Hexafluoropropene, oxidized, oligomers, reduced, fluorinated

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:

July 1992 to August 1992

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

some missing details and examinations

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1993

Report date:

1993

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

(Paris, 1981)

Deviations:

no

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Hexafluoropropene, oxidized, oligomers, reduced, fluorinated

EC Number:

500-537-5

EC Name:

Hexafluoropropene, oxidized, oligomers, reduced, fluorinated

Cas Number:

161075-00-9

Molecular formula:

R-O(C3F6O)m-R with R= - CF3, - C2F5, -CF2H

IUPAC Name:

1,1,1,2,3,3-hexafluoro-2,3-bis(1,1,2,2,2-pentafluoroethoxy)propane; 1,1,1,2,3,3-hexafluoro-2-(1,1,2,2,2-pentafluoroethoxy)-3-(trifluoromethoxy)propane; 1,1,1,2,3,3-hexafluoro-3-(1,1,2,2,2-pentafluoroethoxy)-2-(trifluoromethoxy)propane; 1,1,1,2,3,3-hexafluoro-3-{[1,1,1,2,3,3-hexafluoro-3-(trifluoromethoxy)propan-2-yl]oxy}-2-(trifluoromethoxy)propane; 1,1,1,3,3,4,6,6,7,9,9,10,12,12,12-pentadecafluoro-4,7,10-tris(trifluoromethyl)-2,5,8,11-tetraoxadodecane; 1-(difluoromethoxy)-1,1,2,3,3,3-hexafluoro-2-(1,1,2,2,2-pentafluoroethoxy)propane; 2,2,3,5,5,6-hexafluoro-3,6-bis(trifluoromethyl)-1,4-dioxane; 2-(difluoromethoxy)-1,1,1,2,3,3-hexafluoro-3-(1,1,2,2,2-pentafluoroethoxy)propane

Details on test material:

- Name of test material (as cited in study report): GALDEN HT 70
- Substance type: pure substance
- Physical state: liquid (density: 1.69 g/ml)
- Analytical purity: 100%
- Impurities (identity and concentrations): none known
- Composition of test material, percentage of components: not reported
- Isomers composition: not reported
- Purity test date: not reported
- Lot/batch No.: G 30
- Expiration date of the lot/batch: December 1996
- Stability under test conditions: not reported
- Storage condition of test material: room temperature

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: livestock farming
- Age at study initiation: males: 30 days, females: 25 days
- Weight at study initiation: males: 75-85 g, females: 60-70 g
- Fasting period before study: not reported
- Housing: 2 or 3 animals/sex/cage. Wire cages with stainless steel feeder. Cage size(cm):40.5 x 38.5 x 18h
- Diet (e.g. ad libitum): Certificated pelleted diet, supplied with vitamins and trace elements. Available ad libitum.
- Water (e.g. ad libitum): from municipal water main system, filtered and distributed ad libitum.
- Acclimation period: two weeks, the rats were housed in a quarantine room an their health status was assessed by daily clinical observations.

ENVIRONMENTAL CONDITIONS
- Temperature: 22°C +- 2°C
- Humidity: 55 % +- 10%
- Air changes (per hr): approximately 20 air changes/hour
- Photoperiod (hrs dark / hrs light): 12 hour cicle (7 a.m.-7 p.m.)

IN-LIFE DATES:
Males: From: 8-Jul-1992 To 5-Aug-1992
Females: From 9-Jul-1992 To 6-Aug-1992

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: 0.5 (w/v) Methylcellulose 400 cps water solution additioned with 0.25% (w/v) Polysorbate 80 (Tween 80)

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:
Every day appropriate amounts of substance were weighed and emulsified with vehicle in suitable containers in order to obtain the final concentration of 25, 50, 100 mg/ml to be administered, respectively, to the animals of tested groups at constant administration volume of 10 ml/kg b.w. . The suspension were kept magnetically stirred until the end of the daily administration.
Formulates were prepared just prior to the administration.

VEHICLE
- Justification for use and choice of vehicle (if other than water): not reported
- Concentration in vehicle: 25, 50 100 mg/ml at the doses of 250, 500 and 1000 mg/kg/day, respectively.
- Amount of vehicle (if gavage): 10 ml/kg b.w.
- Lot/batch no. (if required): Methylcellulose 400 cps: Batch No.300535, Polysorbate 80 (Tween 80) Batch No.7245049
- Purity: not reported

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Samples for concentration check were collected from formulates prepared on July 8, 1992 (week 1) and July 30, 1992 (week 4) and stored at -20 °C until dispatch to the Sponsor for analysis.

