1-Propene, 1,3,3,3-tetrafluoro-

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian cell study: DNA damage and/or repair

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Reference

Endpoint:

short-term repeated dose toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 412 (Subacute Inhalation Toxicity: 28-Day Study)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Route of administration:

inhalation: gas

Type of inhalation exposure:

nose only

Vehicle:

other: unchanged (no vehicle)

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

total carbon analysis with flame ionization detector

Duration of treatment / exposure:

6 hours day

Frequency of treatment:

5 days a week

Dose / conc.:

5 000 ppm (analytical)

Remarks:

Group 2: Low dose

Dose / conc.:

20 000 ppm (nominal)

Remarks:

Group 3: Mid dose

Dose / conc.:

50 000 ppm (analytical)

Remarks:

Group 4: High dose

No. of animals per sex per dose:

5

Control animals:

yes, sham-exposed

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

DETAILED CLINICAL OBSERVATIONS: No

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION:
- Food consumption for per group determined and mean daily diet consumption

WATER CONSUMPTION: No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: ay scheduled necropsy
- Anaesthetic used for blood collection: Yes (identity) -Nembutal
- Animals fasted: Yes
- How many animals: all survivors
- Parameters listed in guideline were examined.


CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at scheduled necropsy
- Animals fasted: Yes
- How many animals: all survivors
- Parameters listed in guideline were examined.

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No


OTHER:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily


DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily


BODY WEIGHT: Yes / No / No data
- day s -1, -1, 0, 7 and 14


FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes


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: Yes / No / No data


HAEMATOLOGY: Yes
- Time schedule for collection of blood: at sacrifice
- Anaesthetic used for blood collection: Yes (Nembutol)
- Animals fasted: Yes
- How many animals: all
- Parameters checked in table were examined.


CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood:
- Animals fasted: Yes
- How many animals:all
- Parameters checked in table were examined.

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes- as listed in guideline plus full respiratory tract and nasal passages

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

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):

Red blood cells and thrombocytes increased and the mean corpuscular haemoglobin concentration decreased in males of the hlgh
concentration group and the prothrombin time was increased in female animals of the mid- and high concentration groups. Monocyte count was increased in the male animals of the high concentration group.

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

Glucose, alanine aminotransferase, aspartate aminotransferase, albumin and urea were increased and the levels of cholesterol and phospholipids and sodium were decreased in male animals of the high-concentration group. The levels of aspartate aminotransferase and urea were also increased in the male animals of the midconcentration group.

In female animals of the high concentration group, the levels of alkaline phosphatase, afanine aminorransferase and urea were increased and cholesterol was decreased. In the mid concentration group the level of cholesterol was also decreased and the level of aspartate aminotransferase was
increased.

Endocrine findings:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

In the high concentration group absolute and relative lung weights were decreased in males only. Absolute and relative liver
weights were increased in both male and female animals of the high-concentration
group

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

Exposure-related pale appearance of the livers of almost all animals of the high-concentration group. The two other findings,
a flabby kidney in a control female and an abdominal cyst in a hlgh concentration male are common findings in n rats of this strain and age.

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Microscopic examination revealed exposure-related histopathological changes in the heart of the mid- and high-concentration males and females, in the liver of the mid- and high-concentration males and high-concentration females, and, in the nasal passages of the high-concentration males and females. Mononuclear cell infiltrates in the heart were most prominent in the animals of the mid-concentration group. This histopathological change is, therefore, an example of a non-monotonic concentration related increase in the response to an exposure. Because the mononuclear cell infiltrates in the heart of the mid- and high-concentration groups were considered to be related to the exposure, the mononuclear cell infiltrates in the heart seen in at least one female animal of the low concentration group cannot be excluded to be related to the exposure as well.

Liver - more pronounced histopathological changes in exposed males compared to exposed females were in accordance with the changes in lives related clinical chemistry parameters.

