Registration Dossier

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Administrative data

Description of key information

Skin irritation: non-irritating, OECD TG 404, 2014

Eye irritation: non-irritating, OECD TG 405, 2014

Eye irritation: non-irritating, OECD TG 437 – BCOP Method, 2014

Respiratory irritation: non-irritating at 5.0 mg/L mist, OECD TG 436 (acute toxicity: inhalation), 2013

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study performed under GLP. All relevant validity criteria were met.
Qualifier:
according to guideline
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
inspected: July 2012; signature: November 2012
Species:
rabbit
Strain:
New Zealand White
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: recognised animal supplier
- Age at study initiation: 12 - 20 weeks
- Weight at study initiation: 2.37 - 2.75 kg
- Housing: Individually housed in suspended cages.
- Diet: certified rabbit diet ad libitum
- Water: mains drinking water ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 - 23
- Humidity (%): 30 - 70
- Air changes (per hr): 15
- Photoperiod: 12 hours light / 12 hours dark
Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5mL
- Concentration (if solution): Test material was used as supplied.
Duration of treatment / exposure:
4 hours
Observation period:
72 hours
Number of animals:
3
Details on study design:
TEST SITE
- Area of exposure: dorsal
- Type of wrap if used: semi-occlusive (cotton gauze patch secured with surgical adhesive tape)

REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes, only within sentinel rabbit used in preliminary test. Three patches applied. One patch was removed at each of three time points: 3 minutes, 1 hour and 4 hours after application. Any residual test item was removed by gentle swabbing with cotton wool soaked in distilled water.
In the definitive test for skin irritation (after consideration of the sentinel skin reactions) with two additional rabbits; no washing/swabbing of the test site was reported after 4 hours application and removal of the patch.
- Time after start of exposure: 4 hours

SCORING SYSTEM:
Erythema and Eschar Formation
No erythema _________________________________________________________________________0
Very slight erythema (barely perceptible) ________________________________________________1
Well-defined erythema ________________________________________________________________2
Moderate to severe erythema __________________________________________________________3
Severe erythema (beet redness) to slight eschar formation (injuries in depth) ________________4

Oedema Formation
No oedema __________________________________________________________________________0
Very slight oedema (barely perceptible) _________________________________________________1
Slight oedema (edges of area well-defined by definite raising) _____________________________2
Moderate oedema (raised approximately 1 millimetre) ____________________________________3
Severe oedema (raised more than 1 millimetre and extending beyond the area of exposure) ___4
Any other skin reactions and clinical signs of toxicity, if present, were also recorded.
Irritation parameter:
erythema score
Basis:
mean
Remarks:
(n=3)
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: no effects observed
Remarks on result:
no indication of irritation
Irritation parameter:
edema score
Basis:
mean
Remarks:
(n=3)
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: no effects observed
Remarks on result:
no indication of irritation
Irritation parameter:
erythema score
Basis:
mean
Remarks:
(n=3)
Time point:
other: 24h
Score:
0
Max. score:
4
Reversibility:
other: no effects observed
Irritation parameter:
erythema score
Basis:
mean
Remarks:
(n=3)
Time point:
other: 48h
Score:
0
Max. score:
4
Reversibility:
other: no effects observed
Irritation parameter:
erythema score
Basis:
mean
Remarks:
(n=3)
Time point:
other: 72h
Score:
0
Max. score:
4
Reversibility:
other: no effects observed
Irritation parameter:
edema score
Basis:
mean
Remarks:
(n=3)
Time point:
other: 24h
Score:
0
Max. score:
4
Reversibility:
other: no effects observed
Irritation parameter:
edema score
Basis:
mean
Remarks:
(n=3)
Time point:
other: 48h
Score:
0
Max. score:
4
Reversibility:
other: no effects observed
Irritation parameter:
edema score
Basis:
mean
Remarks:
(n=3)
Time point:
other: 72h
Score:
0
Max. score:
4
Reversibility:
other: no effects observed
Irritant / corrosive response data:
- Erythema: No evidence of skin irritation was noted during the study.
- Edema: No evidence of skin irritation was noted during the study.
- Reversibility of effects: Not applicable.
Other effects:
- Other adverse local effects: None
- Other adverse systemic effects: No signs of systemic toxicity were noted during the observation period. All animals showed expected gain in body weight during the study.

