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Description of key information

Skin irritation

Key, rabbit, OECD 404, GLP: negative

Eye irritation

Key, rabbit, OECD 405, GLP: negative

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:
8 JUL 1996 - 30 SEP 1996
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
This study was conducted in compliance with the regulations for Good Laboratory Practice (GLP) in nonclinical laboratory studies and followed the OECD Guideline for Testing of Chemicals No 404.
Qualifier:
according to guideline
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Strain: Albino rabbit, Crl:KBL(NZW)BR, female (f)
- Source: Charles River Wiga GmbH, Kißlegg
- Age at study initiation: about 16 weeks
- Weight at study initiation: 2.80 (2.51 - 2.96) kg
- Housing: They were kept separately in cages type KK 017 (floor area: 75 x 54.5 = 4087 cm2, height: 40 cm; manufacturer: Hulskamp) on metal grids. The paper underneath the cages was changed three times a week. The cages were machine-cleaned before the start of the study.
- Diet (e.g. ad libitum): ad libitum, Altromin Standard Diet TPF(R) 2113, pellets and acidified (The diet is checked periodically according to the specifications of the manufacturer by an independent laboratory, approved by the German government. Analysis includes both qualitative and quantitative evaluation for heavy metals, aflatoxins, pesticides, and antibiotics. The analytical results show that the limits set by German regulations for animal feed have been followed.)
- Water (e.g. ad libitum): ad libitum, fully demineralized water
- Acclimation period: more than 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 21°C
- Humidity (%): 48 to 93
- Air changes (per hr): air-conditioned
- Photoperiod (hrs dark / hrs light): daylight
Type of coverage:
semiocclusive
Preparation of test site:
shaved
Vehicle:
water
Controls:
not required
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 g
- Concentration (if solution): 0.5 g test item was mixed with some drops aqua pro injectione

Duration of treatment / exposure:
4 hours
Observation period:
The rabbits were investigated for skin alterations, behaviour, and general condition 1 hour after removal of the patches, and then after 24, 48, 72 hours and then daily up to day 8 of the study.
Number of animals:
3 female rabbits
Details on study design:
TEST SITE
- Area of exposure: 6 cm2
- Type of wrap if used: 6 cm2 patches. The patch was kept in place by a self-adhesive fabric, Fixomull® stretch, Beiersdorf.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Any test material left was wiped off after removal of the patches.
- Time after start of exposure: 4 h

OBSERVATION TIME POINTS
1 h, 24 h, 48 h, 72 h after removal of the patches and the daily until day 8 of the study

SCORING SYSTEM:
- Method of calculation:

Grading scale for evaluation:

Erythema and eschar formation

Scores
--------------------------------------------------
No erythema 0

Very slight erythema (barely perceptible) 1

Well defined erythema 2

Moderate to severe erythema 3

Severe erythema (beet redness) to eschar
formation preventing grading of erythema 4
--------------------------------------------------
Maximum possible: 4
--------------------------------------------------

Edema formation
Scores
--------------------------------------------------
No edema 0

Very slight edema (barely perceptible) 1

Slight edema (edges of area well
defined by definite raising) 2

Moderate edema (raised approx. 1 mm) 3

Severe edema (raised more than 1 mm
and extending beyond area of exposure) 4
--------------------------------------------------
Maximum possible: 4
--------------------------------------------------

Total possible irritation score (maximum): 8

Mean score = Mean grading for erythema or edema of all the rabbits per time point
Mean cumulative score = Mean grading of all readings for erythema and edema per time point
Mean score per animal = Mean grading for each animal of erythema or edema (24, 48, and 72 hours after removal of the patches)
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
other: 1 h, 24 h, 48 h, 72 h
Score:
0
Max. score:
8
Reversibility:
other: no effects
Remarks on result:
no indication of irritation
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: no effects
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
Reversibility:
other: no effects
Remarks on result:
no indication of irritation
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: no effects
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
Reversibility:
other: no effects
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
Reversibility:
other: no effects
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
Reversibility:
other: no effects
Remarks on result:
no indication of irritation
Irritant / corrosive response data:
After a single application to the intact dorsal skin of rabbits, for a 4 hours period of exposure under semiocclusive conditions, no signs of irritation were observed in the treated areas.
Other effects:
None

Clinical findings

No signs of systemic toxicity were detected.

