Registration Dossier

Administrative data

Description of key information

Using in vitro assays, the substance was found not corrosive to skin in an OECD TG 431 and not irritating in the OECD TG 439/EU method B.46.

The substance was found not severely damaging to eyes in the in vitro BCOP assay (OECD TG 437), and an in vivo study according to OECD TG 405 confirmed the substance was not irritating to eyes.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin irritation / corrosion
Remarks:
in vitro
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)
Deviations:
no
GLP compliance:
yes
Species:
other: in vitro method
Strain:
other: in vitro method
Details on test animals or test system and environmental conditions:
TEST SYSTEM: Reconstructed human epidermal tissue (EpiDerm EPI-200 model).
Type of coverage:
other: not applicable: in vitro method
Preparation of test site:
other: not applicable: in vitro method
Vehicle:
unchanged (no vehicle)
Controls:
other: not applicable: in vitro method
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 50 µL
Duration of treatment / exposure:
3 minute and 1 hour exposures were conducted.
Observation period:
Following the exposure periods, the tissues were washed with phosphate buffered saline (PBS). The DMEM medium was replaced with 300 µL MTT medium and then all tissues were again incubated for 3 hours at 37 C in 5% CO2. The tissues were then washed with PBS and formazan was extracted with 2 mL isopropanol.
Number of animals:
None: in vitro method
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): Following the exposure periods, the tissues were washed with phosphate buffered saline (PBS).
- Time after start of exposure: 3 minutes or 1 hour.

SCORING SYSTEM: The DMEM medium was replaced with 300 µL MTT medium and then all tissues were again incubated for 3 hours at 37°C in 5% CO2. The tissues were then washed with PBS and formazan was extracted with 2 mL isopropanol. The amount of extracted formazan was determined spectrophotometrically at 540 nm in triplicate. Cell viability was then calculated as a percentage of the mean of the control tissues, and skin corrosion potential calculated from the optical density at 540 nm (OD540) measurements.
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
3 minute and 1 hour exposure
Value:
ca. 70 - 89
Vehicle controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: Not corrosive in vitro
Other effects / acceptance of results:
The mean tissue viability of the test article was 70% of the negative control after a 3 minute exposure and 89% after 1 hour exposure. The controls performed as expected indicating that the test was valid.
Interpretation of results:
other: Not corrosive
Conclusions:
Based on the results of the study, the test article is not corrosive in the in vitro skin corrosion test model.
Executive summary:

The skin corrosion potential of the test article (clear colourless liquid, Purity: 99.64%, Lot: 12912) was evaluated in an in vitro human skin model test. This study was performed in compliance with OECD GLP (1997). The study design was based on OECD Guideline No. 431 (2013) and EC 440/2008 (2008).

 

Human-derived keratinocytes were cultured to form a multi-layered, highly differentiated model of the human epidermis (EpiDerm EPI-200 model). The tissues were kept refrigerated until the day of use, transferred to 6 well plates containing 0.9 mL DMEM medium, and then incubated for 2 hours at 37°C in 5% CO2. The test article (50 µL) was applied to 2 tissues for a 3 minute exposure and to 2 tissues for 1 hour exposure. The negative control (Milli-Q water) and the positive control (8N Potassium hydroxide) were applied (50 µL of each) in the same manner. Following the exposure periods, the tissues were washed with phosphate buffered saline (PBS). The DMEM medium was replaced with 300 µL MTT medium and then all tissues were again incubated for 3 hours at 37°C in 5% CO2. The tissues were then washed with PBS and formazan was extracted with 2 mL isopropanol. The amount of extracted formazan was determined spectrophotometrically at 540 nm in triplicate. Cell viability was then calculated as a percentage of the mean of the control tissues, and skin corrosion potential calculated from the optical density at 540 nm (OD540) measurements.

 

The mean tissue viability of the test article was 70% of the negative control after a 3 minute exposure and 89% after 1 hour exposure. The controls performed as expected indicating that the test was valid.

 

Based on the results of the study, the test article is not corrosive in the in vitro skin corrosion test model.

