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EC number: - | CAS number: -
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Acute Toxicity: inhalation
Administrative data
- Endpoint:
- acute toxicity: inhalation
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- The study was conducted between 29 June 2017 and 05 October 2017.
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 018
- Report date:
- 2018
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 403 (Acute Inhalation Toxicity)
- Deviations:
- yes
- Remarks:
- The deviation was considered to have not affected the integrity or validity of the study.
- GLP compliance:
- yes (incl. QA statement)
- Test type:
- acute toxic class method
- Limit test:
- no
Test material
- Reference substance name:
- bis(cyclohexylmethyl) ether
- Cas Number:
- 14315-63-0
- Molecular formula:
- C14H26O
- IUPAC Name:
- bis(cyclohexylmethyl) ether
- Test material form:
- liquid
- Remarks:
- Clear, colorless
Constituent 1
- Specific details on test material used for the study:
- Identification: FRET 13-0460
Physical state/appearance: clear colorless liquid
Storage Conditions: approximately 4 °C in the dark
Test animals
- Species:
- rat
- Strain:
- Wistar
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- Animal Information
Male and female RccHan™ : WIST strain rats were supplied by Envigo RMS (UK) Limited, Oxon, UK. On receipt the animals were randomly allocated to cages. After an acclimatization period of at least 5 days the animals were given a number unique within the study by ear punching and a number written on a color coded cage card. At the start of the study the animals were approximately 8 to 12 weeks old and within the weight range of 200 g to 350 g. The females were nulliparous and non pregnant.
Animal Care and Husbandry
The animals were housed in groups of up to five by sex in solid floor polypropylene cages with stainless steel lids, furnished with softwood flakes. With the exception of the exposure period, free access to mains drinking water and food (2014C Teklad Global Rodent diet supplied by Envigo RMS (UK) Limited, Oxon, UK) was allowed throughout the study. The diet, drinking water and bedding were routinely analyzed and were considered not to contain any contaminants that would reasonably be expected to affect the purpose or integrity of the study.
The temperature and relative humidity were set to achieve limits of 19 to 25 °C and 30 to 70% respectively. The rate of air exchange was at least fifteen changes per hour and the lighting was controlled by a time switch to give 12 hours continuous light and 12 hours darkness. The animals were retained in this accommodation at all times except during the exposure period.
The animals were provided with environmental enrichment items which were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.
Administration / exposure
- Route of administration:
- inhalation: aerosol
- Type of inhalation exposure:
- nose only
- Vehicle:
- air
- Mass median aerodynamic diameter (MMAD):
- 2.65 µm
- Geometric standard deviation (GSD):
- 2.28
- Details on inhalation exposure:
- Atmosphere Generation
The test item was aerosolized using a metal concentric jet nebulizer (Envigo CRS Limited, UK) located at the top of the exposure chamber. The nebulizer was connected to a glass syringe attached to an infusion pump, which provided a continuous supply of test item under pressure, and to a metered compressed air supply.
Compressed air was supplied by means of an oil free compressor and passed through a water trap and respiratory quality filters before it was introduced to the nebulizer.
The cylindrical exposure chamber had a volume of approximately 30 liters (dimensions: 28 cm diameter x 50 cm high). The concentration within the exposure chamber was controlled by adjusting the rate of the infusion pump. The extract from the exposure chamber passed through a ‘scrubber’ trap and was connected with a high efficiency filter to a metered exhaust system. The chamber was maintained under negative pressure.
Homogeneity of the test atmosphere within the chamber was not specifically determined during this study. Chambers of the same design (ADG Developments Ltd, Hitchin, Herts, UK) have been fully validated and shown to produce evenly distributed atmospheres in the animals’ breathing zone with a wide variety of test items (Green J D et al, 1984).
Prior to the start of the study, test item atmospheres were generated within the exposure chamber. During this characterization period test item input rates were varied to achieve the required atmospheric conditions.
Exposure Procedure
During the exposure period, each rat was individually held in a tapered, polycarbonate restraining tube fitted onto a single tier of the exposure chamber and sealed by means of a rubber ‘O’ ring. Only the nose of each animal was exposed to the test atmosphere.
Following an appropriate equilibration period a single group of ten rats (five males and five females), was subjected to a single exposure to the test item for a period of four hours. A target concentration of 5.0 mg/L was used for the exposure. As the mean achieved concentration was 102% of target and no deaths occurred, no further levels were required.