Nasal passages - goblet cell expression was locally decreased in the nasal passages of high-concentration males and females and this was also found in a few mid- and low-concentration males and females. In the latter groups there was, however, no clear concentration-response relationship in incidence and degree and, therefore, it was concluded that decreased goblet cell expression in the intermediate concentration animals was not related to exposure.

Histopathological findings: neoplastic:

no effects observed

Other effects:

not examined

Dose descriptor:

NOAEC

Effect level:

5 000 ppm

Sex:

male/female

Basis for effect level:

other: Hematologic analysis, clinical chemistry analysis, organ weight measurements, and macroscopic and microscopic examination of the heart, liver and nasal passages revealed treatment-related effects in animals exposed to 20,000 and 50,000 ppm.

Critical effects observed:

not specified

Conclusions:

The main effects of exposure of rats to 5000, 20000 and 50000 ppm HFO-1234ze for 6 hours a day, 5 days a week consisted of concentration related effects in heart and liver in animals of the mid and high concentration groups and in the nasal passages of animals of the high concentration group.

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Reference

Endpoint:

short-term repeated dose toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

OECD Guideline 412 (Subacute Inhalation Toxicity: 28-Day Study)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Route of administration:

inhalation: gas

Type of inhalation exposure:

nose only

Vehicle:

other: unchanged (no vehicle)

Details on inhalation exposure:

same as decribed for the 13 week study

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

same as described for the 13 week study

Duration of treatment / exposure:

6 hours day

Frequency of treatment:

5 days a week

Dose / conc.:

1 000 ppm (nominal)

Remarks:

Group 2: Low dose

Dose / conc.:

5 000 ppm (nominal)

Remarks:

Group 3: Low-mid dose

Dose / conc.:

10 000 ppm (nominal)

Remarks:

Group 4: High-mid dose

Dose / conc.:

15 000 ppm (nominal)

Remarks:

Group 5: High dose

No. of animals per sex per dose:

5 females and 5 males per dose

Control animals:

yes, sham-exposed

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

DETAILED CLINICAL OBSERVATIONS: No

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION:
- Food consumption for per group determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes

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: Yes

WATER CONSUMPTION: No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at scheduled necropsy
- Anaesthetic used for blood collection: Yes (identity) -Nembutal
- Animals fasted: Yes
- How many animals: all survivors
- Parameters checked as listed in guideline


CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at scheduled necropsy
- Animals fasted: Yes
- How many animals: all survivors
- Parameters checked in table [No.?] were examined.

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

Sacrifice and pathology:

GROSS PATHOLOGY: Yes as listed in OECD guideline
HISTOPATHOLOGY: Yes as listed in OECD guideline with additional respiratory tract tissues

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

no effects observed

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

not specified

Ophthalmological findings:

not specified

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Endocrine findings:

not examined

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Very slight to moderate inflammation of the heart found only in the male rats in the 15000 ppm exposure level group.

Histopathological findings: neoplastic:

no effects observed

Other effects:

not examined

Dose descriptor:

NOAEL

Effect level:

10 000 ppm

Sex:

male/female

Basis for effect level:

other: very slight to moderate inflammation of the heart found only in the male rats in the 15000 ppm exposure level group

Critical effects observed:

not specified

Conclusions:

In this study, groups of rats were exposed to levels of 0 (control), 1000, 5000, 10000 or 15000 ppm of HFO-1234ze for 6-hrs/day, 5 days/wk for 4 weeks. In addition to the normal toxicology assessment, two genetic toxicity assessments were conducted, Unscheduled DNA synthesis (UDS) and Bone marrow micronucleus formation. No genetic effects were observed. Slight to moderate inflammation was observed in the hearts of males exposed to 15000 ppm. The no-observed adverse effect concentration was determined to be 10000 ppm.