Mortality

All rabbits survived the 8 day observation period.

Body weight

Body weight development of the treated rabbits was inconspicuous.

Interpretation of results:
GHS criteria not met
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
According to the result, the test material is not irritant to the skin.
Executive summary:

The registered substance was tested for skin irritation in an Acute Dermal Irritation / Corrosion test according to OECD Guideline 404 following GLP.

Purpose

The purpose of this assay was to identify the skin irritation/corrosion potential of the test item when applied to the intact skin of previously shaven rabbits for a 4 hours period under semiocclusive conditions. This study should provide a rational basis for risk assessment to the irritating potential of the test item in human.

Study design

To test for primary skin irritation, the test substance was mixed with some drops of aqua pro injectione to ensure good contact to the skin. Afterwards the test material was spread onto patches and applied to the intact skin of three previously shaven rabbits for a 4 hours period under semiocclusive conditions. The first examination of the treated skin sites followed 1 hour after removal of the patches. Then, examinations were performed daily for further 7 days.

Results

Under the conditions of the present study no signs of irritation were seen. The evaluation of all animals yields a zero mean score.

Conclusion

According to the result, the test material is not irritant to the skin.

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:
8 JUL 1996 - 25 SEP 1996
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
This study was conducted according to Good Laboratory Practice (GLP) and followed the OECD guideline for testing of Chemicals 405.
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Strain: Albino rabbit, Crl:KBL(NZW)BR, female (f)
- Source: Charles River Wiga GmbH, Kißlegg
- Age at study initiation: about 16 weeks
- Weight at study initiation: 2.84 (2.78 - 2.88) kg
- Housing: They were kept separately in cages type KK 017 (floor area: 75 x 54.5 = 4087 cm2, height: 40 cm; manufacturer: Hulskamp) on metal grids. The paper underneath the cages was changed three times a week. The cages were machine-cleaned before the start of the study.
- Diet (e.g. ad libitum): ad libitum, Altromin Standard Diet TPF(R) 2113, pellets and acidified (The diet is checked periodically according to the specifications of the manufacturer by an independent laboratory, approved by the German government. Analysis includes both qualitative and quantitative evaluation for heavy metals, aflatoxins, pesticides, and antibiotics. The analytical results show that the limits set by German regulations for animal feed have been followed.)
- Water (e.g. ad libitum): ad libitum, fully demineralized water
- Acclimation period: more than 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 21°C
- Humidity (%): 48 to 93
- Air changes (per hr): air-conditioned
- Photoperiod (hrs dark / hrs light): daylight
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 g
Duration of treatment / exposure:
0.1 g test material was instilled into the conjunctival sac of the left eye of each animal by gently pulling the lower lid away from the eyeball to form a cup into which the test material was placed. The right eye remained untreated and served as control. After instillation, the eyelids were closed for about 30 seconds.
Observation period (in vivo):
The rabbits were investigated for eye irritation and for changes in behaviour and general condition 1 hour after treatment, and than after 24, 48, 72 hours and so on daily up to day 8 of the experimental part.
Number of animals or in vitro replicates:
3 female rabbits
Details on study design:
SCORING SYSTEM:

-- Evaluation of eye reactions

- Cornea Scores
A) Opacity-degree of density (area most dense taken for reading)

No ulceration or opacity 0

Scattered or diffuse areas of opacity (other than slight dulling
of normal lustre), details of iris clearly visible 1
Easily discernible translucent area, details of iris slightly obscured 2
Nacrous area, no details of iris visible, size of pupil barely
discernible 3
Opaque cornea, iris not discernible through the opacity 4

