Registration Dossier

Administrative data

Description of key information

Skin irritation: not irritating (OECD 439, GLP, K, Rel.1 ; OECD 404, GLP, K, Rel.1)

Eye irritation: not irritating (OECD 405, GLP, K, Rel.1)

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin irritation: in vitro / ex vivo
Remarks:
in vitro
Type of information:
experimental study
Adequacy of study:
key study
Study period:
12-17 March 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
GLP study conducted according to OECD test Guideline No. 439 with minor deviation.
Qualifier:
according to
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Deviations:
yes
Remarks:
historical data are not reported
Qualifier:
according to
Guideline:
EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
Deviations:
yes
Remarks:
historical data are not reported
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes (incl. certificate)
Remarks:
UK GLP Compliance Programme (inspected on 12-14 March 2014 / signed on 12 May 2014)
Specific details on test material used for the study:
- Purity test date: 14 January 2014
Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Justification for test system used:
Following the REACH bottom-up strategy, the EPISKIN™ Reconstructed Human Epidermis Model method was used to assess skin irritation as recommended in the OECD test guideline No. 439.
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EPISKIN™ Reconstructed Human Epidermis Model Kit
- Tissue batch number(s): 14-EKIN-008
- Production date: not reported
- Shipping date: 11 March 2014
- Delivery date: 11 March 2014
- Expiry date: 17 March 2014
- Date of initiation of testing: 12 March 2014

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

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: not reported
At the end of the exposure period, each tissue was removed from the well using forceps and rinsed using a wash bottle containing DPBS with Ca++ and Mg++. Rinsing was achieved by filling and emptying each tissue insert for approximately 40 seconds using a constant soft stream of DPBS to gently remove any residual test item.
- Observable damage in the tissue due to washing: none reported
- Modifications to validated SOP: none reported

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 0.3 mg/mL in DPBS
- Incubation time: 3 hours
- Spectrophotometer: Anthos 2001 microplate reader
- Wavelength: 562 nm (without a reference filter)
- Filter: not applicable
- Filter bandwidth: not applicable
- Linear OD range of spectrophotometer: no data reported

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: negative control OD values: 0.829, 0.830 and 0.825 (historical control not reported).
- Barrier function: IC50= 2.4 mg/mL ( ≥ 1.5 mg/mL)
- Morphology: well differenciated epidermis consisting of a basal layer, several spinous and granular layers and a thick stratum corneum
- Contamination: : absence of bacteria, fungus and mycoplasma
- Reproducibility: not reported

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
Not applicable

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

PREDICTION MODEL / DECISION CRITERIA
- The test substance is considered to be irritant to skin if the viability after the 15-min exposure period followed by the 42h post-exposure period is less or equal to 50%
- The test substance is considered to be non-irritant to skin if the viability after the 15-min exposure period followed by the 42h post-exposure period is greater than 50%
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 (26.3 μL /cm2) of the test item was applied to the epidermis surface.
- Concentration (if solution): Undiluted

VEHICLE
not applicable

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 10 μL of DPBS
- Concentration (if solution): undiluted

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 10 μL of SDS
- Concentration (if solution): 5% w/v aqueous solution



Duration of treatment / exposure:
Triplicate tissues were treated with the test item for an exposure period of 15 minutes at room temperature.
At the end of the exposure period, tissues were rinsed and incubated at 37 °C, 5% CO2 in air for 42 h.
Duration of post-treatment incubation (if applicable):
- On Day 3, at the end of the 42 h post-treatment incubation period: MTT test (MTT Loading/Formazan Extraction) was performed and tissues were incubated for 3 h at 37 °C, 5 % CO2 in air.
- On Day 6, at the end of the formazan extraction period: The optical density was measured at 562 nm (without a reference filter) using the Anthos 2001 microplate reader.
Number of replicates:
Triplicate tissues for test item, negative and positive controls
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
15 minute exposure period and 42 h post-exposure incubation period.
Value:
98.9
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:
The relative mean viability of the test item treated tissues was 98.9% after a 15 minute exposure period and 42 h post-exposure incubation period. It was considered unnecessary to perform IL-1α analysis as the results of the MTT test were unequivocal.