Executive summary:

In this study, groups of rats were exposed to levels of 0 (control), 1000, 5,000, 10,000 or 15,000 ppm of HFO-1234ze for 6-hrs/day, 5 days/wk for 4 weeks. In addition to the normal toxicology assessment, two genetic toxicity assessments were conducted, Unscheduled DNA synthesis (UDS) and Bone marrow micronucleus formation. No genetic effects were observed. Slight to moderate inflammation was observed in the hearts of males exposed to 15000ppm. The no-observed adverse effect concentration was determined to be 10000 ppm.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2006

Report date:

2006

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 486 (Unscheduled DNA Synthesis (UDS) Test with Mammalian Liver Cells in vivo)

GLP compliance:

yes (incl. QA statement)

Type of assay:

unscheduled DNA synthesis

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

471-480-0

EC Name:

-

Cas Number:

1645-83-6

Molecular formula:

Hill formula: C3H2F4 CAS formula: C3H2F4

IUPAC Name:

(1E)-1,3,3,3-tetrafluoroprop-1-ene

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male

Administration / exposure

Route of administration:

inhalation: gas

Duration of treatment / exposure:

28 days

Frequency of treatment:

6 hours/d, 5 days/wk for 4 weeks

Post exposure period:

none

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

5 000 ppm (analytical)

Remarks:

Group 2: Low dose.

Doses / concentrations 2

Dose / conc.:

15 000 ppm (nominal)

Remarks:

Group 3: High dose.

No. of animals per sex per dose:

5

Control animals:

yes, sham-exposed

Positive control(s):

2-acetylaminofluorene
- Justification for choice of positive control(s): sustance known to produce unscheduled DNA synthesis
- Route of administration: oral gavage
- Doses / concentrations: 50 mg/kg
- Dose volume 10 ml/kg of a 5 ml/kg solution

Examinations

Tissues and cell types examined:

cultured hepatocytes

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION:


TREATMENT AND SAMPLING TIMES ( in addition to information in specific fields): Isolated hepatocytes were prepared within 24 hours of the last expsosure for negative control and test article treated animals. Hepatocytes from AAF treated animals were prepared 12-16 hours after exposure

Evaluation criteria:

The study is considered valid if the positive control gives a positive response and if the negative control gives a clear negative response.
A response at a data point is considered positive if the population average NNG > 5, and if at least 20 % of the cells are "in repair" and weakly positive if the population average NNG is between 0 and 5.
A test material is considered to cause DNA damage and induce DNA repair in liver cells if at least one dose level at one time point results in an increase in the mean NNG compared to the vehicle control.

A test material is considered non-genotoxic under the conditions of the test if all dose levels and time points produce NNG < O.
Both numerical significance and biological relevance are considered together in the evaluation.

Results and discussion

Test results

Key result

Sex:

male

Genotoxicity:

negative

Toxicity:

no effects

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

other: did not meet criteria but were different from controls and HFO-1234ze treated

Additional information on results:

Both the test material HFO-1234yf and the negative control (clean air) yielded net nuclear grains (NNG)< O. Since exposure to the test material did not induce NNG> 5, it is demonstrated that HFO-1234ze did not induce unscheduled DNA synthesis in rat hepatocytes. The positive control substance 2-AAF induced NNG3.25 with less 20 % of the cells in repair. While the positive control did not exhibit as robust examination as expected, the results did show that these animals had a higher NNG and % cells in repair compared to controls or HFO-1234ze tested cells.

Any other information on results incl. tables

The mean NNG determined for 2 -AAF (-6.310 was clearly higher than the NNG of the negative control (-9.78) or test substance (-13.04 and -11.82). The cells in repair was 3.25% for 2 -AAF and <0.5 for control and 0.2% for the test substance. No technical problems were noted and the negative controls and background counts were within historical controls. The response of the groups treated with HFO-1234ze is considered to be correct.

Applicant's summary and conclusion

Conclusions:

It is concluded that the test material HFO-1234ze did not induce unscheduled DNA synthesis (UDS) in liver cells of male rats, exposed to the test material at concentrations up to 15000 ppm by inhalation, under the conditions used in this study.