Registration Dossier

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other: GLP compliant guideline study, available as unpublished report, no restrictions, fully adequate for assessment.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2007
Report Date:
2007

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents)
Qualifier:
according to
Guideline:
EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))
GLP compliance:
yes
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): 1,1,1,2,2,3,3,4,4,5,5,6,6-Tridecafluorooctane
- Physical state: colourless, transparent liquid
- Batch number: #060616
- Purity: 99.9%
- Storage condition of test material: at room temperature
- Stability under storage conditions: stable

Test animals

Species:
rat
Strain:
other: Crl:CD(SD)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Japan, Inc. (Hino Breeding Center; 735; Shimokomatsuki, Hino-cho, Gamo-gun, Shiga 529-1633, Japan)
- Age at study initiation: 5 weeks
- Weight at study initiation: 117.5-143.4 g (males), 109.2-128.5 g (females)
- Fasting period before study: no
- Housing: hanging stainless steel cages with wire-mesh floor at 1 animal/cage (165 Wx300 Dx150 H mm, TOKIWA KAGAKU KIKAI)
- Diet: MF pelleted diet (lot no. 060601 and 060802, Oriental Yeast), ad libitum
- Water: chlorinated water from the Hita City supply via automatic watering system with sipper tubes, ad libitum
- Acclimation period: 7 days (including 6 days quarantine)

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22.7-23.8
- Humidity (%): 48.5-65.1
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12 / 12

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
olive oil
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:
The test substance was accurately weighed and mixed with olive oil to prepare 2.0 w/v% formulation. The lower concentrations of 0.08 and 0.4 w/v% were diluted from 2.0 w/v%. These were prepared twice a week. The formulations were stored in a dark and cold place.

VEHICLE
- Justification for use and choice of vehicle: The test substance was not dissolved in purified water and olive oil, and the condition of the olive oil suspension was good. Therefore, olive oil (Fujimi Pharmaceutical) was used as the vehicle.
- Concentration in vehicle: 0.08, 0.4 and 2.0 w/v%
- Amount of vehicle (if gavage): 10 ml/kg bw
- Lot/batch no.: 038OHS
Analytical verification of doses or concentrations:
yes
Duration of treatment / exposure:
28 days
Frequency of treatment:
once daily
Doses / concentrations
Remarks:
Doses / Concentrations:
8, 40, 200 mg/kg bw/day
Basis:
actual ingested
No. of animals per sex per dose:
5
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: 7-day oral repeated dose toxicity study at 4 doses (25, 250, 500 and 1000 mg/kg bw/day)
- 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: three times a day during the dosing period, once daily during the recovery period

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: once weekly

BODY WEIGHT: Yes
- Time schedule for examinations: before dosing, and on days 1, 3, 8, 12, 17, 21, 26 and 28 during the dosing period; on days 1, 5, 10 and 14 during the recovery period

FOOD CONSUMPTION:
- Time schedule: before dosing, on days 1, 3, 8, 15, 22 and 28 during the dosing period; on days 1, 4, 8 and 14 during the recovery period

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at end of dosing period and at end of recovery period
- Anaesthetic used for blood collection: Yes (ether)
- Animals fasted: Yes
- How many animals: all animals of the respective groups
- Parameters examined were red blood cell count (RBC), white blood cell count (WBC), hemoglobin concentration (Hb), hematocrit value (Ht), mean corposcular volume (MCV), mean corposcular hemoglobin (MCH), mean corposcular hemoglobin concentration (MCHC), platelet count, reticulocyte count, prothrombin time (PT), activated partial thromboplastin time (APTT), differentiation of leukocytes (neutrophils, eosinophils, basophils, lymphocytes, monocytes, large unstained cells (LUC).

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at end of dosing period and at end of recovery period
- Animals fasted: Yes
- How many animals: all animals of the respective groups
- Parameters examined were aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), cholinesterase (ChE), γ-glutamyl transpeptidase (γ-GTP), total cholesterol (T-Cho), triglyceride (TG), glucose, total protein, albumin, A/G ratio, blood urea nitrogen (BUN), creatinine, total bilirubin (T-Bil), calcium, inorganic phosphorous (IP), sodium, potassium, chloride.

