Potassium 1,1,2,2,3,3,4,4,4-nonafluorobutane-1-sulphonate

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2001

Report date:

2001

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Potassium 1,1,2,2,3,3,4,4,4-nonafluorobutane-1-sulphonate

EC Number:

249-616-3

EC Name:

Potassium 1,1,2,2,3,3,4,4,4-nonafluorobutane-1-sulphonate

Cas Number:

29420-49-3

Molecular formula:

C4HF9O3S.K

IUPAC Name:

potassium 1,1,2,2,3,3,4,4,4-nonafluorobutane-1-sulfonate

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Lot 2
- Expiration date of the lot/batch: No data
- Purity test date: 17 April, 2000

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature, protected from light.
- Stability under test conditions: Dosing solutions were considered stable withing 14 days of preparation. All formulated test materials were stored refrigerated in amber glass bottles.
- Solubility and stability of the test substance in the solvent/vehicle: The test article was dissolved in carboxymethylcellulose. Samples were retained predose and on the last day of dosing for each batch and were analysed for proper formulation.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test article was dissolved in carboxymethylcellulose.
- Final preparation of a solid: The test article was dissolved in carboxymethylcellulose to allow dosing via oral gavage at a dose volume of 25 mL/kg.

FORM AS APPLIED IN THE TEST: The test article was dissolved in carboxymethylcellulose.

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories,Inc.
- Females (if applicable) nulliparous and non-pregnant: Yes
- Age at study initiation: Approximately 8 weeks old.
- Weight at study initiation: Males: 209 to 253 grams, Females: 178 to 212 grams
- Fasting period before study: No data
- Housing: Individually in screen-bottom stainless steel cages.
- Diet (e.g. ad libitum): Teklad Certified Rodent Diet (#8728) ad libitum
- Water (e.g. ad libitum): Tap water ad libitum
- Acclimation period: 10 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 28-26
- Humidity (%): 30-70
- Air changes (per hr): No data
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 31 August, 2000 To: 13 October, 2000

Administration / exposure

Route of administration:

oral: gavage

Details on route of administration:

The doses were administered at 25 mL/kg body weight.

Vehicle:

CMC (carboxymethyl cellulose)

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS: The test article was dissolved in 1% carboxymethylcellulose in water.

VEHICLE
- Concentration in vehicle: Dosing solutionswere formulated to allow for dosing of 0 (vehicle control), 100, 300, and 900 mg/kg at a dose volume of 25 mL/kg body weight.
- Amount of vehicle (if gavage): 25 mL/kg body weight
- Lot no.: 69H0028

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Dose concentrations were verified using HPLC/MS

Duration of treatment / exposure:

Daily for 28 days.

Frequency of treatment:

Daily

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0 mg/kg bw (total dose)

Remarks:

Vehicle control (1% CMC in water).

Doses / concentrations 2

Dose / conc.:

100 mg/kg bw (total dose)

Doses / concentrations 3

Dose / conc.:

300 mg/kg bw (total dose)

Doses / concentrations 4

Dose / conc.:

900 mg/kg bw (total dose)

No. of animals per sex per dose:

10 (5 additional/sex in the control and high dose groups as recovery animals).

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Based on a 10-day range-finding study.
- Rationale for animal assignment: Random
- 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: Observations for mortalityand moribundity were recorded twice daily (a.m. and p.m.). Cageside observations were recorded at approximately one to two hours postdose daily and when a change was noted.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Detailed clinical observations were recorded once before the first dose and at least weekly there after or when a change was noted.

BODY WEIGHT: Yes
- Time schedule for examinations: Body weights were recorded pretest,on Study Day 1, approximately weekly, prior to terminal fasting, and prior to necropsy (fasted weight). The pretest body weights are not included in this report but are located in the raw data. The fasted body weights recorded prior to necropsy were used only for the calculation of relative organ weights.

FOOD EFFICIENCY:
- Feed consumption was measured approximately weekly.