Exposure Chamber Temperature and Relative Humidity
The temperature and relative humidity inside the exposure chamber were measured by an electronic thermometer/humidity meter (Hanna Instruments Ltd, Beds., UK) located in a vacant port in the animals’ breathing zone of the chamber and recorded every 30 minutes throughout the 4 Hour exposure period.
Exposure Chamber Oxygen Concentration
Oxygen levels within the exposure chamber were measured by an electronic oxygen analyzer (Servomex (UK) Ltd, Crowborough, East Sussex) located in a port in the animals breathing zone during the 4 Hour exposure period. The test atmospheres were generated to contain at least 19% oxygen.
Exposure Chamber Atmosphere Concentration
Prior to the inhalation phase of the study, the non-volatile component of the test item was determined by adding a small, known amount of test item to glass fiber filters and recording their weights. The filters were then dried in a desiccator at room temperature for approximately 24 hours and then weighed again. The difference in the two weights was taken as the volatile content of the test item and the non-volatile component was calculated as a percentage. The mean non-volatile component of the batch used during the formal exposure was found to be 69.78 % (n=8). A significant difference between the expected and achieved concentrations was noted during generation trials therefore it was considered that gravimetric analysis would not be accurate and that chemical analysis would be required.
The test atmosphere was sampled nine times during the exposure period. A weighed glass fiber filter was placed in a filter holder and temporarily sealed in a vacant port of the exposure chamber in the animals’ breathing zone. A known quantity of the exposure chamber atmosphere was drawn through the filter using a vacuum pump. The samples were then submitted for chemical analysis.
The nominal chamber concentration was calculated by dividing the mass of test item disseminated into the chamber by the total volume of air that flowed through the chamber during the exposure.
The nominal concentration was 373% of the actual mean achieved atmosphere concentration and shows that keeping the aerosol airborne was relatively straightforward.
Particle Size Distribution
The particle size of the generated atmosphere inside the exposure chamber was determined three times during the exposure period using a Marple Personal Cascade Impactor (Westech IS Ltd, Beds., UK). This device consisted of six impactor stages (10.4, 7.7, 4.1, 1.3, 0.90 and 0.56 µm cut points) with stainless steel collection substrates and a backup glass fiber filter, housed in an aluminum sampler. The sampler was temporarily sealed in a sampling port in the animals’ breathing zone and a suitable, known volume of exposure chamber air was drawn through it using a vacuum pump.
The collection substrates and backup filter were weighed before and after sampling and the weight of test item, collected at each stage, calculated by difference.
The mean amount for each stage was used to determine the cumulative amount below each cut-off point size. In this way, the proportion (percentage) of aerosol less than 10.4, 7.7, 4.1, 1.3, 0.90 and 0.56 µm was calculated.
The resulting values were converted to probits and plotted against Log10 cut point size. From this plot, the Mass Median Aerodynamic Diameter (MMAD) was determined (as the 50% point) and the geometric standard deviation was calculated. In addition the proportion (percentage) of aerosol less than 4 µm (considered to be the inhalable fraction) was determined. - Analytical verification of test atmosphere concentrations:
- yes
- Duration of exposure:
- 4 h
- Concentrations:
- A target concentration of 5.0 mg/L was used for the exposure.
- No. of animals per sex per dose:
- 5 males and 5 females
- Control animals:
- no
- Details on study design:
- Serial Observations
Clinical Signs
All animals were observed for clinical signs at hourly intervals during exposure, immediately on removal from the restraining tubes at the end of exposure, 1 hour after termination of exposure and subsequently once daily for 14 days. Any evidence of overt toxicity was recorded at each observation.
Body Weight
Individual body weights were recorded on arrival, prior to treatment on the day of exposure (Day 0) and on Days 1, 3, 7 and 14.
Terminal Investigations
Necropsy
All animals were subjected to a full external and internal examination and any macroscopic abnormalities were recorded. The respiratory tract was subjected to a detailed macroscopic examination for signs of irritancy or local toxicity.
Data Evaluation
Data evaluations included the relationship, if any, between the animals’ exposure to the test item and the incidence and severity of all abnormalities including behavioral and clinical observations, necropsy findings, body weight changes, mortality and any other toxicological effects.
