Registration Dossier

Administrative data

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2004
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study was conducted in compliance with OECD GLP (1997) regulations with the excetion of a final report being written.

Data source

Reference
Reference Type:
other company data
Title:
Unnamed
Year:
2004

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents)
Deviations:
yes
Remarks:
The study was completed but a final report was never written.
GLP compliance:
yes
Remarks:
Study was conducted in compliance with GLP regulations but a final report was not written.
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: Liquid
Details on test material:
- Name of test material (as cited in study report): T-7867
- Substance type: mono-constituent
- Physical state: Liquid
- Analytical purity: 98.5%
- Lot/batch No.: Lot 4 with spikes
- Expiration date of the lot/batch: 30 December 2005
- Storage condition of test material: Refridgerated
- Other:

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: Approximately 6 weeks
- Weight at study initiation: Male mean: 207 g, Female mean: 160.5 g
- Fasting period before study: No data
- Housing: 5 animals per sex per cage in Macrolon plastic cages (type IV, height 18 cm).
- Diet (e.g. ad libitum): Altromin (code VRF 1) pelleted lab diet, ad libitum
- Water (e.g. ad libitum): Tap water, ad libitum
- Acclimation period: At least 5 days before the start of treatment

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-24 C
- Humidity (%): 30-70
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 16 January 2004 To: 26 February 2004

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: Perfluorohexane: PF-5060 (FC-72)
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:

VEHICLE
- Justification for use and choice of vehicle (if other than water): Solubility and to reduce test article volatility.
- Concentration in vehicle: The test article was dosed at 50, 150, and 450 mg/kg/day.
- Amount of vehicle (if gavage): No data
- Lot/batch no. (if required): 040183
- Purity: No data
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The analytical method was based on NOTOX project 393497.
Duration of treatment / exposure:
Animals were dosed once daily for 28 days.
Frequency of treatment:
Daily for 28 days.
Doses / concentrations
Remarks:
Doses / Concentrations:
0, 50, 150, 450 mg/kg/day
Basis:
actual ingested
No. of animals per sex per dose:
5/sex/dose at 50 and 150 mg/kg/day and 10/sex/dose at 0 and 450 mg/kg/day at allow for recovery groups.
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale:
- Rationale for animal assignment (if not random): Random
- Rationale for selecting satellite groups: Random for the control and high (450 mg/kg/day) groups.
- 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: At least twice daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: At least once daily.

BODY WEIGHT: Yes
- Time schedule for examinations: Treatment period: Days 1, 8, 15, 22, and 28. Recovery period: Days 1, 8, and 14.

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: Both eyes of each animal were examined on week 4 (all groups) and week 6 (recovery groups- only if ophthalmoscopic changes are seen in week 4).
- Dose groups that were examined: All

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Immediately prior to scheduled necropsy (Day 28 for treatment groups and Day 14 of recovery period for recovery groups).
- Anaesthetic used for blood collection: Yes (iso-flurane).
- Animals fasted: Yes, overnight
- How many animals: All
- Parameters checked: Erythocyte count, hemoglobin, hematocrit, mean corpuscular colume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, platelet count, red cell distribution width, total leucocyte count, differential leucocyte count.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Immediately prior to scheduled necropsy (Day 28 for treatment groups and Day 14 of recovery period for recovery groups).
- Animals fasted: Yes, overnight
- How many animals: All
- Parameters checked: Alanine aminotransferase, alkaline phosphatase, aspartate aminotransferase, gamma-glutamyl transferase, bilirubin, chloride, cholesterol, creatinine, glucose, phosphorus, protein, albumin protein, globulin, albumin globulin ratio, urea, triglycerides, calcium, potassium, sodium.

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: On week 4 of treatment
- Dose groups that were examined: All animals
- Battery of functions tested: Hearing ability, pupillary reflex, static righting reflex and grip strength.
Sacrifice and pathology:
GROSS PATHOLOGY: Yes, all animals surviving to the end of the observation period and all moribund animals were anesthetized using iso-flurane and subjected to gross necropsy. Descriptions of all macroscopic abnormalities were recorded.

HISTOPATHOLOGY: Yes. The following tissues were collected and examined: adrenal glands, aorta, caecum, cervix, clitoral gland, colon, duodenum, epididymides, eyes with optic nerve and harderian gland, female mammary gland area, femur (including joint), heart, ileium, jejunum, kidneys, larynx, lacriminal gland, liver, lung, lymph nodes, nasopharynx, esophagus, ovaries, pancreas, peyer's patches, pituitary gland, prostate gland, rectum, salivary glands, sciatic nerve, seminal vesicles, skeletal muscle, skin, spinal cord, spleen, sternum with bone marrow, stomach, testes, thymus, thyroid, tongue, trachea, urinary bladder, uterus, vagina, any gross lesion.

Results and discussion

Results of examinations

Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
no effects observed
Haematological findings:
effects observed, treatment-related
Clinical biochemistry findings:
effects observed, treatment-related
Urinalysis findings:
not examined
Behaviour (functional findings):
no effects observed
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
not examined
Details on results:
CLINICAL SIGNS AND MORTALITY: All animals survived to scheduled necropsy. No adverse clinical signs were noted in any animals in any dose group

BODY WEIGHT AND WEIGHT GAIN: Females in the 450 mg/kg group had statically significantly higher mean body weights compared to controls at the end of treatment. Body weights were not statistically significantly different between the control 450 mg/kg groups at the end of the recovery period in males or females.

OPHTHALMOSCOPIC EXAMINATION: No adverse ophthalmolic effects were noted upon examination.

