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Toxicological information

Genetic toxicity: in vivo

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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-referenceopen allclose all
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.
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

Constituent 1
Chemical structure
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 / concentrationsopen allclose all
Dose / conc.:
5 000 ppm (analytical)
Remarks:
Group 2: Low dose.
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.