Registration Dossier

Administrative data

Description of key information

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
Study period:
2015-04-22 to 2016-05-06
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study
Qualifier:
according to
Guideline:
OECD Guideline 413 (Subchronic Inhalation Toxicity: 90-Day Study)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.29 (Sub-Chronic Inhalation Toxicity:90-Day Study)
Deviations:
no
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Crj: CD(SD)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Inc., Raleigh, NC
- Age at study initiation: 38 days old at receipt, 7 weeks old at the initiation of test substance exposures
- Weight at study initiation: Individual body weights ranged from 236 g to 283 g for males and from 172 g to 214 g for females at the initiation of test substance exposures
- Housing: 2 or 3 per cage
- Diet : PMI Nutrition International, LLC, Certified Rodent LabDiet® 5002 (meal), ad libitum, except during the exposure periods and except during the period of fasting prior to necropsy
- Water : Reverse osmosis-treated (on-site) drinking water, delivered by an automatic watering system. Ad libitum except during the exposure periods.
- Acclimation period: 14 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Actual mean daily temperature ranged from 19.0°C to 22.7°C during the study
- Humidity (%): actual mean daily relative humidity ranged from 43.0% to 55.8% during the study
- Air changes (per hr): minimum of 10 fresh air changes per hour
- Photoperiod : 12 hour light (0600 hours to 1800 hours)/12 hour dark

IN-LIFE DATES: From: 07May2015 To: 06Aug2015
Route of administration:
inhalation: gas
Type of inhalation exposure:
whole body
Vehicle:
air
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: 4 approximately 1000-L stainless steel and glass whole body inhalation exposure chambers
- Method of holding animals in test chamber: whole body inhalation exposure chambers
- source of air: filtered air
- Method of conditioning air: A HEPA-filter and an activated-charcoal bed were used to pre-treat room air prior to delivery to chambers.
- Temperature, humidity, pressure in air chamber: mean temperature 19 to 25°C, mean humidity 30 to 70%, under slight negative pressure.
- Air flow rate: 200 to 250 Standard liters per minute
- Air change rate: 12 to 15 ACH
- Treatment of exhaust air: Chamber exhaust was passed through the facility exhaust system consisting of redundant exhaust blowers preceded by activated-charcoal and HEPA-filter units.

TEST ATMOSPHERE
- Brief description of analytical method used: Analyzed exposure concentrations were determined at approximately 45-minute intervals using an appropriate gas chromatography (GC) method
- Samples taken from breathing zone: yes

CONTROL GROUP
Concurrent exposure to humidified, filtered air on a comparable regimen
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The test substance usage was documented by weighing the gas cylinder prior and following exposure.
Analyzed exposure concentrations were determined at approximately 45-minute intervals using an appropriate gas chromatography (GC) method after calibration with standards prepared at 5 gas concentrations. The first sampling round initiated approximately 18 min after the start of exposure atmosphere generation.
- Homogeneity was assessed during method development: 4 remote locations.

Duration of treatment / exposure:
6 hours per day on a 5-day per week basis for 13 weeks (65 total exposures)
Frequency of treatment:
5-day per week
Remarks:
Doses / Concentrations:
0, 80, 280, and 1000 ppm
Basis:
nominal conc.
Remarks:
Doses / Concentrations:
0, 81, 279, and 1004 ppm
Basis:
analytical conc.
No. of animals per sex per dose:
10 animals
Control animals:
other: humidified, filtered air on a comparable regimen
Details on study design:
- Dose selection rationale: based on results of an OECD 422 study
Positive control:
No
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily for mortality and moribundity

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: detailed physical examinations were performed weekly

BODY WEIGHT: Yes
- Time schedule for examinations: Individual body weights were recorded twice weekly (± 1 day) during the first 4 weeks of the exposure period, then weekly for the remainder of the study.

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
Food consumption was recorded once weekly (± 2 days) beginning after randomization

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: during during acclimatation period (study week 2), and near the end of the treatment period (study week 12)
- Dose groups that were examined: all the animals
All ocular examinations were conducted using an indirect ophthalmoscope and slit lamp biomicroscope preceded by pupillary dilation with an appropriate mydriatic agent.