Table 1. Individual skin reactions

Skin Reaction

Observation Time

Individual Scores

 

 

1 (male)

2 (male)

3 (male)

Erythema/Eschar formation

1 hour

0

0

0

 

24 hours

0

0

0

 

48 hours

0

0

0

 

72 hours

0

0

0

 

 

 

 

 

Edema formation

1 hour

0

0

0

 

24 hours

0

0

0

 

48 hours

0

0

0

 

72 hours

0

0

0

 

 

 

 

 

 

Mean scores per organism at 24, 48 and 72h:

Erythemea/Escar Formation:

1: total = 0; mean score = 0

2: total = 0; mean score = 0

3: total = 0; mean score = 0

Edema Formation:

1: total = 0; mean score = 0

2. total = 0; mean score = 0

3. total = 0; mean score = 0

Interpretation of results:
GHS criteria not met
Remarks:
EU criteria not met
Conclusions:
Under the conditions of this study the substance is not irritating to the skin.
Executive summary:

The study was performed to OECD TG 404 and EU Method B.4 in accordance with GLP to assess the primary skin irritancy potential of the test item in New Zealand White rabbits to assess the irritancy potential of the test item following single, 3-Minute, 1 and 4-Hour, semi-occluded applications to the intact rabbit skin. 0.5 ml of the test material was introduced under a 2.5 cm x 2.5 cm cotton gauze patch and placed in position on the clipped skin. The patch was secured in position with a strip of surgical adhesive tape. 3-Minute and 1-Hour semi-occluded applications of the test item to the intact skin of one rabbit produced no evidence of skin irritation. After 4 hours of exposure to the test substance, the patches were removed and individual dose sites were scored according to the Draize scoring system at approximately 1, 24, 48, and 72 hours. A single 4-Hour, semi occluded application of the test item to the intact skin of three rabbits produced no evidence of skin irritation. Mean scores for following grading at 24, 48 and 72h were zero in both scoring criteria and there was no other signs or systemic toxicity noted during the study. Under the conditions of the study, the substance cannot be considered a skin irritant.

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

Eye irritation

Link to relevant study records

Referenceopen allclose all

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study performed under GLP. All relevant validity criteria were met.
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
inspected: July 2012; signature: November 2012
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Recognised animal supplier
- Age at study initiation: 12-20 weeks old
- Weight at study initiation: 2.38 - 2.49 kg
- Housing: individually housed in suspended metal cages
- Diet: certified rabbit food ad libitum
- Water: mains water ad libitum
- Acclimation period: at least 5 days

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

Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL
- Concentration (if solution): undiluted

Duration of treatment / exposure:
A volume of 0.1 ml of the test material, 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 material, and then released. The left eye remained untreated and was used for control purposes.
Observation period (in vivo):
The eyes of each animal were examined approximately 1, 24, 48 and 72 hours after instillation of the test substance.
Number of animals or in vitro replicates:
3 males
Details on study design:
The study was performed in a stepwise manner and was started by treatment of a single rabbit. The two other animals were treated in a similar manner after considering the degree of eye irritation observed in the first animal.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): none

SCORING SYSTEM:
The irritation was assessed according to the following numerical scoring system. At each observation, the highest scores given were recorded:

CORNEAL IRRITATION
Opacity: degree of density (area most dense taken for reading)
No ulceration or opacity (may include slight dulling of normal luster)_____0
Scattered or diffuse areas of opacity, details of iris clearly visible______1
Easily discernible translucent area, details of iris slightly obscured______2
Nacreous area, no details of iris visible, size of pupil barely discernible__3
Opaque cornea, iris not discernible through the opacity______________4