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Blood samples for hematological analysis were obtained via cardiac puncture prior to necropsy. The animals were anesthetized with carbon dioxide for the collection. Feed was withheld overnight prior to collection.
- Anaesthetic used for blood collection: No, blood collected prior to necropsy.
- Animals fasted: Yes
- How many animals: All
- Parameters checked: The blood samples were evaluated for the following parameters: total and relative differential leukocyte counts, erythrocyte counts, hemoglobin concentration, hematocrit value, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, platelet counts, prothrombin time, and activated partial thromboplastin time

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Blood samples for hematological analysis were obtained via cardiac puncture prior to necropsy. The animals were anesthetized with carbon dioxide for the collection. Feed was withheld overnight prior to collection.
- Anaesthetic used for blood collection: No, blood collected prior to necropsy.
- Animals fasted: Yes
- How many animals: All
- Parameters checked: The samples were processed and evaluated for the following parameters: total protein, albumin, globulin, albumin/globulin ratio, glucose, cholesterol, triglycerides, total bilirubin, urea nitrogen, creatinine, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, gamma-glutamyltransferase, calcium, phosphorus, sodium, potassium, and chloride.

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: DuringWeek 4, each animal was assessed for sensory reactivity to stimuli of different types (e.g., auditory and visual), grip strength, and motor activity
- Dose groups that were examined: All
- Battery of functions tested: sensory activity, grip strength, motor activity

IMMUNOLOGY: No

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
Organ weights were recorded for the following organs prior to fixation: adrenal glands, brain, heart, kidneys, liver, ovaries, spleen, thymus, and testes. Paired organs were weighed together. The following organs and tissues were examined in situ, dissected free, and fixed in 10% neutral buffered formalin: adrenal glands, aorta, bone marrow (sternum), brain (brain stem, cerebellum, cerebrum), cervix, epididymides, esophagus, eyes (with optic nerve), femur (with articular surface), gross lesions, Harderian gland, heart, large intestine (cecum, colon, rectum), small intestine (duodenum, ileum, jejunum), kidneys, lacrimal gland, liver, lungs (with bronchi), lymph nodes (mesenteric, mandibular), mammary gland, ovaries, oviducts, pancreas, pituitary gland, prostate gland, salivary gland (mandibular), sciatic nerve, seminal vesicles, skeletal muscle, spinal cord (cervical, thoracic, lumbar), spleen, stomach (forestomach, glandular), testes, thymus, thyroid/parathyroid, tongue, trachea, uterus, urinary bladder, vagina, and Zymbal's gland.

HISTOPATHOLOGY: Yes
Histopathology was performed on all tissues from teh vehicle control, high-dose and recovery animals and on all gross lesions.

The following tissues were examined: adrenal glands, aorta, bone marrow (sternum), brain (brain stem, cerebellum, cerebrum), cervix, epididymides, esophagus, eyes (with optic nerve), femur (with articular surface), gross lesions, Harderian gland, heart, large intestine (cecum, colon, rectum), small intestine (duodenum, ileum, jejunum), kidneys, lacrimal gland, liver, lungs (with bronchi), lymph nodes (mesenteric, mandibular), mammary gland, ovaries, oviducts, pancreas, pituitary gland, prostate gland, salivary gland (mandibular), sciatic nerve, seminal vesicles, skeletal muscle, spinal cord (cervical, thoracic, lumbar), spleen, stomach (forestomach, glandular), testes, thymus, thyroid/parathyroid, tongue, trachea, uterus, urinary bladder, vagina, and Zymbal's gland.

Statistics:

Means and standard deviations were calculated for all quantitative data. Treated groups of the same sex were compared to Group 1 (vehicle control) at common time points using an ANOVA with a post hoc Dunnett's t Test (Dunnett, C.W., J. Amer. Statis. Assoc., 50:1096-1121, 1955) or another appropriate test. A two-sided c_of 0.05 or 0.01 was used to determine if statistically significant differences existed between the vehicle control and the treated groups.

Results and discussion

Results of examinations

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

Clinical observations consisted of localized alopecia (back, limbs, neck, and underside), urine stained abdominal fur, scabbed area on back and neck, and soft or liquid feces. These findings are considered incidental and unrelated to testmaterial administration.Additional cage side observations were all normal.

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

There were no treatment-related effects on body weights for either Main Study or Recovery animals. On StudyDay 36, the mean bodyweight in males that received 900 mg/kg/day test article was decreased by 7% when compared to the corresponding controls. This finding was considered incidental. Throughout the study, bodyweight changes intreatment groups were calculated and compared to the corresponding control groups with no significant findings recorded for either Main Study or Recovery animals

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

Throughout the study, absolute feed consumption per treatment group was calculated and compared to the corresponding control groups with no significant findings recorded for either Main Study or Recovery animals.

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

not examined

Haematological findings:

effects observed, non-treatment-related

Description (incidence and severity):

The reported significant hematological findings are considered incidental and not associated with test material administration.