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at the scheduled necropsy
- Anaesthetic used for blood collection: Yes, isoflurane
- Animals fasted: Yes
- How many animals: all
- Parameters listed in table 1 were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at the scheduled necropsy
- Animals fasted: Yes
- How many animals: all
- Parameters listed in table 2 were examined.

URINALYSIS: Yes
- Time schedule for collection of urine: at the scheduled necropsy
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes, overnight
- Parameters listed in table 3 were examined.

NEUROBEHAVIOURAL EXAMINATION: No

Sacrifice and pathology:
GROSS PATHOLOGY: Yes, on all animals

HISTOPATHOLOGY: Yes, on all animals
Statistics:
All statistical tests were performed using WTDMS™. Analyses were conducted using two-tailed tests for minimum significance levels of 1% and 5%, comparing each test substance-treated group to the control group by sex. Body weight, body weight change, food consumption, clinical pathology and organ weight (absolute and relative) data were subjected to a parametric one way ANOVA (Snedecor and Cochran, 1980) to determine intergroup differences. If the ANOVA revealed statistically significant (p<0.05) intergroup variance, Dunnett's test (Dunnett, 1964) was used to compare the test substance treated groups to the control group.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
1 control group male found dead (day 37), one 280 ppm group male (day 90), and one 1000 ppm group male (day 92, day of scheduled necropsy). 1 control group male (day 12) was replaced. One 280 ppm group male (day 11) was replaced
Mortality:
mortality observed, treatment-related
Description (incidence):
1 control group male found dead (day 37), one 280 ppm group male (day 90), and one 1000 ppm group male (day 92, day of scheduled necropsy). 1 control group male (day 12) was replaced. One 280 ppm group male (day 11) was replaced
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
small effect on mean body weight in the 1000 ppm group females. No effect in males.
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:
no effects observed
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
no effects observed
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
Higher mean kidney and liver weights (relative to final body weight) in the 1000 ppm females, but not correlated with microscopic findings and considered to be a result of a non-statistically significant, test substance-related effect on final body weight
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
effects observed, treatment-related
Description (incidence and severity):
malignant lymphoma in 1 mid-dose animal, considered unrelated to treatment
Details on results:
CLINICAL SIGNS AND MORTALITY
Mortality: 3 male animals found dead during the study. 1 control group male found dead on study day 37, one 280 ppm group male found dead on study day 90, and one 1000 ppm group male found dead on study day 92 (day of scheduled necropsy). The cause of death of these animals was undetermined. Deaths occurred at different days and in different groups, including one animal in the control group, and lack of histologic correlates led to conclude that the deaths were not test substance related. In addition, 2 males were replaced after being found dead on study days 12 and 11 (control and 280 ppm group, respectively); however, no clinical findings were noted for the 280 ppm group male that would be associated with test substance exposure.
Clinical signs: yellow material around the urogenital area in the 280 and 1000 ppm group females beginning (from day 14 throughout study period). Red material around the nose noted in the 1000 ppm group males and females at 1 hour post-exposure throughout the study period. However, this observation was considered transient and not toxicologically significant.

BODY WEIGHT:
In females, there was a small effect on mean body weight in the 1000 ppm group. Although not statistically significant, body weight was 8% lower than the control group mean following 12 and 13 weeks of exposure. Mean cumulative body weight gain for this group was 16.5% lower than control group (study week 13). In males, body weights were unaffected by test substance exposure.

HAEMATOLOGY:
There were no test substance-related alterations in hematology and coagulation parameters.
However, some statistically significant differences were observed when the control and test substance-treated groups were compared, however without an exposure-related response (increased WBC and lymphocyte counts in the 280 ppm group females and decreased platelet counts in the 280 ppm group males).
The decreased group mean value for the platelet count in this group was mainly due to 1 animal which presented with malignant lymphoma and was not test substance related.