B) Area of cornea involved not evaluated (Rating according to the DRAIZE method)

- Iris
A) Normal 0
Markedly deepened rugae, congestion, swelling, moderate
circumcorneal hyperemia, or injection, any of these or combination
of any thereof, iris still reacting to light
(sluggish reaction is positive) 1
No reaction to light, hemorrhage, gross destruction
(any or all of these) 2

- Conjunctivae Scores
A) Redness (refers to palpebral and bulbar conjunctivae,
cornea and iris)
Blood vessels normal 0
Some blood vessels definitely hyperemic (injected) 1
Diffuse, crimson color, individual vessels not easily discernible 2
Diffuse beefy red 3

B) Chemosis: lids and/or nictating membranes
No swelling 0
Any swelling above normal (includes nictating membranes) 1
Obvious swelling with partial eversion of lids 2
Swelling with lids about half closed 3
Swelling with lids more than half closed 4

C) Discharge (Rating according to the DRAIZE method )
No discharge 0
Any amount different from normal (does not include small
amounts observed in inner canthus of normal animals) 1
Discharge with moistening of the lids and hairs just adjacent to lids 2
Discharge with moistening of the lids and hairs,
and of a considerable area around the eye 3

Mean score of all animals = Mean grading for irritations of cornea, iris, and conjunctivae per time point
Mean score per animal = Mean grading for each animal of irritations of cornea, iris, and conjunctivae (1, 24, 48, and 72 hours after application)
Maximum value = Maximum grading of a sign of irritation within a period
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: no effects
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
Reversibility:
other: no effects
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
Reversibility:
other: no effects
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:
other: no effects
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
Reversibility:
other: no effects
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
Reversibility:
other: no effects
Remarks on result:
no indication of irritation
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0.33
Max. score:
3
Reversibility:
fully reversible within: 48 h
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
3
Reversibility:
other: no effects
Remarks on result:
no indication of irritation
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
3
Reversibility:
other: no effects
Remarks on result:
no indication of irritation
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: no effects
Remarks on result:
no indication of irritation
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: no effects
Remarks on result:
no indication of irritation
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: no effects
Remarks on result:
no indication of irritation
Irritant / corrosive response data:
After single instillation of 0.1 g test material, into the conjunctival sac of the left eye, no signs of irritation of the cornea and iris were observed. The conjunctivae showed redness (score 1) at the first reading and in one animal also at the second reading. Chemosis (scores 1 and 2) was seen in two animals at the first reading. Later on, no signs of irritation were seen.
The untreated eyes were normal.
Other effects:
No signs of clinical toxicity were detected.

Ocular alterations

After single instillation of 0.1 g test material, into the conjunctival sac of the left eye, no signs of irritation of the cornea and iris were observed. The conjunctivae showed redness (score 1) at the first reading and in one animal also at the second reading. Chemosis (scores 1 and 2) was seen in two animals at the first reading. Later on, no signs of irritation were seen.

Clinical findings

No signs of irritation were observed at the cornea and iris. The conjunctivae showed redness (score 1) on day 1 of the study. Later on no signs of irritation were observed. The untreated eyes were unchanged.

No signs of clinical toxicity were detected.

Mortality

All animals survived the observation period.

Body weight

Body weight development of the treated rabbits was inconspicuous.

Interpretation of results:
GHS criteria not met
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
According to the result of the irritation scores, the test material has not to be classified as an eye irritant.
Executive summary:

The registered substance was tested for eye irritation in an Acute Eye Irritation / Corrosion test according to OECD Guideline 405 following GLP.

Purpose
The purpose of this primary eye irritation assay was to provide information on possible health hazards in case of acute eye contact with a test material and serve as a a rational basis for risk assessment to the eye irritating potential of the test item in human.

Study design
The test material was tested for eye irritation potential. 0.1 g of the test material was applied into the conjunctival sac of rabbits. The first examination of the eyes followed 1 hour after instillation. The examinations were performed daily for further 7 days.