Using the mortality data obtained, an estimate of the acute inhalation median lethal concentration (LC50) of the test item was made.
Results and discussion
Effect levels
- Sex:
- male/female
- Dose descriptor:
- LC50
- Effect level:
- > 5.11 mg/L air
- Based on:
- test mat.
- Exp. duration:
- 4 h
- Mortality:
- No mortality was observed.
- Clinical signs:
- other: Signs of hunched posture and pilo-erection are commonly seen in animals for short periods on removal from the chamber following 4-hour inhalation studies. Wet fur is commonly recorded both during and for a short period after exposure. These observations
- Body weight:
- All animals exhibited body weight loss or no body weight gain on the first day post-exposure. With the exception of one female from Days 1 to 3 post-exposure and three females from Days 3 to 7 post-exposure which exhibited body weight loss, body weight gains were noted for all animals during the remainder of the recovery period.
- Gross pathology:
- No macroscopic abnormalities were detected amongst animals at necropsy.
Any other information on results incl. tables
Exposure Chamber Concentration
The actual concentration of the test item was determined by gas chromatography (GC) using an external standard technique. The test atmosphere was sampled nine times and the actual concentration of the test item calculated. The mean values obtained were as follows:
Group Number |
Atmosphere Concentration |
||
Mean Achieved (mg/L) |
Standard Deviation |
Nominal (mg/L) |
|
1 |
5.11 |
0.29 |
19.04 |
The chamber flow rate was maintained at 40 L/min providing 80 air changes per hour.
The theoretical chamber equilibration time (T99) was 4 minutes[1](Silver, 1946).
Particle Size Distribution
The particle size analysis of the atmosphere drawn from the animals’ breathing zone was as follows:
Group Number |
Mean Achieved Atmosphere Concentration (mg/L) |
Mean Mass Median Aerodynamic Diameter (µm) |
Inhalable Fraction (% <4 µm) |
Geometric Standard Deviation |
1 |
5.11 |
2.65 |
69.2 |
2.28 |
[1]= The test atmosphere was generated for a total of 11 minutes prior to animal insertion to ensure the target test item concentration was being achieved.
Temperature and Relative Humidity in Exposure Chamber
Time |
Chamber Temperature (ºC) |
Chamber Relative Humidity (%) During Exposure |
0 |
21 |
41 |
30 |
21 |
40 |
60 |
21 |
43 |
90 |
21 |
41 |
120 |
21 |
41 |
150 |
21 |
44 |
180 |
21 |
44 |
210 |
21 |
42 |
240 |
21 |
42 |
Air Flow and Oxygen Concentration in Exposure Chamber
Time |
Air Flow (L/minute) |
Oxygen Concentration |
-11* |
40 |
- |
0 |
40 |
20.9 |
30 |
40 |
- |
60 |
40 |
- |
90 |
40 |
- |
120 |
40 |
20.9 |
150 |
40 |
- |
180 |
40 |
- |
210 |
40 |
- |
240 |
40 |
20.9 |
*= Test atmospheres were generated for a total of 11 minutes prior to animal insertion to ensure the target test item concentration was being achieved.
-= Not determined
Exposure Chamber Atmosphere Concentrations
Duration of Exposure (minutes) |
Volume of Air Sampled (L) |
Chamber Flow Rate (L/min) |
Atmosphere Concentration (mg/L) |
10 |
2 |
40 |
4.62 |
30 |
2 |
40 |
4.92 |
60 |
2 |
40 |
4.99 |
90 |
2 |
40 |
4.99 |
120 |
2 |
40 |
5.14 |
150 |
2 |
40 |
5.17 |
180 |
2 |
40 |
5.35 |
210 |
2 |
40 |
5.20 |
230 |
2 |
40 |
5.65 |
Mean achieved atmosphere concentration (mg/L) |
=5.11 |
|
Standard deviation |
=0.29 |
|
Nominal concentration: |
|
|
Test item used (g) |
191.2 |
|
Air flow (L/minute) |
40 |
|
Total generation time (minutes) |
251[1] |
|
Nominal concentration (mg/L) |
19.04 |
|
[1]= Test atmospheres were generated for a total of 11 minutes prior to animal insertion to ensure test item concentration was being achieved.