CLINICAL CHEMISTRY:
There were no test substance-related alterations in serum chemistry parameters.
However, some statistically significant differences were observed when the control and test substance treated groups were compared. The mean cholesterol value was higher in the 1000 ppm group females (+27.8%) than in the control group. However, the value was within the range of values in the laboratory historical control database and lacked correlating histologic findings or changes in other related serum chemistry parameters, and therefore was considered the result of biologic variation

ORGAN WEIGHTS:
Higher kidney (+12.9%) and liver weights (+14.9%) (relative to final body weight) in the 1000 ppm group females were statistically significant when compared to the control group, but were not correlated with histologic findings and were considered to be a result of a non-statistically significant, test substance-related effect on final body weight.
There were no other test substance-related effects on organ weights. However, some statistically significant differences were observed when the control and test substance treated groups were compared. The adrenal gland weight relative to brain weight was lower for the 1000 ppm group females than the control group, but lacked correlating histologic changes and was within the range of values in the laboratory historical control database, and therefore was considered the result of biologic variability. Spleen weights (absolute and relative to final body weight) were higher in the 280 ppm group females than the control group females. In the absence of an exposure-relationship or correlating histologic findings and because the values were within the range of values in the laboratory historical control database, the higher spleen weights were not considered test substance-related.

GROSS PATHOLOGY:
no observations that were considered to be associated with administration of the test substance.

HISTOPATHOLOGY: NON-NEOPLASTIC
There were no test substance-related microscopic findings noted in this study.
All histologic changes were considered to be incidental findings or related to some aspect of experimental manipulation other than administration of the test substance. There was no test substance related alteration in the prevalence, severity, or histologic character of those incidental tissue alterations.

HISTOPATHOLOGY: NEOPLASTIC
One male in the 280 ppm group presented with malignant lymphoma at the scheduled necropsy in the mandibular, mediastinal, and mesenteric lymph nodes, spleen, thymus, lungs, and liver, but was not test-article related because the neoplasm occurred in only 1 animal in the mid-exposure group.
Dose descriptor:
NOAEC
Effect level:
ca. 1 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: overall effects
Critical effects observed:
not specified

 

Selected Clinical Observations

Sex

Males

Females

Exposure Level (ppm)

0

80

280

1000

0

80

280

1000

Observationa

 

Dried red material around nose

 

 

 

 

1 Hour Post-Exposure

2/2

3/3

4/3

12/5

0/0

3/3

7/4

24/8

Wet yellow material urogenital area

 

 

 

 

1 Hour Post-Exposure

0/0

0/0

0/0

0/0

0/0

0/0

20/3

109/7

Dried yellow material urogenital area

 

 

 

 

Detailed Physicals

0/0

0/0

0/0

0/0

0/0

0/0

0/0

15/4

Prior to Exposure

0/0

0/0

0/0

0/0

0/0

0/0

0/0

50/4

1 Hour Post-Exposure

0/0

0/0

0/0

0/0

0/0

0/0

3/2

21/6

Non-Exposure Days

0/0

0/0

0/0

0/0

0/0

0/0

1/1

12/3

 a= Observations reported as total occurrence/number of animals.

Conclusions:
Administration of PMVE via whole-body inhalation for 6 hours per day on a 5-day per week basis for 13 weeks to Sprague-Dawley rats at exposure levels up to 1000 ppm was well tolerated. Non-adverse test substance-related effects were limited to clinical observations of yellow material around the urogenital area in the 280 and 1000 ppm group females and slightly lower mean body weights in the 1000 ppm group females. There was no remarkable histopathological findings. Therefore, the no observed adverse effect concentration (NOAEC) was 1000 ppm.
Executive summary:

The potential toxic effects of trifluoro(trifluoromethoxy)ethylene (also referred to as PMVE) were assessed in Sprague Dawley rats administered the substance via whole-body inhalation for 6 hours per day on a 5-day per week basis for 13 weeks (minimum of 65 exposures for each animal).

Trifluoro(trifluoromethoxy)ethylene was administered to 3 groups (Groups 2‑4) of Crl:CD(SD) rats. Target exposure concentrations were 80, 280, and 1000 ppm for Groups 2, 3, and 4, respectively. Analyzed exposure concentrations were 81, 279, and 1004 ppm for Groups 2, 3, and 4, respectively. A concurrent control group (Group 1) was exposed to humidified, filtered air on a comparable regimen. Each group (Groups 1-4) consisted of 10 animals/sex. Following 13 weeks of exposure, all rats/sex/group were euthanized. All standard examinations were performed. Intergroup differences were assessed using parametric one-way ANOVA test, and, when appropriate, Dunnett's test was used to compare treated groups to the control group.