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

Main Study findings in males that received _>300mg/kg/day T-7485 indicate significant decreases in serum phosphorus (approximately 16% of the control values) and potassium (approximately 20% of the control values). In contrast, an increase in chloride levels in males that received 900 mg/kg/day test article (4% above the control values) was recorded. In the absence of similar findings in Recovery animals, the changes in phosphorus, potassium, and chloride levels appear to be associated with test material administration but do not appear to be toxicologically or biologically significant, as indicated by historical data. Other incidental findings do not appear to be related to administration of the test material.

Urinalysis findings:

not examined

Behaviour (functional findings):

effects observed, non-treatment-related

Description (incidence and severity):

Assessment of neurological effects on animals treated with the test article included peripheral neuropathy, motoractivity, and audio/visual evaluations. There were no test material-related effects on motoractivity or audio/visual.

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

Absolute and relative male liver (between 25% to 30%) and female kidney (9% to 11%) weights were significantly increased in animals that received 900 mg/kg/day test article when compared to the corresponding controls. These significant changes were not recorded during the recovery period, and since there does not appear to, be an effect of body weight on organ development, these data suggest the changes are test material-related. However, the biological and toxicological significance is unclear due to the lack of any supportive gross or histopathological findings. The remaining significant findings are considered incidental and unrelated to test material administration.

Gross pathological findings:

effects observed, non-treatment-related

Description (incidence and severity):

Only one gross lesion was recorded at necropsy and was not considered to be test material-related.

Neuropathological findings:

effects observed, non-treatment-related

Description (incidence and severity):

Assessment of neurological effects on animals treated with the test article included peripheral neuropathy, motoractivity, and audio/visual evaluations. There were no test material-related effects on motoractivity or audio/visual.

Histopathological findings: non-neoplastic:

effects observed, non-treatment-related

Description (incidence and severity):

The microscopic lesions observed in the test animals are considered to be incidental and in no way related to receiving the test material.

Histopathological findings: neoplastic:

no effects observed

Effect levels

Key result

Dose descriptor:

NOAEL

Effect level:

ca. 900 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

behaviour (functional findings)

body weight and weight gain

clinical biochemistry

clinical signs

gross pathology

haematology

histopathology: neoplastic

histopathology: non-neoplastic

mortality

organ weights and organ / body weight ratios

Target system / organ toxicity

Key result

Critical effects observed:

no

Applicant's summary and conclusion

Conclusions:

Based on the results of the study, the No Observed Adverse Effect Level (NOAEL) is 900 mg/kg/day.

Executive summary:

The subacute oral toxicity of the test article was evaluated in Sprague Dawley rats following 28 consecutive daily doses. This study was performed in compliance with OECD GLP (1981). The study design was based on OECD 407. The test material was prepared in 1% carboxymethyl-cellulose in deionized water (vehicle). Rats (10/sex/dose) received 0 (vehicle), 100, 300, or 900 mg/kg/day of the test article via oral gavage at a dose volume of 25 mL/kg for 28 consecutive days. In addition, recovery groups (5/sex/dose) received 0 or 900 mg/kg/day of the test article in a similar manner. With the exception of the recovery groups, all groups were euthanized 24 hours following the last dose. The recovery groups were euthanized following a 14-day recovery period. Parameters evaluated: clinical observations (pretest and at least weekly thereafter or when a change was noted), body weights (weekly), food consumption (weekly), neurological function observations (Week 4), gross necropsy (termination), hematology (termination), clinical chemistry (termination), organ weights of select organs (termination), and histopathology of select organs and tissues (termination). All animals survived. Localized alopecia and scabbed area on back and neck were observed among all dose groups. At 900 mg/kg, one male exhibited urine-stained abdominal fur, and another male exhibited soft or liquid feces. These clinical observations were considered incidental and unrelated to test material administration. There were no test article-related effects on body weights or food consumption. Absolute and relative male liver (25% to 30%) and female kidney (9% to 11%) weights were significantly increased in animals that received 900 mg/kg/day test article when compared to the corresponding controls. These significant changes were not recorded during the recovery period, and since there does not appear to, be an effect of body weight on organ development, these data suggest the changes are test material-related. However, the biological and toxicological significance is unclear due to the lack of any supportive gross or histopathological findings. The remaining significant findings are considered incidental and unrelated to test material administration. No toxicologically relevant changes were noted in the clinical chemistry and hematology parameters evaluated or tissues subjected to histopathological evaluation. Based on the results of the study, the No Observed Adverse Effect Level (NOAEL) is 900 mg/kg/day.