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Administrative data

Description of key information

Skin irritation, in vivo (rabbit): non-irritating, OECD TG 404, 2000

Supporting study: Skin Irritation, in vivo (rat): mild transient irritation, OECD TG 402, 2008

Eye irritation, in vivo (rabbit): non-eye irritating, OECD TG 405, 2000

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:
11-04-2000 to 21-07-2000
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:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
inspected: September 1999 ; signature: December 1999
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Recognised animal supplier
- Age at study initiation: Young adult
- Weight at study initiation: 2.4 ±0.1 kg
- Housing: Individually housed in polystyrene cages (48.2 cm x 58 cm x 36.5 cm).
- Diet: certified rabbit diet ad libitum
- Water: filtered certified water ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 ±3
- Humidity (%): 30 - 70
- Air changes (per hr): ca. 12 cycles/hr
- Photoperiod: 12 hours light / 12 hours dark

IN-LIFE DATES: From: 11-04-2000 To: 25-04-2000
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): Not applicable.
Duration of treatment / exposure:
4 hours
Observation period:
72 hours (initial observation); additional observations are made daily up to and including on Days 7 and 15 to assess the reversibility of skin reactions (as appropriate).
Number of animals:
3
Details on study design:
TEST SITE
- Area of exposure: dorsal
- Type of wrap if used: semi-occlusive ; gauze patch secured with semi-occlusive dressing and restraining bandage

REMOVAL OF TEST SUBSTANCE
- Washing (if done): No, since no residual test item was observed following dressing removal.
- Time after start of exposure: 4 hours

OBSERVATION TIME POINTS
1, 24, 48, 72 hours (initial observations); additional observations are made daily up to and including on Days 7 and 15 to assess the reversibility of skin reactions (as appropriate).

SCORING SYSTEM:
Consistent with Draize. 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 (clinical signs/behaviour), if present, were also recorded.
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
other: 1h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 9 days
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
other: 1h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 8 days
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
other: 1h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 8 days
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 9 days
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
24/48/72 h
Score:
1.3
Max. score:
4
Reversibility:
fully reversible within: 8 days
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 8 days
Irritation parameter:
edema score
Basis:
animal #1
Time point:
other: 1h
Score:
0
Max. score:
4
Remarks on result:
no indication of irritation
Irritation parameter:
edema score
Basis:
animal #2
Time point:
other: 1h
Score:
0
Max. score:
4
Remarks on result:
no indication of irritation
Irritation parameter:
edema score
Basis:
animal #3
Time point:
other: 1h
Score:
0
Max. score:
4
Remarks on result:
no indication of irritation
Irritation parameter:
edema score
Basis:
animal #1
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:
edema score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
4
Remarks on result:
no indication of irritation
Irritant / corrosive response data:
- Erythema: Very slight or well defined erythema (score = 1 or 2) at all treated skin sites at 24 to 72 hours after patch removal and up to day 7 or 8 in two sites.
- Edema: No evidence of skin irritation was noted during the study.
- Reversibility of effects: All effects reversed within 9 days (3/3 sites) ; dryness of the skin was noted in all sites from day 5 (1/3), day 6 (2/3) or day 9 (1/3) which ceased in 2/3 sites by day 14.
Other effects:
- Other adverse local effects: None reported
- Other adverse systemic effects: None reported.

Table 1. Individual skin reactions

Skin Reaction

Observation Time

Individual Scores

 

 

1 (male)

2 (male)

3 (male)

Erythema/Eschar formation

1 hour

1

1

1

 

24 hours

2

2

2

 

48 hours

2

1

2

 

72 hours

2

1 S

2

 

7 days

1

1 S

1 S

 

14 days

0

0

0 S

 

 

 

 

 

Oedema formation

1 hour

0

0

0

 

24 hours

0

0

0

 

48 hours

0

0

0

 

72 hours

0

0

0

 

7 days

0 S

0 S

0

 

14 days

0

0

0 S

 

 

 

 

 

S = dryness of the skin

 

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

Erythemea/Escar Formation:

1: total = 6; mean score = 2.0

2: total = 4; mean score = 1.3

3: total = 6; mean score = 2.0

Oedema Formation:

1: total = 0; mean score = 0.0

2. total = 0; mean score = 0.0

3. total = 0; mean score = 0.0

Interpretation of results:
GHS criteria not met
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
Under the conditions of this study the test item is not considered to be irritating.
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. Following single 4-Hour, semi-occluded applications to the intact rabbit skin. 0.5 ml of the test material was introduced under a gauze patch and placed in position on the clipped skin via semi-occlusive dressing to assess the irritancy potential of the test item. The patch was secured in position with a strip of surgical bandage. After 4 hours of exposure to the test item, the patches were removed, and individual dose sites were scored at approximately 1, 24, 48, and 72 hours. Daily observations were made up to day 15 as applicable. A single 4-Hour, semi occluded application of the test item to the intact skin of three rabbits produced very slight or well-defined erythema (score = 1 or 2) at all treated skin sites at 24 to 72 hours after patch removal and up to day 7 or 8 in two sites. Mean scores for following grading at 24, 48 and 72h were 2.0, 1.3 and 2.00 in erythema and eschar and 0.0 in all sites for the edema scoring criteria. Under the conditions of the study, the test item is not considered to be a skin irritant.

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:
18-04-2000 to 19-07-2000
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:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
inspected: September 1999 ; signature: December 1999
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: Young adult
- Weight at study initiation: 2.6 ±0.1 kg
- Housing: Individually housed in polystyrene cages (48.2 cm x 58 cm x 36.5 cm).
- Diet: certified rabbit diet ad libitum
- Water: filtered certified water ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 ±3
- Humidity (%): 30 - 70 (the relative humidity was stated as being periodically outside this range ; this was not considered to impact the study integrity).
- Air changes (per hr): ca. 12 cycles/hr
- Photoperiod: 12 hours light / 12 hours dark

IN-LIFE DATES: From: 18-04-2000 To: 22-04-2000
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 left 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. The right eye remained untreated and was used for control purposes.
Observation period (in vivo):
Ocular assessment was conducted at approximately 1, 24, 48 and 72 hours after instillation of the test item, according to numerical evaluation.
Number of animals or in vitro replicates:
3 (male).
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): None
- Time after start of exposure: Not applicable.

SCORING SYSTEM:
The irritation was assessed using criteria consistent with Draize (1977) numerical scoring system. At each observation period, the highest scores given were recorded. Any other ocular effects were also noted.

TOOL USED TO ASSESS SCORE: Examination of the eye was facilitated by the use of the light source from a standard ophthalmoscope (or if corneal opacification is difficult to determine using a UV lamp).
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
24/48/72 h
Score:
0
Max. score:
4
Remarks on result:
other: mean; n=2
Irritation parameter:
iris score
Basis:
mean
Time point:
24/48/72 h
Score:
0.1
Max. score:
2
Reversibility:
fully reversible within: 72 hours
Remarks on result:
other: mean; n=3
Irritation parameter:
conjunctivae score
Basis:
mean
Time point:
24/48/72 h
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 72 hours
Remarks on result:
other: mean; n=3
Irritation parameter:
chemosis score
Basis:
mean
Time point:
24/48/72 h
Score:
0.3
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Remarks on result:
other: mean; n=3
Irritant / corrosive response data:
No corneal or Iridial inflammation effects were noted. Very slight conjunctival reactions (chemosis and redness, score = 1) was noted in one treated eyes 1 and 24 hours after treatment. Although chemosis fully resolved by 48 hour observation. A very slight redness (score = 1) persisted until day 4 in 1/3 treated eyes.
Other effects:
- Lesions and clinical observations: None reported.
- Ophthalmoscopic findings: None reported using fluorescein.
- Histopathological findings: None reported.
- Effects of rinsing or washing: Not applicable.
- Other observations: None reported.

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

1

0

1

 

24 h

0

0

0

0

 

48 h

0

0

0

0

 

72 h

0

0

0

0

 

Day 4

-

-

-

-

Mean (24 – 72 h)

 

0.00

0.00

0.00

0.00

 

 

 

 

 

 

2 (male)

1h

0

0

0

1

 

24 h

0

0

0

0

 

48 h

0

0

0

0

 

72 h

0

0

0

0

 

Day 4

-

-

-

-

Mean (24 – 72 h)

 

0.00

0.00

0.00

0.00

 

 

 

 

 

 

3 (male)

1h

0

0

1

1

 

24 h

0

0

1

1

 

48 h

0

0

1

0

 

72 h

0

0

1

0

 

Day 4

0

0

0

0

Mean (24 – 72 h)

 

0.00

0.00

1.0

0.33

 

 

 

 

 

 

Interpretation of results:
GHS criteria not met
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
Under the conditions of this study the test item 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 item to the eye following a single application in the New Zealand White rabbit. A volume of 0.1 ml of the test item was placed into the conjunctival sac of one eye of three 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 item to the non-irrigated eye of three rabbits produced no corneal or Iridial inflammation effects. Very slight conjunctival reactions (chemosis and redness, score = 1) was noted in one treated eyes 1 and 24 hours after treatment. Although chemosis fully resolved by 48 hour observation. A very slight redness (score = 1) persisted until day 4 in 1/3 treated eyes. No other ocular reactions were reported during the study. All treated eyes appeared normal at the 4 day observation point. Under the conditions of this study, the test item is not considered to be irritating to the eye.