There were no test substance-related effects on survival, food consumption, clinical pathology parameters, organ weights, macroscopic observations, or microscopic findings. In addition, there were no test substance-related ophthalmic lesions indicative of toxicity (examinations performed during study weeks -2 and 12).

There were 3 male animals found dead during the study: 1 control group male found dead on day 37, one 280 ppm group male found dead on day 90, and one 1000 ppm group male found dead on day 92 (day of scheduled necropsy). The cause of death of these animals was undetermined; however, the occurrence at different days and in different groups, including an animal in the control group, and lack of histologic correlates led to the determination that the deaths were not test substance‑related. In addition, 2 males were replaced after being found dead on study days 12 and 11 (control and 280 ppm group, respectively); however, no clinical findings were noted for the 280 ppm group male that would be associated with test substance exposure.

Test substance-related clinical observations of yellow material around the urogenital area were noted in the 280 and 1000 ppm group females beginning as early as study day 14 and continuing throughout study period. The test substance-related observation of red material around the nose was noted in the 1000 ppm group males and females at 1 hour post-exposure throughout the study period. However, this observation was considered transient and not toxicologically significant. In females, test substance exposure resulted in a small effect on mean body weight in the 1000 ppm group. Body weight was 8% lower than the control group mean following 12 and 13 weeks of exposure. Compared to control group mean values, higher mean kidney and liver weights (relative to final body weight) were noted in the 1000 ppm group females, but were not correlated with microscopic findings and were considered to be a result of a non-statistically significant, test substance-related effect on final body weight.

Administration of PMVE via whole-body inhalation for 6 hours per day on a 5-day per week basis for 13 weeks to Crl:CD(SD) rats at exposure levels up to 1000 ppm was well tolerated. Non-adverse test substance-related effects were limited to clinical observations of yellow material around the urogenital area in the 280 and 1000 ppm group females and slightly lower mean body weights in the 1000 ppm group females. Therefore, the no‑observed‑adverse‑effect concentration (NOAEC) was 1000 ppm.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEC
6 790.26 mg/m³
Study duration:
subchronic
Species:
rat
Quality of whole database:
Consistent results were obtained in two good quality repeated dose studies.

Repeated dose toxicity: inhalation - local effects

Link to relevant study records
Reference
Endpoint:
sub-chronic toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2015-04-22 to 2016-05-06
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study
Qualifier:
according to
Guideline:
OECD Guideline 413 (Subchronic Inhalation Toxicity: 90-Day Study)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.29 (Sub-Chronic Inhalation Toxicity:90-Day Study)
Deviations:
no
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Crj: CD(SD)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Inc., Raleigh, NC
- Age at study initiation: 38 days old at receipt, 7 weeks old at the initiation of test substance exposures
- Weight at study initiation: Individual body weights ranged from 236 g to 283 g for males and from 172 g to 214 g for females at the initiation of test substance exposures
- Housing: 2 or 3 per cage
- Diet : PMI Nutrition International, LLC, Certified Rodent LabDiet® 5002 (meal), ad libitum, except during the exposure periods and except during the period of fasting prior to necropsy
- Water : Reverse osmosis-treated (on-site) drinking water, delivered by an automatic watering system. Ad libitum except during the exposure periods.
- Acclimation period: 14 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Actual mean daily temperature ranged from 19.0°C to 22.7°C during the study
- Humidity (%): actual mean daily relative humidity ranged from 43.0% to 55.8% during the study
- Air changes (per hr): minimum of 10 fresh air changes per hour
- Photoperiod : 12 hour light (0600 hours to 1800 hours)/12 hour dark

IN-LIFE DATES: From: 07May2015 To: 06Aug2015
Route of administration:
inhalation: gas
Type of inhalation exposure:
whole body
Vehicle:
air
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: 4 approximately 1000-L stainless steel and glass whole body inhalation exposure chambers
- Method of holding animals in test chamber: whole body inhalation exposure chambers
- source of air: filtered air
- Method of conditioning air: A HEPA-filter and an activated-charcoal bed were used to pre-treat room air prior to delivery to chambers.
- Temperature, humidity, pressure in air chamber: mean temperature 19 to 25°C, mean humidity 30 to 70%, under slight negative pressure.
- Air flow rate: 200 to 250 Standard liters per minute
- Air change rate: 12 to 15 ACH
- Treatment of exhaust air: Chamber exhaust was passed through the facility exhaust system consisting of redundant exhaust blowers preceded by activated-charcoal and HEPA-filter units.

