Registration Dossier

Administrative data

Description of key information

Several repeated dose toxicity studies are available (2-weeks, 4-weeks and 13-weeks of exposure). No toxicity was observed in male or female rats exposed to concentrations up to 5 000 ppm for 6 hours/day for 2, or 13 weeks, or concentrations up to 10 000 ppm following 4 weeks of exposure.   

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records
Reference
Endpoint:
sub-chronic toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: OECD guideline study conducted under GLP
Reference:
Composition 0
Qualifier:
according to
Guideline:
OECD Guideline 413 (Subchronic Inhalation Toxicity: 90-Day Study)
GLP compliance:
yes (incl. certificate)
Limit test:
no
Test material information:
Composition 1
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland
- Age at study initiation: 7-8 weeks
- Weight at study initiation: mean weights 252 g (males) and 176 g (female)
- Fasting period before study: none
- Housing: macrolon cages with wood shaving beding
- Diet (e.g. ): ad libitum (overnight fast prior to necropsy)
- Water (e.g. ):ad libitum
- Acclimation period: at least 7 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): 36-46 (one day humidity in high dose group was > 70% for less than 5 minutes- quickly fixed)
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12


IN-LIFE DATES: From: March 2007 To: June 2007
Route of administration:
inhalation: gas
Type of inhalation exposure:
nose only
Vehicle:
other: unchanged (no vehicle)
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus:cylindrical PVC column with a volume of ~ 70 liters surrounded by a transparent hook. The test atmosphere was introduced at the bottom and exhausted at the top
- Method of holding animals in test chamber:
- Source and rate of air: at least 1 lter/min
- Temperature, humidity in air chamber: 20-24C; 30-70%humidity
- Air flow rate: at least 1 liter/min


TEST ATMOSPHERE
- Brief description of analytical method used: total carbon analysis
- Samples taken from breathing zone: yes


Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
total carbon analysis
Duration of treatment / exposure:
6 hours day
Frequency of treatment:
5 days a week
Remarks:
Doses / Concentrations:
0, 1500, 5000 and 15000 ppm
Basis:
analytical conc.
No. of animals per sex per dose:
10
Control animals:
yes, sham-exposed
Details on study design:
- Dose selection rationale: 15000 ppm (1.5%) was chosen due to the effects observed in the range finding study
- Post-exposure recovery period in satellite groups: none
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

DETAILED CLINICAL OBSERVATIONS: Yes (daily)

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

OPHTHALMOSCOPIC EXAMINATION: yes

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 listed in OECD 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 OECD guideline were examined.

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: no
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes - those organs listed in the guideline plus nose (6-levels), larynx (3 levels), trachea (3 levles including bifurcation), and each lung lobe at 1 level.
Statistics:
Data were evaluated by the appropriate statistical test (one-way analysis of variance followed by Dunnett's multiple comparison test, one-way analyis of variance (ANOVA) followed by Dunn't multiole comparison testes, Krisckal-Wallis nonparametric Anova followed by Mann-Whitney U-tests, Fischers exact probability test.
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):
no effects observed
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:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Histopathological findings: neoplastic:
no effects observed
Details on results:
HAEMATOLOGY - concentration of the monocytes and thrombocytes were increased in male animals of the high concentration group.


CLINICAL CHEMISTRY - increased AST in high dose males, increased urea in high dose females


HISTOPATHOLOGY: NON-NEOPLASTIC-multifocal mononuclear cell infiltrates, often accompanied by myocardial degeneration (increased eosinophilia and pyknotic nuclei). A silver stain for reticulum did not provide evidence for fibrosis in high dose male and female

Dose descriptor:
NOEC
Effect level:
5 000 ppm (analytical)
Sex:
male/female
Basis for effect level:
other: multifocal mononuclear cell infiltrates in the heart of the 15000ppm males and females.
Critical effects observed:
not specified
Conclusions:
Overall, exposure of rats to 1500, 5000 or 15,000 ppm HFO-1234ze for 6 h/day, 5 days/week for 63 or 64 exposure days over a 91-92 day period did result in slight adverse effects in animals of the high concentration group only. In the present subchronic inhalation toxicity study, the mid concentration level of 5000 ppm was therefore considered to be a No-Observed-Adverse-Effect-Level (NOAEL) for male and female rats.
Executive summary:

Overall, exposure of rats to 1500, 5000 or 15,000 ppm HFO-1234ze for 6 h/day, 5 days/week for 63 or 64 exposure days over a 91-92 day period did result in slight adverse effects in animals of the high concentration group only. In the present subchronic inhalation toxicity study, the mid concentration level of 5000 ppm was therefore considered to be a No-Observed-Adverse-Effect-Level (NOAEL) for male and female rats.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEC
23 300 mg/m³
Study duration:
subchronic
Species:
rat
Quality of whole database:
There are 3 repeated dose toxicity studies available (2-week, 4-week and 13-week exposure duration). All studies were performed recently and according to relevant Guidelines. All studies are considered bo be of good quality (K1).

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Mode of Action Analysis / Human Relevance Framework

Additional information

In the 2 -week study, rats were exposed to 0, 5000, 20000 and 50000 ppm. The NOEC in that study was 5000 ppm. In the 4 -week study, rates were exposed to 0, 1000, 5000, 10000 and 15000 ppm leading to a NOEC of 10000 ppm. Finally in the 13 -week study, rats were exposed to 0, 1500, 5000 and 15000 ppm leading to a NOEC of 5000 ppm.

The multifocal mononuclear cell infiltrates in the heart of both sexes in the 13 -week study was only seen at 15000 ppm and was predominately characterized as slight. Similar effects were observed in the 2 - and 4 - week studies. Increased exposure duration however did not appear to increase the incidence or severity of the effects seen in the heart. This statement was confirmed by the observations made in the 2 -generation reproductive toxicity study. Additionally, fibrosis was not observed in the 90 day (13 -week) study indicating this is not a progressive lesion. Based on these results, 15 000 ppm was considered as the LOAEC and 5 000 ppm was considered the NOEC for the 13 -week exposure study. This NOEC is used further on in the dossier for risk assessment purposes. However, when considering the lack of progression of the heart lesion, the NOEC of 10 000 ppm seen in the 4 -week study could represent a more accurate NOEC for repeat exposures to HFO-1234ze.


Repeated dose toxicity: inhalation - systemic effects (target organ) cardiovascular / hematological: heart

Justification for classification or non-classification

Repeated inhalation exposure to HFO-1234ze resulted in cardiac effects (multifocal infiltrates) which did not increase in incidence or severity with prolonged exposure. The no observed effect level in the 13 -week study is 5000 ppm and for the 4 - week study 10 000 ppm. Based on these results, classification is not justified.