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
27-JUN-2017 to 07-AUG-2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: BLT 03-2017
- Expiration date of the lot/batch: March 2022
- Purity test date: 10 March 2017

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Controlled room temperature (15-25 ºC, below 70 RH%), protected from light

Test system:
human skin model
Remarks:
reconstructed human epidermis model
Source species:
human
Cell type:
non-transformed keratinocytes
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EPISKIN model supplied by SkinEthic, France, three-dimensional human epidermis model
- Tissue batch number(s): 17-EKIN-026
- Production date: not reported, Quality control 27 June 2017
- Shipping date: not reported
- Delivery date: not reported
- Expiry date: 03 July 2017
- Date of initiation of testing: 29 June 2017

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 27.2 - 28°C (room temperature) for 15 min
- Temperature of post-treatment incubation (if applicable): 37°C

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: Thorough PBS washes to remove any remaining material (no details)
- Observable damage in the tissue due to washing: No
- Modifications to validated SOP: None

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 0.3 mg/ml MTT per well in 12-well plates
- Incubation time: 3 hours (+/- 5 min)
- Spectrophotometer: Thermo Fisher Scientific
- Wavelength: 570 nm

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability:
- Barrier function:
- Morphology:
- Contamination:
- Reproducibility:

NUMBER OF REPLICATE TISSUES: 3

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE : No MTT interference detected

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: 1 assay

PREDICTION MODEL / DECISION CRITERIA (According to OECD439)
- The test substance is considered to be irritating to skin (UN GHS category 2) if the viability after 15 minutes exposure is less or equal to 50% compared to the negative control
- The test substance is considered to be non-irritant to skin if the viability after 15 minutes exposure is greater than to 50% compared to the negative control
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 10 µl
- Concentration : undiluted

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 50 µl
- Concentration (if solution): as supplied

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 50 µl
- Concentration (if solution): 5% (w/v) SDS solution
Duration of treatment / exposure:
15 minutes exposure
Duration of post-treatment incubation (if applicable):
After rinsing the units were placed into the plate wells with fresh pre-warmed Maintenance Medium (2 mL/well) below them and then incubated for 42 hours (± 1h) at 37°C in an incubator with 5% CO2, in a >95% humidified atmosphere.
Number of replicates:
6 wells for blank OD values
3 wells for the test item samples
3 wells for negative control samples
3 wells for positive control samples
Irritation / corrosion parameter:
% tissue viability
Value:
ca. 89.2
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Other effects / acceptance of results:
- OTHER EFFECTS:
- Visible damage on test system: no visible damage
- Direct-MTT reduction: No direct reaction of the test item with MTT was detected in the pre-test check following a 3-hour incubation of the test item (10 µl) with 2 ml MTT working solution.
- Colour interference: As the test item was coloured two additional test item-treated tissues were used for the non-specific OD evaluation. The mean optical density (measured at 570 nm) of tissues were 0.012, Non Specific Colour % was calculated as 1.7%. This value was below 5%, therefore additional data calculation was not necessary.


ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: The mean corrected OD value of the three negative control tissues was in the recommended range (0.706), with SD of the viability % of 0.8. (criteria: between 0.6 and 1.5, and SD of the % viability should be - Acceptance criteria met for positive control: The positive control treated tissues showed 4.7% viability demonstrating the proper performance of the assay (SD: 1.1). (Criteria for acceptablr viability range for positive control 0-40% with SD - Acceptance criteria met for variability between replicate measurements: The standard deviation of viability values of the three test item-treated tissue samples in the MTT assay was 5.1 (Criteria: SD - Range of historical values if different from the ones specified in the test guideline: One individual opacity density data (0.026) in this study was slightly lower than the lower limit of the historical control range (0.032). This fact had no impact on the results or integrity of the study since the positive control material showed severe effect.
Interpretation of results:
GHS criteria not met
Conclusions:
Following exposure with 1,4-Diiodoperfluorobutane, the mean cell viability was 89.2% compared to the negative control. This is above the threshold of 50%, therefore the test item was considered as being non-irritant to skin. The experiment met the validity criteria, therefore the study was considered to be valid.
Executive summary:

The skin irritation potential of 1,4-Diiodoperfluorobutane was assessedin vitroaccording to the OECD No. 439 guideline using a reconstructed human epidermis model by measuring its cytotoxic effect as reflected in the MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay.

Disks of EPISKINTM(SM) (three units) were treated with the test item and incubated for 15 minutes at room temperature. Exposure of the test item was terminated by rinsing with Phosphate Buffered Saline (PBS). The epidermis units were then incubated at 37°C for 42 hours in an incubator with 5% CO2 in a >95% humidified atmosphere. The viability of each disk was assessed by incubating the tissues for 3 hours with MTT solution at 37°C in an incubator with 5% CO2 in a >95% humidified atmosphere protected from light. The precipitated formazan crystals were then extracted using acidified isopropanol and quantified spectrophotometrically.

PBS and 5% (w/v) Sodium Dodecyl Sulphate (SDS) solution treated epidermis were used as negative and positive controls, respectively (three units / control). Two additional disks were used to provide an estimate of colour contribution (NSCliving) from the test item. For each treated tissue, the viability was expressed as a % relative to the negative control. If the mean relative viability after 15 minutes exposure and 42

hours post incubation is less or equal (≤) to 50% of the negative control, the testitem is considered to be irritant to skin.