TEST ATMOSPHERE
- Brief description of analytical method used: Analyzed exposure concentrations were determined at approximately 45-minute intervals using an appropriate gas chromatography (GC) method
- Samples taken from breathing zone: yes

CONTROL GROUP
Concurrent exposure to humidified, filtered air on a comparable regimen
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The test substance usage was documented by weighing the gas cylinder prior and following exposure.
Analyzed exposure concentrations were determined at approximately 45-minute intervals using an appropriate gas chromatography (GC) method after calibration with standards prepared at 5 gas concentrations. The first sampling round initiated approximately 18 min after the start of exposure atmosphere generation.
- Homogeneity was assessed during method development: 4 remote locations.

Duration of treatment / exposure:
6 hours per day on a 5-day per week basis for 13 weeks (65 total exposures)
Frequency of treatment:
5-day per week
Remarks:
Doses / Concentrations:
0, 80, 280, and 1000 ppm
Basis:
nominal conc.
Remarks:
Doses / Concentrations:
0, 81, 279, and 1004 ppm
Basis:
analytical conc.
No. of animals per sex per dose:
10 animals
Control animals:
other: humidified, filtered air on a comparable regimen
Details on study design:
- Dose selection rationale: based on results of an OECD 422 study
Positive control:
No
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily for mortality and moribundity

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: detailed physical examinations were performed weekly

BODY WEIGHT: Yes
- Time schedule for examinations: Individual body weights were recorded twice weekly (± 1 day) during the first 4 weeks of the exposure period, then weekly for the remainder of the study.

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
Food consumption was recorded once weekly (± 2 days) beginning after randomization

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: during during acclimatation period (study week 2), and near the end of the treatment period (study week 12)
- Dose groups that were examined: all the animals
All ocular examinations were conducted using an indirect ophthalmoscope and slit lamp biomicroscope preceded by pupillary dilation with an appropriate mydriatic agent.

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at the scheduled necropsy
- Anaesthetic used for blood collection: Yes, isoflurane
- Animals fasted: Yes
- How many animals: all
- Parameters listed in table 1 were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at the scheduled necropsy
- Animals fasted: Yes
- How many animals: all
- Parameters listed in table 2 were examined.

URINALYSIS: Yes
- Time schedule for collection of urine: at the scheduled necropsy
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes, overnight
- Parameters listed in table 3 were examined.

