1,1,1,2,2,3,4,5,5,5-decafluoro-3-methoxy-4-(trifluoromethyl)pentane

Repeated dose toxicity: oral

Administrative data

Endpoint:

repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2004

Report date:

2004

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

Version / remarks:

1995

Deviations:

no

Remarks:

No deviations ocurred that negatively impacted the integrity of the study.

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Novec 7300

IUPAC Name:

Novec 7300

Details on test material:

- Name of test material (as cited in study report): Novec 7300
- Physical state: Colorless, clear liquid
- Analytical purity: 99.85%

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 3M Company, Lot 1
- Expiration date of the lot/batch: No data
- Purity test date: No data

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature in the dark.
- Stability under test conditions: The stability of the test substance during the dosing period was confirmed via IR spectrum before and after dosing.
- Solubility and stability of the test substance in the solvent/vehicle: Stable

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test article was suspended in olive oil.

Test animals

Species:

rat

Strain:

Sprague-Dawley

Details on species / strain selection:

Sprague Dawley rats are considered an appropriate strain for a repeated dose study.

Sex:

male/female

Administration / exposure

Route of administration:

oral: unspecified

Vehicle:

olive oil

Details on oral exposure:

Method of administration:
oral gavage

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

In the homogeneity test. top, middle and bottom layers of 10.0 and 0.10 w/v% formulations \vere taken (n=3) just after preparation. The samples were mixed wilh ethyl acetate (5,000 rimes concentrated), and quantitatively analyzed by gas chromatography (GC). The homogeneity of formulations was confllllled with a relative standard deviation in each layer. In the stability test, the homogeneity samples were stored at the cold and dark place. Middle layer of the sample was taken (n=3) after 3 days and 7 days. These samples were diluted with ethyl acetate (5,000 x concentrated), and the samples were quantitatively analyzed by GC to confirm the stabi lity as compared to the concentrations immediately after preparation (results of the homogeneity sludy).

Duration of treatment / exposure:

Test duration: 28 days

Frequency of treatment:

Dosing regime: 7 days/week

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0 mg/kg bw/day (nominal)

Remarks:

Control (12/sex)

Doses / concentrations 2

Dose / conc.:

25 mg/kg bw/day (nominal)

Remarks:

6/sex

Doses / concentrations 3

Dose / conc.:

150 mg/kg bw/day (nominal)

Remarks:

6/sex

Doses / concentrations 4

Dose / conc.:

1 000 mg/kg bw/day (nominal)

Remarks:

12/sex

No. of animals per sex per dose:

Male: 12 animals at 0 mg/kg bw/day
Male: 6 animals at 25 mg/kg bw/day
Male: 6 animals at 150 mg/kg bw/day
Male: 12 animals at 1000 mg/kg bw/day
Female: 12 animals at 0 mg/kg bw/day
Female: 6 animals at 25 mg/kg bw/day
Female: 6 animals at 150 mg/kg bw/day
Female: 12 animals at 1000 mg/kg bw/day

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Based on a range-finder
- Rationale for animal assignment (if not random): Random
- Rationale for selecting satellite groups: Control and high-dose groups per guideline
- Post-exposure recovery period in satellite groups: 14 days

Positive control:

NA

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: During the dosing period, the general condition was observed 3 times a day, i.e., before dosing (in the morning), during and after dosing, and in the afternoon, daily from day 1 to day 28. During the recovery period, the general condition was observed once a day in the morning daily.

BODY WEIGHT: Yes
- Time schedule for examinations: Body weight was measured on day - 2 (at allocation to groups) before initiation of dosing, on days 1, 3, 8, 12, 17, 21,26 and 28 during the dosing period, and on days 1, 5, 10 and 14 (recovery) during the recovery period. In addition, immediately before necropsy, body weights were measured for calculation of relative organ weights.

FOOD CONSUMPTION: Food consumption was measured once before initiation of dosing, on days 3, 8, 15, 22 and 28 during the dosing period, and on days 4, 8 and 14 (recovery) during the recovery period.