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

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin Irritation:

Key study: In vivo, OECD TG 404, 2000: 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. Following single 4-Hour, semi-occluded applications to the intact rabbit skin. 0.5 ml of the test material was introduced under a gauze patch and placed in position on the clipped skin via semi-occlusive dressing to assess the irritancy potential of the test item. The patch was secured in position with a strip of surgical bandage. After 4 hours of exposure to the test item, the patches were removed, and individual dose sites were scored at approximately 1, 24, 48, and 72 hours. Daily observations were made up to day 15 as applicable. A single 4-Hour, semi occluded application of the test item to the intact skin of three rabbits produced very slight or well-defined erythema (score = 1 or 2) at all treated skin sites at 24 to 72 hours after patch removal and up to day 7 or 8 in two sites. Mean scores for following grading at 24, 48 and 72h were 2.0, 1.3 and 2.00 in erythema and eschar and 0.0 in all sites for the edema scoring criteria. Under the conditions of the study, the test item is not considered to be a skin irritant.

Supporting study: In vivo, OECD TG 402, 2008: The study was performed according to OECD TG 402 and EU Method B.3 Acute Toxicity (Dermal) and in accordance with GLP to assess the acute dermal toxicity of the test substance in the Wistar HanRcc: WIST (SPF) strain rat. A group of ten animals (five males and five females) was given a single, 24 hour, semi-occluded dermal application of the diluted test item in PEG 300 vehicle to intact skin at a dose level of 2000 mg/kg body weight. Clinical signs and body weight development were monitored during the study. All animals were subjected to gross necropsy. There was no mortality during the study. There were no signs of system toxicity or abnormalities on necropsy. Animals showed expected gains in body weight. Minimal signs of dermal irritation were noted (score 2) at day 2 and (score 1) days 4 to 5 or 6, which had fully reversed at day 7. The dermal LD50 was established to exceed 2000 mg/kg bw in male/female Wistar HanRcc: WIST (SPF) rat. Under the conditions of this study, and according to the GHS criteria, the LD50 cut-off value was considered to be greater than 5000 mg/kg body weight.

Eye Irritation:

Key study: In vivo, OECD TG 405, 2000: The study was performed to OECD TG 405 and EU Method B.5 under GLP to assess the irritancy potential of the test item to the eye following a single application in the New Zealand White rabbit. A volume of 0.1 ml of the test item was placed into the conjunctival sac of one eye of three 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 item to the non-irrigated eye of three rabbits produced no corneal or Iridial inflammation effects. Very slight conjunctival reactions (chemosis and redness, score = 1) was noted in one treated eyes 1 and 24 hours after treatment. Although chemosis fully resolved by 48 hour observation. A very slight redness (score = 1) persisted until day 4 in 1/3 treated eyes. Mean scores calculated for each animal over 24, 48 and 72 hours were 0.0, 0.0 and 0.3 for chemosis, 0.0, 0.0 and 1.0 for redness of the conjunctiva, 0.0, 0.0 and 0.0 for iris lesions and 0.0, 0.0 and 0.0 for corneal opacity. No other ocular reactions were reported during the study. All treated eyes appeared normal at the 4 day observation point. Under the conditions of this study, the test item is not considered to be irritating to the eye.

Respiratory Irritation:

No study available

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.

 

For skin irritation, further in vitro skin corrosion testing does not need to be conducted based on the available information allowing a definitive conclusion on the classification of the substance. The substance does not demonstrate significant skin irritation potential necessary for classification and labelling within an available skin irritation in vivo assay (OECD TG 404) and supporting acute dermal toxicity study (OECD TG 402) at 2000 mg/kg bw dose level. The substance can cause pronounced transient mild irritation but which is not sufficient for classification and labelling.

 

For eye irritation, the weight of evidence indicates that the substance has the potential to cause transient irritating effects to the eye but which is not sufficient for classification based on 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 and iritis are low to non-existent and conjunctival effects are low to moderate which fully reversed within 72 hours to 96 hours; the overall evidence is indicative of transient mild effects on the eye.

References:

1. Guidance on Application of the CLP Criteria, ECHA, version 5.0, July 2017