NEUROBEHAVIOURAL EXAMINATION: No

Sacrifice and pathology:
GROSS PATHOLOGY: Yes, on all animals

HISTOPATHOLOGY: Yes, on all animals
Statistics:
All statistical tests were performed using WTDMS™. Analyses were conducted using two-tailed tests for minimum significance levels of 1% and 5%, comparing each test substance-treated group to the control group by sex. Body weight, body weight change, food consumption, clinical pathology and organ weight (absolute and relative) data were subjected to a parametric one way ANOVA (Snedecor and Cochran, 1980) to determine intergroup differences. If the ANOVA revealed statistically significant (p<0.05) intergroup variance, Dunnett's test (Dunnett, 1964) was used to compare the test substance treated groups to the control group.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
1 control group male found dead (day 37), one 280 ppm group male (day 90), and one 1000 ppm group male (day 92, day of scheduled necropsy). 1 control group male (day 12) was replaced. One 280 ppm group male (day 11) was replaced
Mortality:
mortality observed, treatment-related
Description (incidence):
1 control group male found dead (day 37), one 280 ppm group male (day 90), and one 1000 ppm group male (day 92, day of scheduled necropsy). 1 control group male (day 12) was replaced. One 280 ppm group male (day 11) was replaced
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
small effect on mean body weight in the 1000 ppm group females. No effect in males.
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:
no effects observed
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
no effects observed
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
Higher mean kidney and liver weights (relative to final body weight) in the 1000 ppm females, but not correlated with microscopic findings and considered to be a result of a non-statistically significant, test substance-related effect on final body weight
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
effects observed, treatment-related
Description (incidence and severity):
malignant lymphoma in 1 mid-dose animal, considered unrelated to treatment
Details on results:
CLINICAL SIGNS AND MORTALITY
Mortality: 3 male animals found dead during the study. 1 control group male found dead on study day 37, one 280 ppm group male found dead on study day 90, and one 1000 ppm group male found dead on study day 92 (day of scheduled necropsy). The cause of death of these animals was undetermined. Deaths occurred at different days and in different groups, including one animal in the control group, and lack of histologic correlates led to conclude that the deaths were not test substance related. In addition, 2 males were replaced after being found dead on study days 12 and 11 (control and 280 ppm group, respectively); however, no clinical findings were noted for the 280 ppm group male that would be associated with test substance exposure.
Clinical signs: yellow material around the urogenital area in the 280 and 1000 ppm group females beginning (from day 14 throughout study period). Red material around the nose noted in the 1000 ppm group males and females at 1 hour post-exposure throughout the study period. However, this observation was considered transient and not toxicologically significant.

BODY WEIGHT:
In females, there was a small effect on mean body weight in the 1000 ppm group. Although not statistically significant, body weight was 8% lower than the control group mean following 12 and 13 weeks of exposure. Mean cumulative body weight gain for this group was 16.5% lower than control group (study week 13). In males, body weights were unaffected by test substance exposure.

HAEMATOLOGY:
There were no test substance-related alterations in hematology and coagulation parameters.
However, some statistically significant differences were observed when the control and test substance-treated groups were compared, however without an exposure-related response (increased WBC and lymphocyte counts in the 280 ppm group females and decreased platelet counts in the 280 ppm group males).
The decreased group mean value for the platelet count in this group was mainly due to 1 animal which presented with malignant lymphoma and was not test substance related.

CLINICAL CHEMISTRY:
There were no test substance-related alterations in serum chemistry parameters.
However, some statistically significant differences were observed when the control and test substance treated groups were compared. The mean cholesterol value was higher in the 1000 ppm group females (+27.8%) than in the control group. However, the value was within the range of values in the laboratory historical control database and lacked correlating histologic findings or changes in other related serum chemistry parameters, and therefore was considered the result of biologic variation

ORGAN WEIGHTS:
Higher kidney (+12.9%) and liver weights (+14.9%) (relative to final body weight) in the 1000 ppm group females were statistically significant when compared to the control group, but were not correlated with histologic findings and were considered to be a result of a non-statistically significant, test substance-related effect on final body weight.
There were no other test substance-related effects on organ weights. However, some statistically significant differences were observed when the control and test substance treated groups were compared. The adrenal gland weight relative to brain weight was lower for the 1000 ppm group females than the control group, but lacked correlating histologic changes and was within the range of values in the laboratory historical control database, and therefore was considered the result of biologic variability. Spleen weights (absolute and relative to final body weight) were higher in the 280 ppm group females than the control group females. In the absence of an exposure-relationship or correlating histologic findings and because the values were within the range of values in the laboratory historical control database, the higher spleen weights were not considered test substance-related.

GROSS PATHOLOGY:
no observations that were considered to be associated with administration of the test substance.

HISTOPATHOLOGY: NON-NEOPLASTIC
There were no test substance-related microscopic findings noted in this study.
All histologic changes were considered to be incidental findings or related to some aspect of experimental manipulation other than administration of the test substance. There was no test substance related alteration in the prevalence, severity, or histologic character of those incidental tissue alterations.