- OTHER EFFECTS:
- Visible damage on test system: no
- Direct-MTT reduction: no
- Colour interference with MTT: no

DEMONSTRATION OF TECHNICAL PROFICIENCY: yes

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes
- Acceptance criteria met for variability between replicate measurements: yes
- Range of historical values if different from the ones specified in the test guideline: not applicable

Table 7.3.1/1: Mean OD562Values and Percentage Viabilities for the Negative Control Item, Positive Control Item and Test Item

 

Item

OD562 of tissues

Mean OD562 of triplicate tissues

± SD of OD562

Relative individual tissue viability (%)

Relative mean viability (%)

± SD of Relative mean viability (%)

Negative Control Item

0.829

0.828

0.003

100.1

100*

0.3

0.830

100.2

0.825

99.6

Positive Control Item

0.047

0.046

0.019

5.7

5.5

2.3

0.026

3.1

0.064

7.7

Test Item

0.900

0.819

0.072

108.7

98.9

8.8

0.795

96.0

0.761

91.9

SD=        Standard deviation

*=         The mean viability of the negative control tissues is set at 100%

 

Quality Criteria

The relative mean tissue viability for the positive control treated tissues was 5.5% relative to the negative control treated tissues and the standard deviation value of the percentage viability was 2.3%. The positive control acceptance criterion was therefore satisfied.

The mean OD562 for the negative control treated tissues was 0.828 and the standard deviation value of the percentage viability was 0.3%. The negative control acceptance criterion was therefore satisfied.

The standard deviation calculated from individual percentage tissue viabilities of the three identically test item treated tissues were 8.8%. The test item acceptance criterion was therefore satisfied.

Interpretation of results:
GHS criteria not met
Conclusions:
Under the experimental conditions of this study, the test item is not classified according to Regulation (EC) No. 1272/2008 (CLP) and to the GHS.
Executive summary:

An in vitro skin irritation study was performed according to the OECD Guideline 439 and in compliance with GLP, using the EPISKINTM reconstructed human epidermis model.

 

Triplicate tissues were treated with the test item for an exposure period of 15 minutes. At the end of the exposure period each tissue was rinsed before incubating for 42 h. At the end of the post-exposure incubation period each tissue was taken for MTT-loading. The maintenance medium from beneath each tissue was transferred to pre-labeled micro tubes and stored in a freezer for possible inflammatory mediator determination. After MTT-loading a total biopsy of each epidermis was made and placed into micro tubes containing acidified isopropanol for extraction of formazan crystals out of the MTT-loaded tissues. At the end of the formazan extraction period each tube was mixed thoroughly and duplicate 200 μL samples were transferred to the appropriate wells of a pre-labeled 96-well plate. The optical density was measured at 562 nm. Data are presented in the form of percentage viability (MTT reduction in the test item treated tissues relative to negative control tissues).

 

The relative mean viability of the test item treated tissues was 98.9% after the 15 minute exposure period and 42 h post-exposure incubation period.

 

The relative mean tissue viability for the positive control treated tissues was 5.5% relative to the negative control treated tissues and the standard deviation value of the percentage viability was 2.3%. The mean OD562 for the negative control treated tissues was 0.828 and the standard deviation value of the percentage viability was 0.3%. The positive and negative controls acceptance criterion were therefore satisfied. The standard deviation calculated from individual percentage tissue viabilities of the three identically test item treated tissues were 8.8%. The test item acceptance criterion was therefore satisfied.

 

Under the experimental conditions of this study, the test item is not classified according to Regulation (EC) No. 1272/2008 (CLP) and to the GHS.