URINALYSIS: Yes
- Time schedule for collection of urine: at end of dosing period and at end of recovery period
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes
- Parameters examined were urine volume, colour, turbidity, urine specific gravity, pH, protein, glucose, occult blood, urinary sediments.

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: week 4
- Dose groups that were examined: all animals
- Battery of functions tested: sensory activity, grip strength, motor activity
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
The weights of the following organs were measured in all animals: liver, heart, kidneys, testes, epididymides, ovaries, brain, spleen thymus, adrenals. The relative organ weight was also calculated based on the body weight at the time of necropsy.

HISTOPATHOLOGY: Yes
Organs and tissues examined were trachea, lungs, incisors, stomach, intestine (duodenum to rectum, with Peyer's patches), liver, heart, kidneys, urinary bladder, testes, epididymides, prostate, seminal vesicle, ovaries, uterus, vagina, brain (cerebrum, cerebellum and pons), spinal cord, sciatic nerve, bone marrow (femur), axillar and mesenteric lymph nodes, spleen, thymus, pituitary gland, thyroids (with parathyroids), adrenals, eye ball.
Other examinations:
none
Statistics:
Data regarding body weights (excluding those at the time of necropsy) food intakes, hematological examinations, blood chemical examinations, urine volume and specific gravity, organ weights, grip strength and locomotor activity count were analyzed using the Bartlett's test for homogeneity of variance. If the variances were homogeneous at a significance level of 5%, one way analysis of variance was performed. If there was a significant difference in this analysis, the difference between the vehicle control group and each of the treatment groups was analyzed by the Dunnett's test.
If the variances were not homogeneous, the Kruskal-Wallis's test was used. If there was a significant difference in this analysis, the difference between the vehicle control group and each of the treatment group was analyzed by the nonparametric Dunnett's test.
FOB numerical data was analyzed using the Kruskal-Wallis's test. If there was a significant difference in this analysis, the difference between the vehicle control group and each of the treatment groups was analyzed by the nonparametric Duenna's test.

Results and discussion

Results of examinations

Details on results:
CLINICAL SIGNS
During Dosing Period
Males: Salivation only after administration was observed in one animal of the 8 mg/kg group, two animals of the 40 mg/kg group and six animals of the 200 mg/kg group.
Females: Salivation only after administration was observed in two animals of the 40 mg/kg group and seven animals of the 200 mg/kg group.
During Recovery Period
Males: Mottled teeth were observed in three animals of the 200 mg/kg group.
Females: No abnormalities were noted.

DETAILED OBSERVATIONS
During Dosing Period
No abnormalities were noted in all groups.
During Recovery Period
No abnormalities were noted in all groups.

SENSORIMOTOR FUNCTION
During Dosing Period
No abnormalities were noted in all groups.
During Recovery Period
Not examined.

BODY WEIGIITS
During Dosing Period
No abnormalities were noted in all groups.

During Recovery Period
No abnormalities were noted in all groups.

FOOD INTAKES
During Dosing Period
Males: No abnormalities were noted.
Females: Food intakes were decreased in the 40 and 200 mg/kg groups on day 22.
During Recovery Period
No abnormalities were noted in all groups,

HEMATOLOGICÄL EXAMINATIONS
At Termination of Dosing Period
Males: No abnormalities were noted.
Females: WBC was increased in the 8 mg/kg group.
At Termination of Recovery Period
Males: MCV was decreased in the 200 mg/kg group.
Females: Eosinophils were decreased in the 200 mg/kg group.

BLOOD CHEMICAL EXAMINATIONS
At Termination of Dosing Period
Males: Total cholesterol and albumin were decreased in the 200 mg/kg group.
Females: Cholinesterase was decreased in the 40 and 200 mg/kg groups.
At Termination of Recovery Period
Males: No abnormalities were noted.
Females: Total cholesterol was decreased in the 200 mg/kg group.

URINALYSES
At Termination of Dosing Period
No abnormalities were noted in all groups.
At Termination of Recovery Period
No abnormalities were noted in all groups.

ORGAN WEIGHTS
At Termination of Dosing Period
Males: Relative liver and kidney weights were increased in the 200 mg/kg group.
Females: Relative liver weight was increased in the 200 mg/kg group.
At Termination of Recovery Period
Males: No abnormalities were noted.
Females: Absolute and relative kidney weights and relative ovary weights were increased in the 200 mg/kg group.