Area of cornea involved:
No ulceration or opacity________________________________________0
One quarter or less but not zero_________________________________1
Greater than one quarter, but less than half________________________2
Greater than half, but less than three quarters______________________3
Greater than three quarters, up to whole area______________________4

IRIS
Normal______________________________________________________0
Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia,
or injection, any of these or combination thereof, iris still reacting to light
(sluggish reaction is positive)____________________________________1
No reaction to light, hemorrhage, gross destruction (any or all of these)___2

CONJUNCTIVAL IRRITATION
Redness (refers to palpebrae and sclera, excluding cornea and iris):
Blood vessels normal__________________________________________0
Some blood vessels definitely hyperaemic (injected)__________________1
Diffuse, crimson color, individual vessels not easily discernible_________2
Diffuse beefy red_____________________________________________3

Chemosis (refers to lids and/or nictitating membranes):
No swelling__________________________________________________0
Any swelling above normal (includes nictitating membranes)___________1
Obvious swelling with partial eversion of the lids____________________2
Swelling with lids about half closed_______________________________3
Swelling with lids more than half closed ___________________________4

Discharge:
No discharge (may include small amounts observed in inner canthus of normal animals)_____0
Any amount different from normal and/or lacrimation_________________________________1
Discharge with moistening of the lids and hairs just adjacent to lids_____________________2
Discharge with moistening of the lids and hairs (considerable area around the eyes)_______3

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:
mean
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
iris score
Basis:
mean
Time point:
24/48/72 h
Score:
0
Max. score:
2
Irritation parameter:
conjunctivae score
Basis:
mean
Time point:
24/48/72 h
Score:
0.8
Max. score:
3
Reversibility:
fully reversible within: 72h
Irritation parameter:
chemosis score
Basis:
mean
Time point:
24/48/72 h
Score:
0.6
Max. score:
4
Reversibility:
fully reversible within: 72h
Irritation parameter:
cornea opacity score
Basis:
animal #1
Remarks:
mean
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
iris score
Basis:
animal #1
Remarks:
mean
Time point:
24/48/72 h
Score:
0
Max. score:
2
Irritation parameter:
conjunctivae score
Basis:
animal #1
Remarks:
mean
Time point:
24/48/72 h
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 72h
Irritation parameter:
chemosis score
Basis:
animal #1
Remarks:
mean
Time point:
24/48/72 h
Score:
0.3
Max. score:
4
Reversibility:
fully reversible within: 72h
Irritation parameter:
cornea opacity score
Basis:
animal #2
Remarks:
mean
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
iris score
Basis:
animal #2
Remarks:
mean
Time point:
24/48/72 h
Score:
0
Max. score:
2
Irritation parameter:
conjunctivae score
Basis:
animal #2
Remarks:
mean
Time point:
24/48/72 h
Score:
0.7
Max. score:
3
Reversibility:
fully reversible within: 72h
Irritation parameter:
chemosis score
Basis:
animal #2
Remarks:
mean
Time point:
24/48/72 h
Score:
0.7
Max. score:
4
Reversibility:
fully reversible within: 72h
Irritation parameter:
cornea opacity score
Basis:
animal #3
Remarks:
mean
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
iris score
Basis:
animal #3
Remarks:
mean
Time point:
24/48/72 h
Score:
0
Max. score:
2
Irritation parameter:
conjunctivae score
Basis:
animal #3
Remarks:
mean
Time point:
24/48/72 h
Score:
0.7
Max. score:
3
Reversibility:
fully reversible within: 72h
Irritation parameter:
chemosis score
Basis:
animal #3
Remarks:
mean
Time point:
24/48/72 h
Score:
0.7
Max. score:
4
Reversibility:
fully reversible within: 72h
Irritant / corrosive response data:
No corneal effects were noted. Iridial inflammation was noted in one treated eye one hour after treatment. Moderate conjunctival irritation was noted in all treated eyes one hour after treatment. Moderate conjunctival irritation was noted in one treated eye with minimal conjunctival irritation noted in two treated eyes at the 24-Hour observation. Minimal conjunctival irritation was noted in all treated eyes at the 48-Hour observation. All treated eyes appeared normal at the 72-Hour observation.