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Blood or plasma samples obtained by blood sampling from the abdominal aorta under ether anesthesia after overnight fasting (16 to 20 hr) at completion of the dosing period (excluding the recovery groups) and at completion of the recovery period were determined for the following items.
- Anaesthetic used for blood collection: Yes
- Animals fasted: Yes
- How many animals: All
- Parameters examined:
Red blood cell count (RBC)
White blood cell count (WBC)
Hemoglobin conc. (Hb) (g/dL)
Hematocrit value (Ht) (%)
Mean corpuscular volume (M.CV) (fL)
Mean corpuscular hemoglobin (MCH) (pg)
Mean corpuscular hemoglobin conc. (MCHC) (g/dL)
Platelet count (Platelet)
Reticulocyte count
Prothrombin time (PT) (sec)
Activated partial thromboplastin time (APTT) (sec)
Differentiation of leukocytes


CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Blood or plasma samples obtained by blood sampling from the abdominal aorta under ether anesthesia after overnight fasting (16 to 20 hr) at completion of the dosing period (excluding the recovery groups) and at completion of the recovery period were determined for the following items.
- Animals fasted: Yes
- How many animals: All
- Parameters evaluated:

Aspartate aminotransferase (AST)
Alanine aminotransferase (AL T) (IU/L)
Alkaline phosphatase (ALP) (TU/L)
Cholinesterase (ChE) (TU/L)
y-Giutamyl transpeptidase (y-GTP) (IU/L)
Total cholesterol (T-Cho)
Triglyceride (TG)
Glucose
Total protein (T-Protein)
Albumin
AIG ratio
Blood urea nitrogen (BUN)
Creatinine
Total bilimbin (T-Bil)
Calcium (Ca)
Inorganic phosphorus (IP)
Sodium (Na)
Potassium (K)
Chloride (C1)

URINALYSIS: Yes
- Time schedule for collection of urine: Urinalysis was performed once (day 28) during the dosing period (excluding the recovery groups) and once (day 14 (recovery)) during the recovery period. Urine samples (accumulated for 15-17 hr) collected in individual metabolic cages ( ISO Wx200 Dx263 H mm) were detennined with drinking water ad lib.
- Metabolism cages used for collection of urine: Yes
- Animals fasted: No
- Parameters evaluated:
Urine volume
Color
Turbidity
Osmolarity
pH
Protein
Glucose
Occult blood
Urinary sediments

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: Before dosing and once weekly thereafer
- Dose groups that were examined: All animals
- Battery of functions tested: sensory activity / grip strength / motor activity /

IMMUNOLOGY: No

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
The weights of the following organs were measured in all animals. The relative organ weight was also calcu lated based on the body weight at the time of necropsy.
Liver, heart, kidneys, testes, epid idymides, ovaries, brain, spleen, thymus and adrenals

HISTOPATHOLOGY: Yes

Respiratory system: trachea, lungs (filled with fixative)
Digestive system: incisors, stomach, intestine (duodenum to rectum, with Peyer's patches), liver
Cardiovascular system: Heart
Urinary system: Kidneys, urinary bladder
Reproductive system: testes, epididymides, prostate, seminal vesicle, ovaries, uterus, vagina
Nervous system: brain (cerebrum, cerebellwn and pon), spinal cord, sciatic nerve
Hematopoietic and lymphatic: bone marrow (femur), axillar and mesenteric lymph nodes, spleen, thymus
Endocrine system: Pituitary gland, thyroids, adrenals
Special sense organ: Eyeball

Light microscopic examinations were performed for the following organs and tissues after embedding in paraffin, sectioning and hematoxylin and eosin staining. The histological preparation except for incisors, testes and epididymides was performed in Sapporo General Pathology Laboratory according to the assignment protocol.

trachea, lungs, incisors, forestomach, glandular stomach, duodenum, jejunum, ileum, cecum, colon, rectum, liver, heart, kidneys, urinary bladder, testes, epididymides, prostate, seminal vesicles, ovaries, uterus, vagina, cerebrum, cerebellum, prostate, spinal cord, sciatic nerve, bone marrow, axillary and mesenteric lymph nodes, spleen, thymus, pituitary gland, thyroids, parathyroids, adrenals, eyeball

Results and discussion

Results of examinations

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

Mottled teeth were observed in males and females of the 1000 mg/kg groups from day 26 onwards.