Duration of treatment / exposure:

28 days, no recovery.

Frequency of treatment:

once a day, for 7 days a week, for the entire study period.

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0 mg/kg bw/day (nominal)

Remarks:

vehicle control group

Doses / concentrations 2

Dose / conc.:

250 mg/kg bw/day (nominal)

Doses / concentrations 3

Dose / conc.:

500 mg/kg bw/day (nominal)

Doses / concentrations 4

Dose / conc.:

1 000 mg/kg bw/day (nominal)

No. of animals per sex per dose:

5

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: not reported; highest dose is the highest recommended dose.
- Rationale for animal assignment: Randomization.
The 40 (20M + 20F) rats were selected from a larger goup (27M + 27F) than the one required: before commencement of treatment all the animals were weighed: animals at extreme of the body weight range or showing eye abnormalities at the pre-trial examination were discarded.
The required number of rats (20M + 20F) was allocated to the dosage groups by means of a computerized stratified sequenced randomization program.

Positive control:

Not included

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily Intervals.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Daily Intervals. Physical appearance, bahaviour and clinical signs.

BODY WEIGHT: Yes
- Time schedule for examinations: Weekly Intervals. Each animal was weighed prior to the beginning of the treatment period (day 0, the day before the first administration) and then at weekly intervals throughout the study period.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Not applicable
Food consumption was recorded, for each cage, at weekly intervals throughout the study period. Food comsuption was calculated as the difference between the known offered amount per cage and the remainder recorded after 7 days. Individual food intake was then calculated, in g/animal/day, and reported in tables, for each 7-day period.

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No
Water consumption was not measured
Compound intake calculation: not applicable

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: Prior to the start of treatment (males: day -1, females: day -2) and before the end of the dosing period (males: day 27, females day 27)
- Dose groups that were examined: All animals
Eyes were examined with a direct ophtalmoscope, after instillation of 0.5% Tropicamide.

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at the end of dosing period, day 29.
- Anaesthetic used for blood collection: Yes (ether)
- Animals fasted: Yes
- How many animals: All animals
- The following parameters were examined: Erythrocytes, Hemoglobin, Leukocytes, Neutrophils, Lymphocytes, Monocytes, Eosinophils, Basophils, Hematocrit, Mean corpuscolar volume, Mean corpuscolar HGB conc, Mean corpuscolar HGB, Platelets, Prothrombin time.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at the end of dosing period, day 29.
- Animals fasted: Yes
- How many animals: All animals
- The following parameters were examined: Glucose, Urea, Creatinine, Total bilirubin, Alkaline phosphatase, SGOT, SGPT, Total protein, Albumin, Globulin, A/G ratio, Alpha 1 globulins, Alpha 2 globulins, Beta globulins, Gamma globulins, Total cholesterol, Sodium, Potassium Calcium, Chloride, Inorganic phosphorus.

URINALYSIS: Yes
- Time schedule for collection of urine: at the end of dosing period, day 29.
- Metabolism cages used for collection of urine: Yes.
- Animals fasted: Yes (16 hours)
In order to collect urine samples, on the day before the scheduled analysis the animals received 10 ml/kg of tap water (by gavage) as water load; subsequently they were kept in metabolism cages for about 16 hours without food and water.
- The following parameters were examined: Diuresis, Specific gravity, pH, Protein, Bilirubin, Glucose, Ketones, Blood, Leukocytes, Urobilinogen, Nitrites, Epithelial cells, Leukocytes, Erythrocytes, Phosphate crystals, Urate crystals, Calcium oxalate crystals, Casts, Bacteria.

NEUROBEHAVIOURAL EXAMINATION: No (not part of the test guideline at the time of the study)

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
At the end of the treatment period the body weight of each animal that had been fasted overnight was recorded before the animal was killed .

The following organs were removed:
skin and mammary gland, urinary bladder, prostate, testes, epididymides, seminal vesicles, vagina, uterus, ovaries, spleen, stomach, duodenum, jejunum, ileum, cecum, colon, rectum, mesenteric lymph nodes, pancreas, liver, kidneys, adrenals, submandibular salivary glands and lymph nodes, sternum with bone marrow, heart, thymus, lungs, aorta, trachea, esophagus, thyroid with parathyroid if present in the thyroid section, eyes, Harder's lacrimal glands, tongue, brain: coronal section at threee levels, pituitary, skeletal muscle: biceps femoris, peripheral nerve: sciatic nerve, spinal cord: thoracic, cervical and lumbar, vertebrae, femur including articular surface, gross lesions.