Results
No signs of irritation concerning to cornea and iris were observed. The conjunctivae showed redness (score 1), and chemosis (scores 1 and 2) at the first and second readings, maximally. This is a common finding in the case of powder. The untreated eyes were unchanged.

The cumulative mean scores/group for cornea, iris, and conjunctivae were:

Study day
(hours)
1
(1)
2
(24)
3
(48)
4
(72)
Cornea                                 0.0 0.0 0.0 0.0
Iris                                         0.0 0.0 0.0 0.0
Conjunctivae                     redness
                                             chemosis
1.0
1.0
0.3
0.0
0.0
0.0
0.0
0.0


The scores/animal (24, 48, 72 hours) were:

Mean Max
Animal No. 14 13 12 14 13 12
Cornea                                 0.0 0.0 0.0 0 0 0
Iris                                         0.0 0.0 0.0 0 0 0
Conjunctivae                     redness
                                             chemosis
0.3
0.0
0.0
0.0
0.0
0.0
1
0
0
0
0
0



Conclusion
According to the result of the irritation scores, the test material has not to be classified as an eye irritant.

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

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation

Acute Dermal Irritation / Corrosion test, OECD 404

The registered substance was tested for skin irritation in an Acute Dermal Irritation / Corrosion test according to OECD Guideline 404 following GLP.

Purpose

The purpose of this assay was to identify the skin irritation/corrosion potential of the test item when applied to the intact skin of previously shaven rabbits for a 4 hours period under semiocclusive conditions. This study should provide a rational basis for risk assessment to the irritating potential of the test item in human.

Study design

To test for primary skin irritation, the test substance was mixed with some drops of aqua pro injectione to ensure good contact to the skin. Afterwards the test material was spread onto patches and applied to the intact skin of three previously shaven rabbits for a 4 hours period under semiocclusive conditions. The first examination of the treated skin sites followed 1 hour after removal of the patches. Then, examinations were performed daily for further 7 days.

Results

Under the conditions of the present study no signs of irritation were seen. The evaluation of all animals yields a zero mean score.

Conclusion

According to the result, the test material is not irritant for the skin.

Eye irritation

Acute Eye Irritation / Corrosion, OECD 405

The registered substance was tested for eye irritation in an Acute Eye Irritation / Corrosion test according to OECD Guideline 405 following GLP.

Purpose
The purpose of this primary eye irritation assay was to provide information on possible health hazards in case of acute eye contact with a test material and serve as a a rational basis for risk assessment to the eye irritating potential of the test item in human.

Study design
The test material was tested for eye irritation potential. 0.1 g of the test material was applied into the conjunctival sac of rabbits. The first examination of the eyes followed 1 hour after instillation. The examinations were performed daily for further 7 days.

Results
No signs of irritation concerning to cornea and iris were observed. The conjunctivae showed redness (score 1), and chemosis (scores 1 and 2) at the first and second readings, maximally. This is a common finding in the case of powder. The untreated eyes were unchanged.

The cumulative mean scores/group for cornea, iris, and conjunctivae were:

Study day
(hours)
1
(1)
2
(24)
3
(48)
4
(72)
Cornea                                 0.0 0.0 0.0 0.0
Iris                                         0.0 0.0 0.0 0.0
Conjunctivae                     redness
                                             chemosis
1.0
1.0
0.3
0.0
0.0
0.0
0.0
0.0


The scores/animal (24, 48, 72 hours) were:

Mean Max
Animal No. 14 13 12 14 13 12
Cornea                                 0.0 0.0 0.0 0 0 0
Iris                                         0.0 0.0 0.0 0 0 0
Conjunctivae                     redness
                                             chemosis
0.3
0.0
0.0
0.0
0.0
0.0
1
0
0
0
0
0



Conclusion

According to the result of the irritation scores, the test material has not to be classified as an eye irritant.


Justification for classification or non-classification

Based on the results of the key studies for in vivo skin and eye irritation, no classification for irritation is triggered in accordance with Regulation (EC) No 1272/2008.