NECROPSY
At Termination of Dosing Period
Males: Enlargement of the liver was observed in two animals (nos. 22 and 25) of the 200 mg/kg group.
Females: A decrease in size of the left lobe in the thyroid was observed in one animal (no. 43) of the 8 mg/kg group. Enlargement of the liver in two animals (nos. 53 and 55) and pelvic dilatation in the kidneys in one animal (no. 53) were observed in the 200 mg/kg group.
At Termination of Recovery Period
Males: Mottled teeth were observed in three animals (nos. 26, 29 and 30) were observed in the 200 mg/kg group.
Females: No abnormalities were noted.

HISTOPATHOLOGICAL EXAMINATIONS
At Termination of Dosing Period
Males: Basophilic tubules in the kidney in one animal (no. 4) and round cell infiltration in the prostate in one animal (no. 5) were observed in the vehicle control group.
Centrilobular hypertrophy of the hepatocytes in the liver in four animals (nos. 21, 22, 24, 25), solitary cyst in the medulla in one animal (no. 23) and subcupsular solitary cyst in one animal (no. 21) in the kidney were observed in the 200 mg/kg group.
Females: Mineralization in the corticomedullary junction of the kidney in two animals (nos. 31 and 32) and ultimobronchial rest in the thyroid in one animal (no. 33) were observed in the vehicle control group. Aplasia of the left lobe in the thyroid in one animal (no. 43) was observed in the 8 mg/kg group. Pelvic dilatation in the kidney was observed in one animal (no. 53) of the 200 mg/kg group.
At Termination of Recovery Period
Males: No abnormalities were noted.
Females: Solitary cyst in the medulla of the kidney was observed in one animal (no. 38) of the vehicle control group.

Effect levels

Dose descriptor:
NOAEL
Effect level:
40 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: increased relative liver weights and enlargement of the liver in males and females and centrilobular hypertrophy of the hepatocytes and increased relative kidney weights in males of the 200 mg/kg bw/day groups

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

Discussion of the results

No death occurred in the dosing and recovery periods.

Salivation was transiently observed in males of all treatment groups and females of the 40 and 200 mg/kg bw groups; however, it was not considered to be toxicologically significant since animals salivated just after dosing and there were no related changes to this.

Food consumption was decreased in females of the 40 and 200 mg/kg groups on day 22 of the dosing period. This was transient and not dose-related, and there were not body weight changes related to the decreased food intake on the same day. Therefore this change was not considered to be treatment-related.

In blood chemistry, decreased cholinesterase in females of the 40 and 200 mg/kg groups and decreased total cholesterol and albumin in males of the 200 mg/kg group, were noted, and these were not considered related to the test article, since they were slight changes within the background value of the laboratory and lesion of the liver in males were severer than that in females.

In organ weights, relative liver weights were increased in males and females of the 200 mg/kg bw/day groups. In necropsy, enlargement of the liver was observed in males and females of the 200 mg/kg bw/day groups. In histopathological examinations, centrilobular hypertrophy of the hepatocytes was observed in males of the 200 mg/kg bw/day group.

Increased relative kidney weights without kidney lesion in males of the 200 mg/kg group were considered to be treatment related. Pelvic dilatation in female and solitary cyst in the medullar and subcapsular solitary cyst in males were noted in the kidney in the 200 mg/kg groups, and they were single occurrences and observed spontaneously.

No abnormalities were noted in the FOB, body weights and urinalysis.

In the recovery groups, mottled teeth were observed in males of the 200 mg/kg group, This was considered related to dysplasia of the enamel. In an animal which has brown teeth like a rat, decreased iron pigments in the ameloblast, impaired iron-pigment secretion to the enamel and degeneration or necrosis of the enamel are generally observed by fluoride ingestion (Dokusci Byori Soshiki Gaku, 2000). In this study, these signs were not observed in histopathological examinations at the end of the dosing and recovery periods. Therefore, the mottled teeth were considered to be slight and reversible changes, and considered disappeared with the growing of teeth because of no histopathological changes. Decreased MCV in males, decreased eosinophils, decreased total cholesterol, increased absolute and relative kidney weights and increased relative ovary weights were considered to be no treatment-related since they were slight changes within the background values in our laboratory. No abnormalities were noted in body weights, food consumption, FOB, urinalysis and histopathological examinations.

Applicant's summary and conclusion