Particle Size Distribution
Cascade Impactor Data
Impactor Stage Number |
Cut Point |
Amount Collected (mg) per Sample Number |
Mean Amount Collected (mg) |
||
1 |
2 |
3 |
|||
3 |
10.4 |
0.05 |
0.12 |
0.09 |
0.09 |
4 |
7.7 |
0.13 |
0.18 |
0.15 |
0.15 |
5 |
4.1 |
0.35 |
0.43 |
0.35 |
0.38 |
6 |
1.3 |
0.49 |
0.60 |
0.55 |
0.55 |
7 |
0.90 |
0.49 |
0.54 |
0.41 |
0.48 |
8 |
0.56 |
0.18 |
0.16 |
0.10 |
0.15 |
Back-up Filter |
<0.56 |
0.00 |
0.08 |
0.00 |
0.03 |
Total Mean Amount of Test Item Collected |
1.83 |
Calculation
Cut Point (µm) |
Log10 |
Mean Cumulative Amount Less Than Cut Point |
||
(mg) |
(%) |
Probit |
||
10.4 |
1.017 |
1.74 |
95.1 |
6.65 |
7.7 |
0.887 |
1.59 |
86.9 |
6.12 |
4.1 |
0.613 |
1.21 |
66.1 |
5.42 |
1.3 |
0.114 |
0.66 |
36.1 |
4.64 |
0.90 |
-0.046 |
0.18 |
9.84 |
3.71 |
0.56 |
-0.252 |
0.03 |
1.64 |
2.87 |
Results |
|
|
|
Mean Mass Median Aerodynamic Diameter |
=2.65µm |
Geometric Standard Deviation |
=2.28 |
Predicted amount less than 4 µm |
=69.2% |
Mortality Data
Group Number |
Mean Achieved Atmosphere Concentration (mg/L) |
Sex |
Deaths During Exposure |
Deaths Post Exposure |
Deaths During Day of Observation |
Total Deaths |
|||||||
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8-14 |
||||||
1 |
5.11 |
Male |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0/10 |
Female |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
Individual Clinical Observations – (Day of Exposure)
Mean Achieved Atmosphere Concentration (mg/L) |
Animal Number and Sex |
Effects Noted During Exposure |
Effects Noted on Removal from Chamber |
Effects Noted 1 Hour Post Exposure |
||
1 |
2 |
3 |
||||
5.11 |
1 Male |
WfRd |
WfRd |
WfRd |
WfHPRd |
WfHPRd |
2 Male |
WfRd |
WfRd |
WfRd |
WfHPRd |
WfHPRd |
|
3 Male |
WfRd |
WfRd |
WfRd |
WfHPRd |
WfHPRd |
|
4 Male |
WfRd |
WfRd |
WfRd |
WfHPRd |
WfHPRd |
|
5 Male |
WfRd |
WfRd |
WfRd |
WfHPRd |
WfHPRd |
|
6 Female |
WfRd |
WfRd |
WfRd |
WfHPRd |
WfHPRd |
|
7 Female |
WfRd |
WfRd |
WfRd |
WfHPRd |
WfHPRd |
|
8 Female |
WfRd |
WfRd |
WfRd |
WfHPRdRs |
WfHPRd |
|
9 Female |
WfRd |
WfRd |
WfRd |
WfHPRdRs |
WfHPRd |
|
10 Female |
WfRd |
WfRd |
WfRd |
WfHPRd |
WfHPRd |
Wf = Wet fur
Rd = Decreased respiratory rate
H=Hunched posture
P = Pilo-erection
Rs = Sneezing
Individual Clinical Observations – Recovery Period
Mean Achieved Atmosphere Concentration (mg/L) |
Animal Number and Sex |
Effects Noted Post Exposure |
|||||||
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8-14 |
||
5.11 |
1 Male |
HP |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
2 Male |
HP |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
|
3 Male |
HP |
H |
0 |
0 |
0 |
0 |
0 |
0 |
|
4 Male |
HP |
0 |
Rn |
Rn |
Rn |
0 |
0 |
0 |
|
5 Male |
HPRnRs |
HRnRs |
0 |
0 |
0 |
0 |
0 |
0 |
|
6 Female |
HP |
H |
0 |
0 |
0 |
0 |
0 |
0 |
|
7 Female |
HP |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
|
8 Female |
HPRs |
HRs |
0 |
0 |
0 |
0 |
0 |
0 |
|
9 Female |
HP |
H |
0 |
0 |
0 |
0 |
0 |
0 |
|
10 Female |
HP |
H |
0 |
0 |
0 |
0 |
0 |
0 |
H=Hunched posture
P = Pilo-erection
Rn = Noisy respiration
Rs = Sneezing
0 = No abnormalities detected