Table 1. Individual scores and mean scores for 24, 48 and 72 hours

Organism number and sex

Time after treatment

Corneal Opacity

Iridial Inflammation

Conjunctival Redness

Conjunctival Chemosis

1 (male)

1h

0

0

2

2

 

24 h

0

0

2

1

 

48 h

0

0

1

0

 

72 h

0

0

0

0

Mean (24 – 72 h)

 

0.00

0.00

1.0

0.3

 

 

 

 

 

 

2 (male)

1h

0

0

2

1

 

24 h

0

0

1

1

 

48 h

0

0

1

1

 

72 h

0

0

0

0

Mean (24 – 72 h)

 

0.00

0.00

0.7

0.7

 

 

 

 

 

 

3 (male)

1h

0

1

2

2

 

24 h

0

0

1

1

 

48 h

0

0

1

1

 

72 h

0

0

0

0

Mean (24 – 72 h)

 

0.00

0.00

0.7

0.7

 

Interpretation of results:
GHS criteria not met
Remarks:
EU criteria not met
Conclusions:
Under the conditions of this study the substance is not irritating to the eye.
Executive summary:

The study was performed to OECD TG 405 and EU Method B.5 under GLP to assess the irritancy potential of the test material to the eye following a single application in the New Zealand White rabbit. A volume of 0.1 ml of the test material was placed into the conjunctival sac of one eye of 3 animals. The other eye remained untreated and was used for control purposes. Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment. A single application of the test material to the non-irrigated eye of three rabbits produced no corneal effects, iridial inflammation was noted in one treated eye one hour after treatment only, with moderate conjunctival irritation noted in all treated eyes one hour after treatment with moderate to minimal conjunctival irritation noted in all treated eyes at the 24-hour observation. This was minimal at 48 -hours and which fully reversed at 72-hours. All treated eyes appeared normal at the 72-hour observation. Under the conditions of this study, the test substance is not considered to be irritating to the eye.

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study performed under GLP. All relevant validity criteria were met.
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 (incl. QA statement)
Remarks:
inspected: March 2014; signature: March 2014
Species:
other: bovine
Strain:
not specified
Details on test animals or tissues and environmental conditions:
SOURCE OF COLLECTED EYES
- Source: Bovine eyes; Recognised supplier.
- Number of animals: Not reported.
- Characteristics of donor animals (e.g. age, sex, weight): Not reported.
- Storage, temperature and transport conditions of ocular tissue (e.g. transport time, transport media and temperature, and other conditions): Refrigerated.
- Time interval prior to initiating testing: Not reported.
- indication of any existing defects or lesions in ocular tissue samples: None reported. Eyes were examined on receipt. Only corneas free of damage were used.
- Indication of any antibiotics used: Penicillin and Streptomycin.
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.75 mL
- Concentration (if solution): undiluted
Duration of treatment / exposure:
10 minutes at 32 ± 1ºC
Duration of post- treatment incubation (in vitro):
After treatment the corneas were incubated for 120 ± 10 minutes at 32 ± 1°C. After the completion of the incubation period opacity determination was performed.
Number of animals or in vitro replicates:
Three (3) per test item, or negative or positive controls, respectively.
Details on study design:
SELECTION AND PREPARATION OF CORNEAS: The eyes were macroscopically checked for unacceptable defects prior to preparation. The eyes were also checked following preparation for defects. Those exhibiting defects were discarded. Following mounting: the anterior and posterior chambers of each BCOP holder were filled with complete Eagle’s Minimum Essential Medium (cMEM) and the holders were incubated at 32 ± 1 ºC for a minimum of 1 hour. After the incubation period, the medium was removed from both compartments and replaced with fresh cMEM.