Mortality:

no mortality observed

Description (incidence):

No treatment-related mortality was observed during the study.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

No treatment-related body weight changes were identified during the study.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

No treatment-related food consumption changes were noted during the study.

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

Concerning the hematology, APTT was increased in males of the ISO and 1,000 mg/kg groups and in the females of the 1,000 mg/kg group.

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

Concerning the blood chemistry, cholinesterase and albumin were increased in males of the 150 and 1,000 mg/kg groups. A/G ratio was increased in males and females of the 1,000 mg/kg groups. Increased ALT, ALP, y-GTP and BUN and decreased chloride were noted in males of the 1,000 mg/kg group. Increased albumin and decreased creatinine were noted in females of the 1,000 mg/kg group. These changes were considered to be related to the liver and kidney effects of the test substance.

In the recovery test, increased ALT was noted in males of the 1,000 mglkg group. A/G ratio was increased in females of the 1,000 mglkg group. They were considered to be reversible since the severity and frequency were decreased in comparison with those at end of the dosing period.

Urinalysis findings:

effects observed, treatment-related

Description (incidence and severity):

Concerning the urinalysis, increased urine volume, decreased uosm, light colored urine and increased red blood cells, white blood cells and epithelial cells were noted in males of the 1,000 mg/kg group.

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

Concerning the organ weights, relative liver weights were increased in males and females of the 150 and 1,000 mg/kg groups. Absolute Liver weights were increased in males of the 150 and 1,000 mg/kg groups and in females of the 1,000 mg/kg group. Absolute and relative kidney weights and relative spleen weight were increased in males of the 1,000 mg/kg group.

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

In the necropsy, enlargement of the liver was noted in males of the 150 and 1,000 mg/kg groups and in females of the 1,000 mg/kg group. Mottled teeth, rough surface of the incisors, whitish region in the liver and enlargement of the kidneys were noted in males of the 1,000 mg/kg group.

The recovery groups (1,000 mg/kg) did not show liver enlargement.

Neuropathological findings:

no effects observed

Description (incidence and severity):

No treatment-related changes were noted in the functional observation battery results.

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

In the liver, centrilobular hypertrophy of the hepatocytes was noted in males (150 and 1000 mg/kg) and females (1000 mg/kg). Focal necrosis of the hepatocytes was noted in males (1000 mg/kg). In the kidney, basophilic tubules with the prominent nuclei, dilatation of the tubules and necrosis and hyperplasia of the collecting tubular epithelium were noted in males of the 1000 mg/kg group.

In the incisor, decreased pigments in the ameloblasts and hypomineralization of the dentin were noted in males and females of the 1000 mg/kg group. Atrophy, degeneration and irregular alignment of the ameloblasts, inhibition of the matrix withdrawal and irregular alignment of the dentinal tubules were noted in males (1000 mg/kg). In the pituitary gland and thyroid, hypertrophy of the thyrotrophs in the pituitary gland and follicular cell hypertrophy in the thyroid were noted in males (1000 mg/kg). In the spleen, congestion was notes in males (1000 mg/kg).

In the recovery group, increased absolute and realtive kidney weights and basophilic tubuels with prominent nucleoli and dialation of the tubules in the kidney were noted in males of the 1000 mg/kg/day group. The severity and frequency of the findings were decreased compared to those at the end of the dosing period.

Histopathological findings: neoplastic:

no effects observed

Description (incidence and severity):

No neoplastic lesions were noted during the study.

Description (incidence and severity):

Based on the recovery group, effects on the incisor, liver, kidney, spleen, pituitary gland and thyroid were considered to be reversible.