The following organs were removed, trigger and weighed:
testes, ovaries, spleen, liver, kidneys, adrenals, heart, brain
Individual organ weight/fasted body weight ratio was calculated.

HISTOPATHOLOGY: Yes
Histology was carried out on the following organs:
Spleen, stomach, duodenum, ileum, colon, liver, kidneys, adrenals, heart, and gross lesions found at macroscopic examination.

Statistics:

The parameters statistically examined were:
- body weight
- body weight gain
- food intake
- hematology
- blood chemistry
- urinalysis (except the semi-quantitative analysis and the microscopic examination of the sediment of the urine)
- organ weights (absolute and relative to body weight)

The above-reported data were compared and tested according a decisiontree which includes Bartlett's test, ANOVA, Dunnett's multiple test and Mann Withney's test.
Level of significance was reported for all determinations.

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Description (incidence and severity):

No clinical changes were seen at any dose in either males or females.

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Description (incidence and severity):

No body weight changes of toxicological relevance were seen in either males or females at any dose and no variations from the control values that attained statistical relevance were recorded at any time.
A very slight retarded growth was however observed in animals treated at the highest dosage (1000 mg/kg/day). In this group, when compared to the basal body weight, a mean body weight gain slightly lower than controls was observed in males starting from week 1 onward with evidence of a time relationship (from 3% at week 1 to 8% at week 4); the same trend, even though confined to the final weighing (week 4) was seen in females for which the differential value of about 5% was mainly related to the low body weight gain of 2 out of the 5 dosed rats.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

No effects on the quantity of food consumed that could be related to treatment were seen in either males and femals; the mean food intake data were similar in all the experimental groups and there were no individual values outside the norm.

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

no effects observed

Description (incidence and severity):

No eye changes were seen in any animal.

Haematological findings:

effects observed, non-treatment-related

Description (incidence and severity):

No changes in haematologic profile of toxicological relevance were seen at any dose in either males and females at the investigation performed at the end of the dosing period. The statistical analysis did not reveal any significant difference from the control values in females treated at any dose or in male treated at 250 and 500 mg/kg/day.
In males given 1000 mg/kg/day the mean 1000 RBC number, hemoglobin, and hematocrit values were slightly higher than in controls; the changes significant at the statistical analysis, involved most of the animals of this group (Incidence: 80% ) with values above the range of the controls, but again within the normal range of variability. Moreover, since the there were no changes in the erythrocyte indices, these slight modifications were considered of no toxicological relevance.

Clinical biochemistry findings:

no effects observed

Description (incidence and severity):

No blood chemistry changes that could be related to GALDEN LMW administration were seen at any dose in either males and females.
The statistical analysis highlighted a few significant differences from the control values in animals treated at 250 and 1000 mg/kg/day: in males sodium and chloride ion levels were slightly higher at both doses while in females a glucose level higher (at 250 mg/kg/day) or total birilubin and SGOT values lower (at 1000 mg/kg/day) were observed. Since these modifications were very slight and without any individual value outside the normal range of variability they were considered incidental in origin.

Urinalysis findings:

no effects observed

Description (incidence and severity):

No compound-related changes were seen.
For the parameters statistically analysed (diuresis, specific gravity and pH), no significant differences from the control value were seen in either males or females treated at any dose.
Assessment of the semiquantitative analysis and sediment microscopy did not reveal any notable differences from controls in any group; the positive reactions for proteins in the urine of 3 out of the 5 male rats treated at 500 mg/kg/day were considered incidental since the findings was always slight in degree (grade 1), confined to this dose only, and without other concurent urinary parameter modifications.

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

None of the noted organ weight variations were regarded as related to the compound treatment. In fact the range of these changes were included within the accumulated control backgroud data in the laboratoty.
In particular, the absolute and relative adrenal weight increase observed in all male treated groups is regarded as of no relation to treatment. In fact the individual values were included within range of individual adrenal weight seen in control rats of other experiments performed in the laboratory.
In addition no treatment related histological alteration was noted in this organ. The reduction in absolute testicular weight observed in the high dosage group was caused by an incidental reduction in tested weight of a single rat. This testicular reduction in weight was sporadically noted in control rats of the same age used in other experiments in the laboratory.