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
08-15 April 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
GLP study conducted in compliance with OECD Guideline No. 404 without any deviation.
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
Deviations:
no
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes (incl. certificate)
Remarks:
UK GLP Compliance Programme (inspected on 10 July 2012 / signed on 30 November 2012)
Specific details on test material used for the study:
- Purity test date: 14 January 2014
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories UK Ltd., Leicestershire, UK.
- Age at study initiation: 12-20 weeks
- Weight at study initiation: 2.09-2.84 kg
- Housing: Animals were individually housed in suspended cages.
- Diet: Food (2930C Teklad Global Rabbit diet supplied by Harlan Laboratories UK Ltd., Oxon, UK), ad libitum
- Water: Mains drinking water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 17-23 °C
- Humidity: 30-70 %
- Air changes: 15 changes per hour
- Photoperiod: 12 h dark / 12 h light

IN-LIFE DATES: 08-15 April 2014
Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL
- Concentration (if solution): Undiluted
- For the purpose of the study the test item was used as supplied.
Duration of treatment / exposure:
4 h
Observation period:
1, 24, 48 and 72 h and 7 days after the removal of the patch
Number of animals:
3 males
Details on study design:
PRETREATMENT
- On the day before the test each of a group of three rabbits was clipped free of fur from the dorsal/flank area using veterinary clippers. Only animals with a healthy intact epidermis by gross observation were selected for the study.

TEST SITE
- Area of exposure: On the day of the test a suitable test site was selected on the back of each rabbit.
- Type of wrap if used: Test item was applied directly to the skin under a 2.5 cm x 2.5 cm cotton gauze patch. The patch was secured in position with a strip of surgical adhesive tape.
- To prevent the animals interfering with the patches, the trunk of each rabbit was wrapped in an elasticated corset and the animals were returned to their cages for the duration of the exposure period.

REMOVAL OF TEST SUBSTANCE
- Washing: Four hours after application the corset and patches were removed from each animal and any residual test item removed by gentle swabbing with cotton wool soaked in distilled water.
- Time after start of exposure: 4 h

OBSERVATION TIME POINTS
- Immediately following removal of the patches and approximately 1, 24, 48 and 72 h later

SCORING SYSTEM:
- Method of calculation: Test sites were examined for evidence of primary irritation and scored according to the Draize scale, as described in OECD Guideline No. 404
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1.7
Max. score:
4
Reversibility:
fully reversible within: 7 days
Remarks on result:
no indication of irritation
Irritation parameter:
edema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Remarks on result:
no indication of irritation
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
4
Remarks on result:
no indication of irritation
Irritation parameter:
edema score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
4
Remarks on result:
no indication of irritation
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0.3
Max. score:
4
Reversibility:
fully reversible within: 48 h
Remarks on result:
no indication of irritation
Irritation parameter:
edema score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0.3
Max. score:
4
Reversibility:
fully reversible within: 48 h
Remarks on result:
no indication of irritation
Irritant / corrosive response data:
- Well-defined erythema and very slight edema were noted at one treated skin site (animal No. 74162) with very slight erythema and very slight edema noted at one other treated skin site (animal No. 74164) at the 24 h observation. Well-defined erythema and very slight edema persisted at one treated skin site (animal No. 74162) at the 48 h observation with very slight erythema and very slight edema noted at this treated skin site at the 72 h observation.
- One treated skin site appeared normal at the 48 h observation and one other treated skin site appeared normal at the 7-Day observation.
- No evidence of skin irritation was noted at one treated skin site during the study.
- In addition, there were no overt signs of systemic toxicity during the study.
Other effects:
None

Table 7.3.1/1: Mean irritant/corrosive response data for each animal at each observation time

Score at time point

Erythema (Animal no 1 / 2 / 3)

Max. score 4

Edema (Animal no 1 / 2 / 3)

Max. score 4

Immediately

0 / 0 / 0

0 / 0 / 0

1 h

0 / 0 / 0

0 / 0/ 0

24 h

2 / 0 / 1

1/ 0 / 1

48 h

2 / 0 / 0

1/ 0 / 0

72 h

1 / 0 / 0

1/ 0 / 0

7 days

0 / 0 / 0

0 / 0 / 0

Average 24, 48 and 72 h

1.7 / 0.0 / 0.3

1.0 / 0.0 / 0.3

 

Body weight: All animals showed expected gain in body weight during the study.

