted via bile. The active re-absorption capacity of bile acids in the ileum was exceeded and therefore bile acids were excreted in the feces. In summary, these effects were most probably due to a compound-induced microsomal enzyme induction in liver cells and were therefore regarded as an adaptive and not an adverse effect (PSD Guidance Document, 2007).

URINALYSIS: No treatment-related changes among urinalyses parameters were measured.
In males of test groups 300 and 1000 mg/kg bw/d higher incidences of granulated and epithelial cell casts were found in the urine sediment (not significant in test group 1000 mg/kg bw/d). This was the only alteration regarding kidney markers.
As shown in the pathohistological investigation these findings were most probably due to the “inducible α2μ globulin nephropathy syndrome” which is described as species and gender specific effect in male rats (G. C. Hard et al., 1993), and is therefore not relevant as adverse effect for humans.

NEUROBEHAVIOUR: Deviations from "zero values" were obtained in several rats. However, as most findings were equally distributed between test substance-treated groups and controls, were without a doseresponse relationship or occurred in single rats only, these observations were considered to
have been incidental.

ORGAN WEIGHTS:
- Absolute weights: When compared to the control group (set to 100 %), the mean absolute weights of following organs were significantly increased: Thyroid glands: in males dose group 100 mg/kg bw/d: 149 %*, in females dose group 300 mg/kg bw/d: 131 %* (* : p ≤ 0.05). All other mean absolute weight parameters did not show significant differences when compared with the control group.
- Relative organ weights: When compared to the control group (set to 100 %), the mean relative weights of following organs were significantly increased: Thyroid glands: in males dose group 100 mg/kg bw/d: 151 %*, in females dose group 300 mg/kg bw/d: 129 %* (* : p ≤ 0.05); Liver: in females dose group 300 mg/kg bw/d: 109 %*, dose group 1000 mg/kg bw/d: 114 %** (* : p ≤ 0.05, **: p ≤ 0.01). All other mean relative weight parameters did not show significant differences when compared with the control group.
For the increased mean relative liver weights in females of test groups 300 mg/kg bw/d and 1000 mg/kg bw/d a treatment-related effect cannot be ruled out.
Due to the lack of dose-response relationship, the increase of the mean absolute and relative thyroid weights in males of test group 100 mg/kg bw/d as well as in females of test group 300 mg/kg bw/d was considered incidental.

GROSS PATHOLOGY: There were no gross lesions in treated male and female rats.

HISTOPATHOLOGY:
- Kidneys: A various number (very few to a moderate number) of eosinophilic droplets were observed in the cytoplasm of proximal tubules in all male animals. The eosinophilic droplets stained red with Mallory Heidenhain. The severity was slightly increased in males of test group 1000 mg/kg bw/d.
-Liver: There were no histopathological correlates for the increased mean relative liver weights in females of test groups 300 and 1000 mg/kg bw/d.
- Prostate: Very few or few multifocal lymphoid infiltrates were observed in the prostate of 2 male animals in test group 1000 mg/kg bw/d. Lymphoid infiltrates are known as background lesion and are often seen in the prostate also in controls. Therefore, their occurrence was considered incidental.
All other findings noted were either single observations or they were biologically equally distributed between control and treatment groups. All of them were considered to be incidental or spontaneous in origin and without any relation to treatment.

Effect levels

open allclose all
Dose descriptor:
NOAEL
Effect level:
300 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male
Basis for effect level:
other: see 'Remark'
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
female
Basis for effect level:
other: No signs of toxicity. (Salivation was considered to be related to either the bad taste of the test substance or local affection of the upper digestive tract. This finding was not considered to be an adverse and toxicologically relevant effect.)

Target system / organ toxicity

Critical effects observed:
not specified

Applicant's summary and conclusion