HAEMATOLOGY: Upon hematological analysis, males had statistically significantly lower RBC (450 mg/kg group), HGB (150 and 450 mg/kg groups), HCT (450 mg/kg group), MCH (50, 150, 450 mg/kg groups), MCHC (50, 150, 450 mg/kg groups), and PT (450 mg/kg group) and statically significantly higher RDW (450 mg/kg group) compared to controls. At the end of the recovery period MCHC, and RDW were still statistically significantly elevated in the 450 mg/kg group compared to controls. Upon hematological analysis, females had statistically significantly lower RBC (450 mg/kg group), HGB (50 and 450 mg/kg groups), HCT (450 mg/kg group), MCV (450 mg/kg group), MCHC (50 and 450 mg/kg groups), and statically significantly higher RDW (150 and 450 mg/kg group) compared to controls. At the end of the recovery period HGB was still statistically significantly higher than controls in the 450 mg/kg group.

CLINICAL CHEMISTRY: Males had statistically significantly elevated bilirubin (150, 450 mg/kg groups) and potassium (450 mg/kg group) compared to controls. At the end of the recovery period, ASAT was statistically significantly decreased in 450 mg/kg-treated males and total protein was elevated compared to controls. Females had statistically significantly elevated ALAT (450 mg/kg group), GGT (450 mg/kg group), cholesterol (150, 450 mg/kg groups), potassium (450 mg/kg group), calcium (450 mg/kg group), chloride (450 mg/kg group) compared to controls. At the end of the recovery period, ALAT and calcium were statistically significantly decreased and creatinine was increased compared to controls in females.

NEUROBEHAVIOUR: No animals had abnormal observations in the functional battery tests.

ORGAN WEIGHTS: At the end of treatment, males had statically significantly decreased brain and testes weights compared to controls that had lost statistical significance by the end of the recovery period compared to controls. At the end of treatment, females had statistically significantly increased liver weight and spleen weight that had lost statistical significance by the end of recovery compared to controls.

GROSS PATHOLOGY: No abnormal finidings were observed in any animals upon gross necropsy.

HISTOPATHOLOGY: NON-NEOPLASTIC: No abnormal findings were observed in any animals upon microscopic examination.

Effect levels

Dose descriptor:
NOAEL
Effect level:
ca. 450 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
other: overall effects clinical signs; mortality; body weight; ophthalmoscopic examination; haematology; clinical chemistry; gross pathology; organ weights; histopatholog

Target system / organ toxicity

Critical effects observed:
not specified

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 450 mg/kg-day.
Executive summary:

The subacute oral toxicity of the test article (clear, colorless liquid; CASRN 84650-68-0, purity 98.5%; Lot 4, with spikes) was evaluated in Wistar Rats. METHODS: This study was performed under OECD GLP (1997) conditions. The study method was based on OECD 407 (1995), MHLW No. 1039 and METI No. 1014 (1986), U.S. EPA OPPTS 870.3050 (2000), and EC Directive 96/54/EC, Annex IV.D (1996). Rats (5/sex/dose) received 0, 50, 150, or 450 mg/kg-day test article via oral gavage for 28 days followed by a 14-day recovery period. The test article was diluted in perfluorohexane (PF-5060, FC-72) for dosing. Body weights and clinical and functional observations were recorded. Clinical hematological and biochemical parameters were measured. Microscopic examinations were performed on all animals from the control and 450 mg/kg-day dose groups and on animals of all dose groups that died spontaneously or were sacrificed in extremis. All animals survived to scheduled necropsy. No adverse clinical signs were noted in any animals in any dose group. Females in the 450 mg/kg group had statically significantly higher mean body weights compared to controls at the end of treatment. Body weights were not statistically significantly different between the control 450 mg/kg groups at the end of the recovery period in males or females. Upon hematological analysis, males had statistically significantly lower RBC (450 mg/kg group), HGB (150 and 450 mg/kg groups), HCT (450 mg/kg group), MCH (50, 150, 450 mg/kg groups), MCHC (50, 150, 450 mg/kg groups), and PT (450 mg/kg group) and statically significantly higher RDW (450 mg/kg group) compared to controls. At the end of the recovery period MCHC, and RDW were still statistically significantly elevated in the 450 mg/kg group compared to controls. Upon hematological analysis, females had statistically significantly lower RBC (450 mg/kg group), HGB (50 and 450 mg/kg groups), HCT (450 mg/kg group), MCV (450 mg/kg group), MCHC (50 and 450 mg/kg groups), and statically significantly higher RDW (150 and 450 mg/kg group) compared to controls. At the end of the recovery period HGB was still statistically significantly higher than controls in the 450 mg/kg group. Males had statistically significantly elevated bilirubin (150, 450 mg/kg groups) and potassium (450 mg/kg group) compared to controls. At the end of the recovery period, ASAT was statistically significantly decreased in 450 mg/kg-treated males and total protein was elevated compared to controls. Females had statistically significantly elevated ALAT (450 mg/kg group), GGT (450 mg/kg group), cholesterol (150, 450 mg/kg groups), potassium (450 mg/kg group), calcium (450 mg/kg group), chloride (450 mg/kg group) compared to controls. At the end of the recovery period, ALAT and calcium were statistically significantly decreased and creatinine was increased compared to controls in females. At the end of treatment, males had statically significantly decreased brain and testes weights compared to controls that had lost statistical significance by the end of the recovery period compared to controls. At the end of treatment, females had statistically significantly increased liver weight and spleen weight that had lost statistical significance by the end of recovery compared to controls. No abnormal findings were noted upon gross necropsy of all animals or histopathological examination of the control and 450 mg/kg-day groups. Based on the results of the study, the No Observed Adverse Effect Level (NOAEL) of the test article is 450 mg/kg-day.