QUALITY CHECK OF THE ISOLATED CORNEAS: The corneas were examined for defects macroscopically.

NUMBER OF REPLICATES: 3 (Triplicate)

NEGATIVE CONTROL USED: 0.9% w/v sodium chloride solution

SOLVENT CONTROL USED (if applicable): Not applicable.

POSITIVE CONTROL USED: Ethanol

APPLICATION DOSE AND EXPOSURE TIME: 0.75 mL and 10 minutes

TREATMENT METHOD: Closed chamber

POST-INCUBATION PERIOD: Yes. Following exposure the holders were incubated, for 120 ± 10 minutes at 32 ± 1°C. A post-treatment opacity reading was taken and each cornea was visually observed.

REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period: At the end of the exposure period the test item and control items were removed from the anterior chamber and the cornea was rinsed three times with fresh MEM containing phenol red before a final rinse with complete MEM without phenol red. The anterior chamber was refilled with fresh complete MEM without phenol red. A post treatment opacity reading was taken and each cornea was visually observed.
- POST-EXPOSURE INCUBATION: Following the final opacity measurement the posterior compartment was refilled with fresh cMEM. The anterior compartment was filled with 1 ml of 4 mg Na-fluorescein/ml cMEM solution Corneas were incubated in a horizontal position for 90 ± 5 minutes at 32 ± 1°C.

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: Measured through light transmission through the cornea quantitatively using an opacitometer
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of microtiter plate reader (OD492)
- Others (e.g, pertinent visual observations, histopathology): Any other pertinent visual observations would be recorded.

SCORING SYSTEM: In Vitro Irritancy Score (IVIS)

DECISION CRITERIA: The mean opacity and mean permeability values (OD492) were used for each treatment group to calculate an in vitro score:
In vitro irritancy score (IVIS) = mean opacity value + (15 x mean OD492 value). A test item that induces an In Vitro Irritancy Score >/=55.1 is defined as an ocular corrosive or severe irritant. A test item with an IVIS
Irritation parameter:
in vitro irritation score
Value:
0.2
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. The corneas treated with the test item were clear post treatment and post incubation. The corneas treated with the negative control item were clear post treatment and post incubation. The corneas treated with the positive control item were cloudy post treatment and post incubation.
DEMONSTRATION OF TECHNICAL PROFICIENCY:

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Yes.
- Acceptance criteria met for positive control: Yes.
- Range of historical values if different from the ones specified in the test guideline:
Ethanol was used for positive control purposes. The test was acceptable if the positive control produced an In Vitro Irritancy Score which fell within two standard deviations of the current historical control data (HCD) mean: 22.4 to 60.3. ACTUAL: PC IVIS Mean = 42.5.

Table 1. Individual and Mean Corneal Opacity and Permeability Measurements

Treatment Cornea number Opacity Permeability    In vitro irritancy score
Pre-treatment Post-treatment Post-incubation Post incubation - Pre-treatment Corrected value Corrected value
Negative control 1 1 3 4 3 0.044 1.5
2 1 1 1 0  - 0.028
3 1 1 1 0  - 0.036
Mean  - 1.0* 0.036#1
Positive control 1 2 36 34 31.0 31.0 0.846 0.810 42.5
2 1 26 28 26.0 26.0 1.629 1.593
3 1 24 25 24.0 23.0 0.808 0.772
Mean  - 26.7 #2 1.058 #2
Test substance 1 3 3 3 0 0.0 0.053 0.017 0.2
2 1 1 1 0 0.0 0.031 0.000
3 1 1 2 1 0.0 0.066 0.030
Mean 0.0 #2 0.016 #2