Interpretation of results:
GHS criteria not met
Conclusions:
Under the test conditions, test substance is not classified according to the Regulation (EC) No. 1272/2008 (CLP) and to the GHS.
Executive summary:

In a dermal irritation study performed according to the OECD Guideline No. 404, and in compliance with GLP, 0.5 mL of test substance was applied on the clipped skin of the dorsal flank area of 3 male New Zealand White rabbits. Test sites were covered with a semi-occlusive dressing for 4 h. Skin irritation was assessed and scored according to the Draize scale at 1, 24, 48 and 72 h and 7 days after removal of the patch.

 

Well-defined erythema and very slight edema were noted at one treated skin site with very slight erythema and very slight edema noted at one other treated skin site at the 24 h observation. Well-defined erythema and very slight edema persisted at one treated skin site at the 48 h observation with very slight erythema and very slight edema noted at this treated skin site at the 72 h observation. One treated skin site appeared normal at the 48 h observation and one other treated skin site appeared normal at the 7 days observation. No evidence of skin irritation was noted at one treated skin site during the study. There were no overt signs of systemic toxicity during the study.

 

The individual scores for each animal within 3 scoring times (24, 48 and 72 h) were 1.7 / 0.0 / 0.3 for erythema and 1.0 / 0.0 / 0.3 for oedema.

 

Under the test conditions, test substance is not classified according to the Regulation (EC) No. 1272/2008 (CLP) and to the GHS.

This study is considered as acceptable and satisfies the requirement for skin irritation endpoint.

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

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
16 June to 03 July 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
GLP study conducted in compliance with OECD Guideline No. 405 without deviation.
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes (incl. certificate)
Remarks:
UK GLP Compliance Programme (inspected on 12-14 March 2014 / signed on 12 May 2014)
Specific details on test material used for the study:
- Purity test date: 14 January 2014
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories UK Ltd., Leicestershire, UK.
- Age at study initiation: 12-20 weeks
- Weight at study initiation: 2.29-2.36 kg
- Housing: Animals were housed individually in suspended cages.
- Diet: Food (2930C Teklad Global Rabbit diet supplied by Harlan Laboratories UK Ltd., Oxon, UK), ad libitum.
- Water: Mains drinking water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 17-23 °C
- Humidity: 30-70 %
- Air changes: 15 changes/h
- Photoperiod: 12 h dark / 12 h light

IN-LIFE DATES: 16 June to 03 July 2014
Vehicle:
unchanged (no vehicle)
Controls:
other: untreated eye served as control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL
- Concentration (if solution): Undiluted
Duration of treatment / exposure:
No washing was done.
Observation period (in vivo):
1, 24, 48 and 72 h after instillation of test material
Number of animals or in vitro replicates:
3 (1 female and 2 males)
Details on study design:
PRE_TREATMENT:
Immediately before the start of the test, both eyes of the provisionally selected test rabbits were examined for evidence of ocular irritation or defect with the aid of a light source from a standard ophthalmoscope. Only animals free of ocular damage were used.

PROCEDURE:
Initially, a single rabbit was treated. A subcutaneous injection of buprenorphine 0.01 mg/kg was administered 60 minutes prior to test item application to provide a therapeutic level of systemic analgesia. Five minutes prior to test item application, a pre-dose anesthesia of ocular anesthetic (two drops of 0.5% tetracaine hydrochloride) was applied to each eye.
A volume of 0.1 mL of the test item was placed into the conjunctival sac of the right eye, formed by gently pulling the lower lid away from the eyeball. The upper and lower eyelids were held together for about one second immediately after treatment, to prevent loss of the test item, and then released.
Eight hours after test item application, a subcutaneous injection of post-dose analgesia, buprenorphine 0.01 mg/kg and meloxicam 0.5 mg/kg, was administered to provide a continued therapeutic level of systemic analgesia. The treated animal was checked for signs of pain and suffering approximately 12 hours later. No further analgesia was required.
After consideration of the ocular responses produced in the first treated animal, two additional animals were similarly treated.

REMOVAL OF TEST SUBSTANCE
- Washing: No

SCORING SYSTEM: Draize scale as described in the OECD guideline No. 405.