HISTOPATHOLOGY: NEOPLASTIC
One male in the 280 ppm group presented with malignant lymphoma at the scheduled necropsy in the mandibular, mediastinal, and mesenteric lymph nodes, spleen, thymus, lungs, and liver, but was not test-article related because the neoplasm occurred in only 1 animal in the mid-exposure group.
Dose descriptor:
NOAEC
Effect level:
ca. 1 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: overall effects
Critical effects observed:
not specified

 

Selected Clinical Observations

Sex

Males

Females

Exposure Level (ppm)

0

80

280

1000

0

80

280

1000

Observationa

 

Dried red material around nose

 

 

 

 

1 Hour Post-Exposure

2/2

3/3

4/3

12/5

0/0

3/3

7/4

24/8

Wet yellow material urogenital area

 

 

 

 

1 Hour Post-Exposure

0/0

0/0

0/0

0/0

0/0

0/0

20/3

109/7

Dried yellow material urogenital area

 

 

 

 

Detailed Physicals

0/0

0/0

0/0

0/0

0/0

0/0

0/0

15/4

Prior to Exposure

0/0

0/0

0/0

0/0

0/0

0/0

0/0

50/4

1 Hour Post-Exposure

0/0

0/0

0/0

0/0

0/0

0/0

3/2

21/6

Non-Exposure Days

0/0

0/0

0/0

0/0

0/0

0/0

1/1

12/3

 a= Observations reported as total occurrence/number of animals.

Conclusions:
Administration of PMVE via whole-body inhalation for 6 hours per day on a 5-day per week basis for 13 weeks to Sprague-Dawley rats at exposure levels up to 1000 ppm was well tolerated. Non-adverse test substance-related effects were limited to clinical observations of yellow material around the urogenital area in the 280 and 1000 ppm group females and slightly lower mean body weights in the 1000 ppm group females. There was no remarkable histopathological findings. Therefore, the no observed adverse effect concentration (NOAEC) was 1000 ppm.
Executive summary:

The potential toxic effects of trifluoro(trifluoromethoxy)ethylene (also referred to as PMVE) were assessed in Sprague Dawley rats administered the substance via whole-body inhalation for 6 hours per day on a 5-day per week basis for 13 weeks (minimum of 65 exposures for each animal).

Trifluoro(trifluoromethoxy)ethylene was administered to 3 groups (Groups 2‑4) of Crl:CD(SD) rats. Target exposure concentrations were 80, 280, and 1000 ppm for Groups 2, 3, and 4, respectively. Analyzed exposure concentrations were 81, 279, and 1004 ppm for Groups 2, 3, and 4, respectively. A concurrent control group (Group 1) was exposed to humidified, filtered air on a comparable regimen. Each group (Groups 1-4) consisted of 10 animals/sex. Following 13 weeks of exposure, all rats/sex/group were euthanized. All standard examinations were performed. Intergroup differences were assessed using parametric one-way ANOVA test, and, when appropriate, Dunnett's test was used to compare treated groups to the control group.

There were no test substance-related effects on survival, food consumption, clinical pathology parameters, organ weights, macroscopic observations, or microscopic findings. In addition, there were no test substance-related ophthalmic lesions indicative of toxicity (examinations performed during study weeks -2 and 12).

There were 3 male animals found dead during the study: 1 control group male found dead on day 37, one 280 ppm group male found dead on day 90, and one 1000 ppm group male found dead on day 92 (day of scheduled necropsy). The cause of death of these animals was undetermined; however, the occurrence at different days and in different groups, including an animal in the control group, and lack of histologic correlates led to the determination that the deaths were not test substance‑related. In addition, 2 males were replaced after being found dead on study days 12 and 11 (control and 280 ppm group, respectively); however, no clinical findings were noted for the 280 ppm group male that would be associated with test substance exposure.

Test substance-related clinical observations of yellow material around the urogenital area were noted in the 280 and 1000 ppm group females beginning as early as study day 14 and continuing throughout study period. The test substance-related observation of red material around the nose was noted in the 1000 ppm group males and females at 1 hour post-exposure throughout the study period. However, this observation was considered transient and not toxicologically significant. In females, test substance exposure resulted in a small effect on mean body weight in the 1000 ppm group. Body weight was 8% lower than the control group mean following 12 and 13 weeks of exposure. Compared to control group mean values, higher mean kidney and liver weights (relative to final body weight) were noted in the 1000 ppm group females, but were not correlated with microscopic findings and were considered to be a result of a non-statistically significant, test substance-related effect on final body weight.