Gross pathological findings:

no effects observed

Description (incidence and severity):

There were a range of lesions observed in control and treated groups, none of them was regarded as related to test article treatment.
In fact, the incidences were comparable among treated and control rats and/or they were characteristically observed in untreated rats of the same age used as controls in other experiments carried out in the laboratory.
Examples of noted lesions included: hepatic accentuation of lobular pattern, presence of whitish areas and congestion in various organs.

Histopathological findings: non-neoplastic:

effects observed, non-treatment-related

Description (incidence and severity):

None of the observed lesions is regarded as related to the compound administration.
All observed lesions had comparable incidence in control and treated groups and/or they are characteristically noted in untreated rats of the same age used in other experiments carried out in the laboratory. Among the most frequently observed lesions were hepatocytic mononuclear cell aggregation (incidence: 30%), hepatocyte vacuolation (incidence: 23%) and renal tubular basophilia (incidence: 6%).
In particular one female rat treated with the high dose had a single focus of degeneration with inflammation in the heart (myocard). This lesion is sporadically noted in control rats of the same age. Due to its rare occurrence in the present study, it is concluded to be of no relation to the compound administration.
Moreover, congestion or hemorrhage and edema seen in the cecum of group 4 female no. 5661 and in the prostate of goup 1 male no. 5630 were considered incidentally induced at autopsy by needle trauma during the intraperitoneal injection of barbiturate to induce anesthesia.
Abdominal cavity subacute inflammation seen in group 4 female no 5664, consistent at gross pathology examination with a hernia, is an event that can sporadically occur also in untreated rats.

Effect levels

Key result

Dose descriptor:

NOEL

Effect level:

>= 1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no effects observed at the limit dose

Remarks on result:

not determinable due to absence of adverse toxic effects

Target system / organ toxicity

Key result

Critical effects observed:

no

Applicant's summary and conclusion

Conclusions:

No compound-related deaths, clinical signs or abnormalities were observed. Body weight showed only a very slight retarded growth at the highest dose which was considered of no toxicological relevance in absence of other effects. Food intake was unaffected by treatment.
Laboratory investigations and post-mortem examinations did not reveal changes attributable to GALDEN LMW administration.
In conclusion, GALDEN LMW when daily administered at the dosage of 250, 500 and 1000 mg/kg/day to Sprague Dawley by oral route for 4 weeks, was well tolerated up to and including the highest dosage administered.

Executive summary:

In this study, conducted under GLP according to OECD method No. 407, the effects of repeated administration of the test article GALDEN LMW on Sprague Dawley rats were evaluated.

GALDEN LMW, emulsified in 0.5% (w/v) Methylcellulose 400 cps water solution, additioned with 0.25% (w/v) Tween 80, was administered by oral route (gavage) once a day for 4 consecutive weeks to groups of Sprague Dawley Crl:CD rats at the doses of 250, 500 and 1000 mg/kg/day.

The test article formulates were kept under magnetic stirring during administration.

The administration volume was kept constant at 10 mg/kg bw in all treated groups, the test article concentration in the vehicle being varied accordingly (25, 50 and 100 mg/ml, respectively). The volume administered was adjusted on the basis of most recently recorded body weight (individual values).

Control animals received 10 ml/kg/day of 0.5 % (w/v) Methylcellulose 400 cps water solution, additioned with 0.25% (w/v) Tween 80.

Each experimental group consisted of 5 males and 5 females.

At the end of the 4-week dosing period, all animals were killed for pathology studies.

 

No animals died during the study.

No clinical signs were seen in any animal.

No body weight changes of toxicological relevance were seen at any dose. Moreover, at the highest dose, a minor body weight variation consistent with a slightly retarded growth was observed in males from week 1 onward and in females at the final (week 4) measurement.

Food consumption was unaffected by GALDEN LMW oral administration at all the dose levels applied.

No eye abnormalities were observed at ophthalmoscopic examination.

No changes of toxicological relevance were seen in the hematological profile of either males and females at any dose.

No blood chemistry or urinary changes that could be related to treatment were seen at any dose.

No changes deemed of compound-related origin were seen at post-mortem examination.

 

In conclusion GALDEN LMW, when daily administered at the dosage of 250, 500 and 1000 mg/kg/day to Sprague Dawley rats by oral route for 4 weeks, was well tolerated up to and including the highest dosage administered.