OD = Optical Density

* = mean of the post-incubation

#1 = mean permeability

#2 = mean corrected value

Interpretation of results:
GHS criteria not met
Remarks:
EU criteria not met
Conclusions:
Under the conditions of this in vitro study, the test item is not considered to be irritating to the eye.
Executive summary:

The study was performed to OECD TG 437 under GLP to assess the eye irritancy potential of the test item in isolated bovine corneas. The ocular irritancy of the test item was tested through topical application for 10 ± 1 minutes. Negative and positive control items were tested concurrently. The negative control responses for opacity and permeability were less than the upper limits of the laboratory historical range indicating that the negative control did not induce irritancy on the corneas. The mean in vitro irritancy score of the positive control (ethanol), was 42.5 and was within the historical positive control data range (22.4 to 60.3). The test substance did not induce ocular irritation through both endpoints, resulting in a mean in vitro irritancy score of 0.2 after 10 minutes of treatment. Under the conditions of this study, the substance is not considered to be an irritant in the Bovine Corneal Opacity and Permeability test.

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

Respiratory irritation

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

Additional information

Skin Irritation:

In vivo, OECD TG 404, 2014: The study was performed to OECD TG 404 and EU Method B.4 in accordance with GLP to assess the primary skin irritancy potential of the test item in New Zealand White rabbits to assess the irritancy potential of the test item following single, 3-Minute, 1 and 4-Hour, semi-occluded applications to the intact rabbit skin. 0.5 ml of the test material was introduced under a 2.5 cm x 2.5 cm cotton gauze patch and placed in position on the clipped skin. The patch was secured in position with a strip of surgical adhesive tape. 3-Minute and 1-Hour semi-occluded applications of the test item to the intact skin of one rabbit produced no evidence of skin irritation. After 4 hours of exposure to the test substance, the patches were removed and individual dose sites were scored according to the Draize scoring system at approximately 1, 24, 48, and 72 hours. A single 4-Hour, semi occluded application of the test item to the intact skin of three rabbits produced no evidence of skin irritation. Mean scores for following grading at 24, 48 and 72h were zero in both scoring criteria and there was no other signs or systemic toxicity noted during the study. Under the conditions of the study, the substance cannot be considered a skin irritant.

 

Disregarded study: in vitro, OECD TG 439, 2014: The study was performed to OECD TG 439 and EU Method B.46 to assess the skin irritation potential of the test material to a human three dimensional epidermal model. The study was performed under GLP (however not GLP claim is made). Preliminary testing indicated that the test item was MTT-interacting. Therefore appropriate water-killed treated and untreated controls were utilised to determine non-specific MTT reduction. The relative mean tissue viability obtained after 15 minutes treatment with the test substance compared to the negative control tissues was 37.1%. Since the non-specific MTT reduction exceeded 30% the test item is considered incompatible with the test system and no prediction on skin irritation can be made.

 

Eye Irritation:

In vivo, OECD TG 405, 2014: The study was performed to OECD TG 405 and EU Method B.5 under GLP to assess the irritancy potential of the test material to the eye following a single application in the New Zealand White rabbit. A volume of 0.1 ml of the test material was placed into the conjunctival sac of one eye of 3 animals. The other eye remained untreated and was used for control purposes. Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment. A single application of the test material to the non-irrigated eye of three rabbits produced no corneal effects, iridial inflammation was noted in one treated eye one hour after treatment only, with moderate conjunctival irritation noted in all treated eyes one hour after treatment with moderate to minimal conjunctival irritation noted in all treated eyes at the 24-hour observation. This was minimal at 48 -hours and which fully reversed at 72-hours. All treated eyes appeared normal at the 72-hour observation. Under the conditions of this study, the test substance is not considered to be irritating to the eye.