Administration of PMVE via whole-body inhalation for 6 hours per day on a 5-day per week basis for 13 weeks to Crl:CD(SD) rats at exposure levels up to 1000 ppm was well tolerated. Non-adverse test substance-related effects were limited to clinical observations of yellow material around the urogenital area in the 280 and 1000 ppm group females and slightly lower mean body weights in the 1000 ppm group females. Therefore, the no‑observed‑adverse‑effect concentration (NOAEC) was 1000 ppm.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed

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

Additional information

As the test substance is a gas at room temperature, the test is not technically feasible through oral and dermal routes; in addition, inhalation exposure is the most relevant route of exposure . In fact, detailed data via the inhalation route is available. A GLP test (Report No. DUPONT-20813, 2007) following OECD guideline 422 showed that exposure to the test substance did not result in adverse clinical signs or mortality. Test substance-related reductions in weight gain, food consumption, and/or food efficiency occurred in 1500ppm males and females; however, they were transient and did not adversely affect the health of the animals. There were no adverse or test substance-related effects on neurobehavioral parameters, clinical pathology parameters, and no effects on clinical observations, or survival. Test substance-related, minimal regeneration of renal tubular epithelium was observed in 1500ppm males and females, and was accompanied by increased absolute and relative kidney weights in 1500ppm females. Based on the results above, the NOEC for systemic toxicity was 300ppm ( 2037,08 mg/m3).

In the follow-up study, a GLP 90-day inhalation study in rats conducted at the doses 0, 80, 280 and 1000 ppm, only few clinical signs were observed, and were limited to non-adverse test substance-related effects consisting of yellow material around the urogenital area in the 280 and 1000 ppm group females and slightly lower mean body weights in the 1000 ppm group females. There were no remarkable histopathological findings in males and females at the highest dose. Therefore, the no observed adverse effect concentration (NOAEC) was 1000 ppm (6790.26 mg/m3).

 


Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
In accordance with section 1 of REACH (Regulation (EC) No 1907/2006) Annex XI the oral repeated toxicity study (required in section 8.6.1 Annex VIII) does not need to be conducted as inhalation exposure is the most relevant route of exposure. In addition PMVE is a gas at standard temperature and pressure consequently the oral toxicity test is technically not feasible.

Justification for selection of repeated dose toxicity inhalation - systemic effects endpoint:
Adverse effects were initially observed in the 28-day (OECD422) at 1500 ppm, while in the 90-day study despite a longer exposure period the highest dose 1000 ppm could be considered a NOAEC. The sub-chronic study performed according to the OECD413 Guideline and GLP was therefore selected as key study and starting point for the DNEL derivation for long-term systemic effects by inhalation.

Justification for selection of repeated dose toxicity inhalation - local effects endpoint:
guideline study following GLP. There were no specific observations indicating local effects on the respiratory tract in the repeated dose-toxicity study in rats.

Justification for selection of repeated dose toxicity dermal - systemic effects endpoint:
In accordance with section 1 of REACH (Regulation (EC) No 1907/2006) Annex XI the dermal repeated toxicity study (required in section 8.6.1 Annex VIII) does not need to be conducted as inhalation exposure is the most relevant route of exposure. In addition PMVE is a gas at standard temperature and pressure, consequently the dermal toxicity test is technically not feasible.

Justification for selection of repeated dose toxicity dermal - local effects endpoint:
In accordance with section 1 of REACH (Regulation (EC) No 1907/2006) Annex XI the dermal repeated toxicity study (required in section 8.6.1 Annex VIII) does not need to be conducted as inhalation exposure is the most relevant route of exposure. In addition PMVE is a gas at standard temperature and pressure, consequently the dermal toxicity test is technically not feasible..

Repeated dose toxicity: inhalation - systemic effects (target organ) urogenital: kidneys

Justification for classification or non-classification

Based on the results of the study, the No-Observed-Adverse-Effect Concentration (NOAEC) for systemic toxicity was 1000 ppm (6790.26 mg/m3). PMVE does not meet the classification criteria for hazard classes related to repeated dose exposure according to Regulation (EC) 1272/2008.