Following exposure with 1,4-Diiodoperfluorobutane, the mean cell viability was 89.2% compared to the negative control. This is above the threshold of 50%, therefore the test item was considered as being non-irritant to skin. The experiment met the validity criteria, therefore the study was considered to be valid.

In conclusion, in thisin vitroEPISKINTM (SM) model test with 1,4-Diiodoperfluorobutane, the results indicate that the test item is non-irritant to skin.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Referenceopen allclose all

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)
Deviations:
no
GLP compliance:
yes
Species:
other: in vitro
Strain:
other: in vitro
Details on test animals or tissues and environmental conditions:
Not applicable: in vitro
Vehicle:
unchanged (no vehicle)
Controls:
other: not applicable: in vitro method
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 750 µL
Duration of treatment / exposure:
Corneas were incubated with the test article or appropriate control for 10 minutes at 32 degrees C.
Duration of post- treatment incubation (in vitro):
After rinsing with Minimal Essential Medium (MEM), the corneas were incubated for 120 minutes in Eagle's Minimum Essential Medium. After the completion of the incubation period, opacity was evaluated using an opacitometer. The cell culture medium was then replaced with Na-fluorescein medium and incubated for approximately 90 minutes at 32 C. Following the 90 minute Na-fluorescein exposure, permeability was measured spectrophotometrically.
Number of animals or in vitro replicates:
0, in vitro method
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): The corneas were rinsed with MEM after a 10 minute exposure to the test article or appropriate control.
- Time after start of exposure: 10 minutes

SCORING SYSTEM: Corneal opacity and permeability were measured and used to calculate an in vitro irritancy score (IVIS)

TOOL USED TO ASSESS SCORE: TECAN Infinite M200 Pro Plate Reader
Irritation parameter:
in vitro irritation score
Remarks:
IVIS
Run / experiment:
Score following a 10-minute exposure
Value:
3.9
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
positive indication of irritation
Remarks:
measured as mean opacity value + (15 x mean OD450 value)
Other effects / acceptance of results:
The mean in vitro Irritancy Score (IVIS) score following a 10 minutes exposure was 3.9.
Interpretation of results:
other: Does not cause severe ocular damage
Conclusions:
Based on the results of the test (IVIS=3.9), the test substance is not Category 1, and it is not “no category.” No conclusion can be reached on ocular irritation potential.
Executive summary:

The corneal irritation and damage potential of the test article (clear and colourless liquid, Purity 99.64%, Lot 12912) was tested in the Bovine Corneal Opacity and Permeability test (BCOP test). The study was performed in compliance with OECD GLP (98) 17 (1997). The test method was based on OECD no. 437 (2009), EC No. 440/2008 B. 47 (2010), OTWG- ICCVAM-NICEATM (2006), INVITTOX 127 (2006), and Gautheron, P. et al. 18: 442-449 (1992).

 

Corneas were prepared in cell culture medium and incubated at 32°C for at least 1 hour prior to exposure. Corneas (3/group) were treated with 0.75 mL of undiluted test material and incubated in a horizontal position for 10 minutes at 32°C. A positive control (10% w/v Benzalkonium Chloride) and a negative control (physiological saline) were tested in parallel with the test material. At the end of the exposure period, the corneas were rinsed and incubated in fresh cell culture medium for 120 ± 10 minutes at 32 C. Opacity was evaluated using an opacitometer after the 2 hour post-exposure incubation. Following opacity readings, the cell culture medium was replaced with Na-fluorescein medium and incubated for approximately 90 minutes at 32°C. Following the 90 minute Na-fluorescein exposure, permeability was measured. The mean in vitro irritation score (IVIS) was calculated and the scores were classified according to protocol defined categories.

 

For the test article, the mean IVIS = 3.9 after 10 minutes. The mean opacity score was 4 (range 3 to 5) and the mean permeability score was 0.-0.004 (range -0.012 to 0.001). Controls performed as expected. No pH effect of the test substance was observed on the rinsing medium. 

 

Based on the results of the test (IVIS=3.9), the test substance is not Category 1, and it is not “no category.”

Endpoint:
eye irritation: in vivo
Remarks:
This in vivo study was conducted to meet regulatory requirements outside Europe
Type of information:
experimental study
Adequacy of study:
key study
Study period:
31-AUG-2017 to 01-DEC-2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
2nd October 2012
Deviations:
yes
Remarks:
some deviations of temperature and humidity values in the room, considered to have no impact on the outcome of the study
Qualifier:
according to guideline
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Version / remarks:
L 152 2004
Deviations:
yes
Remarks:
some deviations of temperature and humidity values in the room, considered to have no impact on the outcome of the study
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: BLT 03-2017
- Expiration date of the lot/batch: March 2022
- Purity test date: 10 March 2017