Individual Body Weights and Body Weight Changes
Mean Achieved Atmosphere Concentration (mg/L) |
Animal Number and Sex |
Body Weight (g) at Day |
Body Weight Change (g) During Days |
|||||||||
-8 |
0 |
1 |
3 |
7 |
14 |
-8 to 0 |
0 to 1 |
1 to 3 |
3 to 7 |
7 to 14 |
||
5.11 |
1 Male |
202 |
248 |
234 |
245 |
259 |
291 |
46 |
-14 |
11 |
14 |
32 |
2 Male |
204 |
247 |
239 |
253 |
271 |
304 |
43 |
-8 |
14 |
18 |
33 |
|
3 Male |
206 |
250 |
237 |
245 |
263 |
303 |
44 |
-13 |
8 |
18 |
40 |
|
4 Male |
198 |
231 |
220 |
225 |
235 |
260 |
33 |
-11 |
5 |
10 |
25 |
|
5 Male |
202 |
245 |
237 |
241 |
264 |
295 |
43 |
-8 |
4 |
23 |
31 |
|
6 Female |
196 |
215 |
208 |
215 |
220 |
224 |
19 |
-7 |
7 |
5 |
4 |
|
7 Female |
204 |
210 |
207 |
213 |
211 |
219 |
6 |
-3 |
6 |
-2 |
8 |
|
8 Female |
207 |
222 |
222 |
221 |
226 |
243 |
15 |
0 |
-1 |
5 |
17 |
|
9 Female |
201 |
218 |
212 |
229 |
224 |
235 |
17 |
-6 |
17 |
-5 |
11 |
|
10 Female |
192 |
200 |
200 |
204 |
203 |
209 |
8 |
0 |
4 |
-1 |
6 |
Individual Necropsy Findings
Mean Achieved Atmosphere Concentration (mg/L) |
Animal Number |
Time of Death |
Macroscopic Observations |
5.11 |
1 Male |
Terminal Kill Day 14 |
No abnormalities detected |
2 Male |
Terminal Kill Day 14 |
No abnormalities detected |
|
3 Male |
Terminal Kill Day 14 |
No abnormalities detected |
|
4 Male |
Terminal Kill Day 14 |
No abnormalities detected |
|
5 Male |
Terminal Kill Day 14 |
No abnormalities detected |
|
6 Female |
Terminal Kill Day 14 |
No abnormalities detected |
|
7 Female |
Terminal Kill Day 14 |
No abnormalities detected |
|
8 Female |
Terminal Kill Day 14 |
No abnormalities detected |
|
9 Female |
Terminal Kill Day 14 |
No abnormalities detected |
|
10 Female |
Terminal Kill Day 14 |
No abnormalities detected |
Applicant's summary and conclusion
- Interpretation of results:
- GHS criteria not met
- Remarks:
- 4 hour LD50 is >5.11 mg/L
- Conclusions:
- The acute inhalation median lethal concentration (4 hour LC50) of FRET 13-0460 was greater than 5.11 mg/L.
- Executive summary:
FRET 13-0460 was assessed for acute toxicity via inhalation using testing guideline OECD 403. In this study, 10 rats (5 males and 5 females) were exposed to an aerosol atmosphere of the test item at 5.11 mg/L. One day after exposure, all animals exhibited hunched posture and pilo-erection, with occasional instances of sneezing. Noisy respiration was noted in one male. Animals recovered to appear normal from Days 2 to 3 post-exposure, however one male subsequently exhibited noisy respiration from Days 3 to 5 post-exposure and appeared normal from Day 6 post-exposure. All animals exhibited body weight losses or no body weight gain on the first day post-exposure. With the exception of one female from Days 1 to 3 post-exposure and three females from Days 3 to 7 post-exposure which exhibited body weight loss, body weight gains were noted for all animals during the remainder of the recovery period. No macroscopic abnormalities were detected amongst animals at necropsy. The inhalatory LD50 for the substance in male and female rats was determined to be > 5.11 mg/L.
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