TOOL USED TO ASSESS SCORE: Examination of the eye was facilitated by the use of the light source from a standard ophthalmoscope.
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
4
Remarks on result:
no indication of irritation
Irritation parameter:
iris score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
fully reversible within: 24 h
Remarks on result:
no indication of irritation
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1.7
Max. score:
3
Reversibility:
fully reversible within: 7 days
Remarks on result:
no indication of irritation
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Remarks on result:
no indication of irritation
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
4
Remarks on result:
no indication of irritation
Irritation parameter:
iris score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
2
Remarks on result:
no indication of irritation
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0.7
Max. score:
3
Reversibility:
fully reversible within: 72 h
Remarks on result:
no indication of irritation
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0.3
Max. score:
4
Reversibility:
fully reversible within: 48 h
Remarks on result:
no indication of irritation
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
4
Remarks on result:
no indication of irritation
Irritation parameter:
iris score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
2
Remarks on result:
no indication of irritation
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0.7
Max. score:
3
Reversibility:
fully reversible within: 72 h
Remarks on result:
no indication of irritation
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0.3
Max. score:
4
Reversibility:
fully reversible within: 48 h
Remarks on result:
no indication of irritation
Irritant / corrosive response data:
No initial pain reaction was noted in any animal following instillation of the test item.
No corneal effects were noted during the study.
Iridial inflammation was noted in one treated eye one hour after treatment.
Moderate conjunctival irritation was noted in all treated eyes 1 h after treatment. Moderate conjunctival irritation was noted in one treated eye with minimal conjunctival irritation noted in two treated eyes at the 24 h observation. Minimal conjunctival irritation was noted in all treated eyes at the 48 h observation and persisted in one treated eye at the 72 h observation.
Two treated eyes appeared normal at the 72 h observation and one treated eye appeared normal at the 7 day observation.
Other effects:
All animals showed expected gain in body weight during the study.

Table 7.3.2/1: Eye irritation response data for each animal at each observation time

Score at time point

Cornea

Iris

(/2)

Conjunctivae

Opacity

(/4)

Area

(/4)

Redness

(/3)

Chemosis

(/4)

Discharge

(/3)

1 h

0 / 0 / 0

0 / 0 / 0

1 / 0 / 0

2 / 2 / 2

1 / 1 / 1

2 / 1 / 1

24 h

0 / 0 / 0

0 / 0 / 0

0 / 0 / 0

2 / 1 / 1

1 / 1 / 1

1 / 0 / 0

48 h

0 / 0 / 0

0 / 0 / 0

0 / 0 / 0

2 / 1 / 1

1 / 0 / 0

0 / 0 / 0

72 h

0 / 0 / 0

0 / 0 / 0

0 / 0 / 0

1 / 0 / 0

1 / 0 / 0

0 / 0 / 0

7 days

0 / - / -

0 / - / -

0 / - / -

0 / - / -

0 / - / -

0 / - / -

Average 24, 48 and 72 h

0.0 / 0.0 / 0.0

0.0 / 0.0 / 0.0

0.0 / 0.0 / 0.0

1.7 / 0.7 / 0.7

1.0 / 0.3 / 0.3

0.3 / 0.0 / 0.0

Reversibility

 -

 -

Completely

reversible

Completely

reversible

Completely reversible

Completely reversible

Average time (unit) for reversion

-

-

24 h

7 days

7 days

48 h

Interpretation of results:
GHS criteria not met
Conclusions:
Under the test conditions, the test substance is not classified as irritating to eyes according to the criteria of the Regulation (EC) No. 1272/2008 (CLP) and to the GHS.
Executive summary:

In an eye irritation study performed according to the OECD Guideline No. 405, and in compliance with GLP, 0.1 mL of undiluted test item was instilled into the right eye of 3 New Zealand White rabbits (1 female and 2 males). The upper and lower eyelids were held together for about one second immediately after application, to prevent loss of the test material, and then released. The left eye remained untreated and served as control. The eyes were examined and the changes were observed at 1, 24, 48, 72 h and 7 days after treatment and graded according to the Draize method.