Effect levels

Key result

Dose descriptor:

NOAEL

Effect level:

150 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

histopathology: non-neoplastic

Target system / organ toxicity

1

Key result

Critical effects observed:

yes

Lowest effective dose / conc.:

1 000 mg/kg bw/day (nominal)

System:

urinary

Organ:

kidney

Treatment related:

yes

Dose response relationship:

yes

2

Key result

Critical effects observed:

yes

Lowest effective dose / conc.:

1 000 mg/kg bw/day (nominal)

System:

hepatobiliary

Organ:

liver

Treatment related:

yes

Dose response relationship:

yes

Applicant's summary and conclusion

Conclusions:

Based on the results of the study, the No Observed Adverse Effect Level (NOAEL) of the test article is 150 mg/kg/day.

Executive summary:

The subacute oral toxicity of T-7869 was evaluated inSprague Dawley rats following 28 consecutive days of dosing. This study was performed in compliance with OECD GLP (1997) and Japanese MHLW, METI, and MOE GLP guidelines (2003). The study design was based on Japanese testing guidelines (2003), OECD 407 (1995), and Council DirectiveB.7 (1996). T-7869 was prepared in olive oil (vehicle) once weekly. Rats (6/sex/group) received 0, 25, 150, or 1000 mg/kg T-7869 via oral gavage at dosevolume of 10 mL/kg once daily for 28 consecutive days. One vehicle control group and one 1000 mg/kg/day dose group served as recovery groups and wereobserved for 14 days following the last dose. All dose groups, except for the recovery groups, were euthanized after the last dose. The recovery groups were euthanized after the 14 day observation period. Parameters measured: clinical observations (at least once daily), body weight (twice weekly), food intake(approximately weekly), functional observation battery (week 4), gross necropsy (termination), clinical chemistry (termination), urinalysis (termination), andweight and histopathology of select organs (termination). All animals survived. During the dosing period, mottled teeth (15/24) and salivation (18/24)were observed in the 1000 mg/kg dose group. Salivation, loss of hair, scab formation, and/or exudate were observed in the 25 and 150 mg/kg dose groups.Salivation and mottled teeth were also observed among vehicle control group animals. During the recovery period, mottled teeth (12/12) and partial loss oflower incisors (3/12) were observed in the 1000 mg/kg dose group. There were no abnormal changes in body weight or food intake.  No changes in organ weight and/or histopathology of the thymus and lymph nodes. No toxicologically relevant changes in immunological parameters. Enlargement of the kidney was noted in the males at 1000 mg/kg. In the kidney, basophilic tubules with the prominent nuclei, dilation of the tubules and necrosis and hyperplasia of the collecting tubular epithelium were noted in males of the 1000 mg/kg group. Effects were considered reversible. Absolute and relative liver weights were increased in males in the 150 and 1000 mg/kg groups. Relative liver weights were increased in females in the 150 and 1000 mg/kg groups. Absolute liver weights were increased in females in the 1000 mg/kg group. Enlargement of the liver was observed in males at 150 and1000 mg/kg and in females at 1000 mg/kg. In the liver, centrilobular hypertrophy of the hepatocytes was noted in males of the 150 and 1000 mg/kg groups and in the females of the 1000 mg/kg group. Focal necrosis of the hepatocytes was noted in males of the 1000 mg/kg group. At termination of the dosing period, total bilirubin was decreased in males and females of the 150 and 1000 mg/kg groups. Increased albumin was noted in 1000 mg/kg-treated females and in 150 and 1000 mg/kg-treated males. AST was decreased in 1000 mg/kg-treated females and in 25 mg/kg-treated males and females. In addition, 1000 mg/kg-treated males had increased ALT, alkaline phosphatase, A/G ratio, and BUN. The females in the 1000 mg/kg group had increased A/G ratio and decreased creatinine. At the termination of the recovery period, ALT was increased in 1000 mg/kg-treated males, and A/G ratio was increased in 1000 mg/kg-treated females. Based on the results of the study, the No Observed Adverse Effect Level (NOAEL) of the test article is 150 mg/kg/day.