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material:
Controlled room temperature (15-25 ºC, below 70 RH%), protected from light

Species:
rabbit
Strain:
New Zealand White
Remarks:
Crl:KBL (NZW)
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories
- Age at study initiation: 13 weeks old
- Weight at study initiation: 2964 g – 3331 g (on day of treatment)
- Housing: individually housed in AAALAC approved metal wire rabbit cages. Cages were of an open wire structure and cages were placed together to allow some social interaction with rabbit(s) in adjoining cages.
- Diet : ad libitum
- Water : ad libitum
- Acclimation period: At least 33 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.3 – 23.3°C
- Humidity (%): 40 – 73%
- Air changes (per hr): 15-20 air exchanges/hour
- Photoperiod (hrs dark / hrs light): 12 hours daily, from 6.00 a.m. to 6.00 p.m.

IN-LIFE DATES: From: 12-SEP-2017 To: 15 or 17-SEP-2017
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume): 0.1 ml, undiluted
Duration of treatment / exposure:
The test item was placed in the conjunctival sac of the left eye of each animal. No test item remained in the eye sac of any animal at 1 hour after the application (time of 1st observation), therefore no washing was performed.
Observation period (in vivo):
The eyes were examined at 1, 24, 48, 72 hours after treatment;
As no clinical signs were observed, the experiment was terminated after 72 hours observation of the third rabbit
Number of animals or in vitro replicates:
3 males
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): No

SCORING SYSTEM: Draize scale

TOOL USED TO ASSESS SCORE: none
Irritation parameter:
cornea opacity score
Remarks:
individual mean score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
cornea opacity score
Remarks:
individual mean score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
cornea opacity score
Remarks:
individual mean score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
iris score
Remarks:
individual mean score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
2
Irritation parameter:
iris score
Remarks:
individual mean score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
2
Irritation parameter:
iris score
Remarks:
individual mean score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
2
Irritation parameter:
conjunctivae score
Remarks:
individual mean score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
3
Irritation parameter:
conjunctivae score
Remarks:
individual mean score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
3
Irritation parameter:
conjunctivae score
Remarks:
individual mean score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
3
Irritation parameter:
chemosis score
Remarks:
individual mean score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
chemosis score
Remarks:
individual mean score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
chemosis score
Remarks:
individual mean score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritant / corrosive response data:
Only transient conjunctival reactions (redness, chemosis and discharge graded 1) were observed at the 1-hour time point only in all 3 rabbits.
Other effects:
none reported
Interpretation of results:
GHS criteria not met
Conclusions:
The test item 1,4-Diiodoperfluorobutane, applied to rabbit eye mucosa, caused conjunctival effects at 1 hour after the treatment which were fully reversible within 24 hours. There were no corneal effects observed in the study.
According to Regulation (EC) No 1272/2008, 1,4-Diiodoperfluorobutane does not require classification as an eye irritant.
Executive summary:

An acute eye irritation study of the test item 1,4-Diiodoperfluorobutane was performed in three New Zealand White rabbits. The irritation effects of the test item were evaluated according to the Draize method (OECD No.: 405, 2012). Rabbits were treated with analgesic and anaesthetic as per the regulatory guideline.

The test item was placed into the conjunctival sac of the left eye of each animal. The untreated right eye served as control. A single amount of 0.1 ml of the test item was administered to the left eye of each animal. The eyes were examined at 1, 24, 48 and 72 hours after application.

 

No Initial Pain Reaction/Pain reaction (IPR/PR) was observed. In all three animals, at 1 hour after score 1 effects were observed for conjunctival redness, chemosis and discharge but no corneal or iris effects. No test item remained in the eye sac.

At 24, 48 and 72 hours after the application, no conjunctival or corneal effects were observed in any of the three animals. The individual mean scores were 0.

No clinical signs were observed throughout the observation period.

During the experiment, the control eye of each animal was symptom-free. Washing of the treated eyes was not necessary during the study.

As no clinical signs were observed, the experiment was terminated after 72 hours observation of the third rabbit.

The general state and behaviour of animals were normal throughout the study period. No mortality occurred during the study. The bodyweights of all rabbits were considered to be within the normal range of variability.

The test item 1,4-Diiodoperfluorobutane, applied to rabbit eye mucosa, caused conjunctival effects at 1 hour after the treatment which were fully reversible within 24 hours. There were no corneal effects observed in the study.

According to Regulation (EC) No 1272/2008, 1,4-Diiodoperfluorobutane does not require classification as an eye irritant.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Additional information

Justification for classification or non-classification

Based on the Integrated Approach to Testing and Assessment (IATA) guidance document and available experimental data, according to the CLP Regulation (EC) No 1272/2008, 1,4-Diiodoperfluorobutane does not require classification as a skin or eye irritant.