No initial pain reaction was noted in any animal following instillation of the test item. No corneal effects were noted during the study. Iridial inflammation was noted in one treated eye one hour after treatment. Moderate conjunctival irritation was noted in all treated eyes 1 h after treatment. Moderate conjunctival irritation was noted in one treated eye with minimal conjunctival irritation noted in two treated eyes at the 24 h observation. Minimal conjunctival irritation was noted in all treated eyes at the 48 h observation and persisted in one treated eye at the 72 h observation. Two treated eyes appeared normal at the 72 h observation and one treated eye appeared normal at the 7 day observation.

Mean individual scores at 24, 48 and 72 h after exposure for the 3 animals were 0.0 / 0.0 / 0.0 for cornea score; 0.0 / 0.0 / 0.0 for iris score; 1.7 / 0.7 / 0.7 for conjunctivae score and 1.0 / 0.3 / 0.3 for chemosis score.

Under the test conditions, the test substance is not classified as irritating to eyes according to the criteria of the Regulation (EC) No. 1272/2008 (CLP) and to the GHS.

This study is considered as acceptable and satisfies the requirement for eye irritation endpoint.

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

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation:

Table 7.3/1: Summary of skin irritation tests

 

Test n°

Test / Guideline

Reliability

Focus

Results

1

 

Harlan, 2014

In vitro skin irritation

(OECD 439)

K, rel. 1

Skin irritation

non irritant

2

 

Harlan, 2014

Acute dermal irritation/corrosion (OECD 404)

K, rel. 1

Skin irritation

 non irritant

 

Two key studies were identified (Test No.1 and 2) with both negative result.

- In vitro EPISKIN (Test No. 1):

The purpose of this in vitro test was to evaluate the skin irritation potential of the test substance using the EPISKIN reconstructed human epidermis model after a treatment period of 15 minutes followed by a post-exposure incubation period of 42 hours. This test was designed to be compatible with the OECD Guideline No. 439 and was performed in compliance with GLP. The quality criteria required for acceptance of results in the test were satisfied. The relative mean viability of the test item treated tissues was 98.9%, after the 15-minute exposure period and 42 hours post-exposure incubation period. With a tissue viability > 50%, the test substance was considered to be non-irritant to skin.

- In vivo OECD 404 (Test No. 2):

Although not needed under the REACH Regulation, the in vivo study was requested for China notification purpose.

This dermal irritation study was performed according to the OECD Guideline No. 404, and in compliance with GLP. A single 4-Hour, semi-occluded application of the test item to the intact skin of three rabbits produced very slight to well-defined erythema and very slight edema. No evidence of skin irritation was noted at one treated skin site. One treated skin site appeared normal at the 48-Hour observation and one other treated skin site appeared normal at the 7-Day observation. The individual scores for each animal within 3 scoring times (24, 48 and 72 h) were 1.7/0.0/0.3 for erythema and 1.0/0.0/0.3 for oedema. The effects observed were all reversible within 7 days.

Conclusion: Both studies showed the same result. The test substance is considered to be non-irritant for the skin.

 

Eye irritation:

A key study was identified (Harlan, 2015). In this eye irritation study performed according to the OECD guideline 405, and in compliance with GLP, 0.1 mL of undiluted test material was instilled into one eye of 3 female New Zealand White rabbits. The other eye remained untreated and served as control. The upper and lower eyelids were held together for about one second immediately after instillation. The eyes were examined and the changes were observed at 1, 24, 48 and 72 h after treatment and graded according to the Draize method.

The calculated mean score for each animal within 3 scoring times (24, 48 and 72 h) were 0.0/0.0/0.0 for the corneal opacity and iridial inflammation, 1.7/0.7/0.7 for conjunctival redness and 1.0/0.3/0.3 for chemosis. The effects observed were all reversible within 7 days. The substance is therefore not classified for eye irritation.

Justification for classification or non-classification

Harmonized classification:

The substance has no harmonized classification according to the Regulation (EC) No. 1272/2008.

 

Self classification:

Based on the available data, no additional self-classification is proposed regarding both skin and eye irritation according to the Regulation (EC) No. 1272/2008 (CLP) and according to the GHS.

 

No data was available regarding respiratory irritation. However, the substance not being classified for skin and eye irritation, no classification is expected for respiratory irritation .