 

In vitro, OECD TG 437, 2014: The study was performed to OECD TG 437 under GLP to assess the eye irritancy potential of the test item in isolated bovine corneas. The ocular irritancy of the test item was tested through topical application for 10 ± 1 minutes. Negative and positive control items were tested concurrently. The negative control responses for opacity and permeability were less than the upper limits of the laboratory historical range indicating that the negative control did not induce irritancy on the corneas. The mean in vitro irritancy score of the positive control (ethanol), was 42.5 and was within the historical positive control data range (22.4 to 60.3). The test substance did not induce ocular irritation through both endpoints, resulting in a mean in vitro irritancy score of 0.2 after 10 minutes of treatment. Under the conditions of this study, the substance is not considered to be an irritant in the Bovine Corneal Opacity and Permeability test.

 

Respiratory Irritation:

In vivo, OECD TG 436, 2013: The study was performed according to OECD TG 436 and in accordance with GLP to assess the acute inhalation toxicity of the test substance. A group of Wistar rats comprising three males and three females was exposed to an aerosol atmosphere for four hours using a nose only exposure system, followed by a fourteen day observation period. The target concentration was 5.0 mg/L and the mean achieved atmosphere concentration was 5.11 mg/L. There was no reports of nasal irritation and the respiratory tract was subjected to a detailed macroscopic examination for signs of irritancy or local toxicity during necropsy; none was reported post exposure.

Justification for classification or non-classification

The substance does not meet classification criteria under Regulation (EC) No 1272/2008 for dermal irritation. 

The substance does not meet classification criteria under Regulation (EC) No 1272/2008 for eye irritation.

The substance does not meet classification criteria under Regulation (EC) No 1272/2008 for Specific Target Organ Toxicant: STOT-SE category 3: respiratory irritation or narcotic effects.

For skin irritation, the weight of evidence indicates that the substance has the potential to be slightly irritating however this is transient and not sufficient for classification purposes. Available data indicates negative irritation in an in vivo dermal irritation study. This study utilised semi-occlusive dressing. Applicant recalculation of results: The individual scoring results and observations were presented in the full study report the results according to the EU criteria of CLP Regulation (EC) 1272/2008 as amended indicates that the mean Erythema and Oedema scores are not ≥ 2.3 - ≤ 4.0 within the 24 to 72 hour period in 2 of 3 organisms. Any effects fully reversed within the observation period of the study and there was an absence of pronounced variability of responses. There was an absence of significant other effects such as alopecia (limited) area or hyperkeratosis or hyperplasia and scaling in all related studies (acute dermal toxicity and skin sensitisation). Particularly any such effects remaining at the end of the observation period. The substance would not be considered a skin irritant. The test substance was considered incompatible with and available RHE in vitro assay under guideline and this result is not further considered.

For eye irritation, the weight of evidence indicates that the substance has the potential to cause transient mild irritating effects to the eye but which are insufficient for classification based on the applicants recalculation of the mean scoring and evaluation of the results in three organisms demonstrating that the EU criteria had not been met. Effects in vivo on corneal opacity are non-existent and iritis and conjunctival effects are very low which fully reversed within 72 hours; the overall evidence is indicative of transient and reversible effects on the eye and which is supported with an in vitro bovine corneal assay indicating no irritation.

For respiratory irritation (Specific Target Organ Toxicant: STOT-SE category 3: respiratory irritation or narcotic effects), the weight of evidence indicates from an available OECD TG 436 study with additional examinations of the respiratory tract at the EU Criteria limit dose (5.0 mg/L) that there was no irritation to the nasal area or respiratory related clinical signs reported or narcotic effects (ataxia or related clinical signs) at relevant maximum dose-concentration guidance values for mists. There is no evidence that the substance meets the classification criteria in relevant animals studies. The assessment is made on animal data only in accordance with Regulation (EC) No. 1272/2008 as amended Annex 1: 3.8.2.1.10.1. No relevant human data is available.

References:

1. ECHA Guidance on Application on the CLP Criteria, section 3.2 (version 5.0, July 2017)

2. ECHA Guidance on Information Requirements and Chemical Safety Assessment (Chapter R.7a: Endpoint Specific Guidance, R.7.2.2, version 6.0, July 2017)