Registration Dossier

Administrative data

Description of key information

Skin irritation (OECD 404), rabbit: irritating

Eye irritation (OECD 405), rabbit: corrosive

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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Version / remarks:
adopted in 2002
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
Version / remarks:
adopted in May 2008
Deviations:
no
GLP compliance:
yes
Species:
rabbit
Strain:
other: SPF albino of the stock Chbb:HM
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland GmbH, Kißlegg, Germany
- Age at study initiation: approx. 20-21 months
- Weight at study initiation: 2.7-2.9 kg
- Housing: animals were caged individually in PPO caged with perforated floor
- Diet: pelleted complete rabbit diet “Altromin 2123”, ad libitum
- Water: tap water, ad libitum
- Acclimation period: at least one week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3
- Humidity (%): 30-70
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12
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): 0.5 g


Duration of treatment / exposure:
4 h (confirmatory test)
Observation period:
14 days
Reading time points: 1, 24, 48, and 72 h and 7 and 14 days
Number of animals:
3 females
Details on study design:
TEST SITE
- Area of exposure: anterior right field of the back of each rabbit
- Type of wrap if used: 16- layer gauze patch (2.5 x 2.5 cm) secured semi-occlusively with adhesive Gothaplast tape (2.5 cm wide) and fixed with Gothaplast tape (5 cm wide) loosely wound around the trunk of the animals.
REMOVAL OF TEST SUBSTANCE
- Washing (if done): the treated skin was cleaned with mild soap and lukewarm water.
- Time after start of exposure: 4 h
SCORING SYSTEM: Draize score system
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
not fully reversible within: 14 days
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
not fully reversible within: 14 days
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
not fully reversible within: 14 days
Irritation parameter:
edema score
Basis:
mean
Remarks:
out of all three animals
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritant / corrosive response data:
One animal used in the initial test, showed a well-defined erythema and a very slight edema on the posterior right test field immediately after the application.
1 h after the termination of exposure a well-defined erythema and a very slight edema were noted on the posterior right test field of the same animal. Two further animals displayed a very slight erythema on the anterior right test field.
24, 48 and 72 h after the termination of exposure a well-defined erythema was observed on the posterior right test field of the first animal. A second animal showed a very slight erythema on the anterior right test field, whereas a well-defined erythema was observed on the anterior right test field of the third animal.
7 days after the termination of the application the first animal showed large scales in a middle-grade on the whole posterior right test field. Slight scurf was observed on the anterior right test field of the second and third animals.
14 days after the termination of application the first animal showed slight scurf on then whole posterior right test field. Isolated scales were observed on the anterior right test field of the second animal, whereas the third animal was free of any signs of skin irritation.
Interpretation of results:
other: Skin irrit. 2, H315. Classification according to Regulation (EC) No. 1272/2008 (CLP/EU GHS).
Endpoint conclusion
Endpoint conclusion:
adverse effect observed (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:
24 Nov - 23 Dec 2008
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
adopted in 2002
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Version / remarks:
adopted in 2008
Deviations:
no
GLP compliance:
yes
Species:
rabbit
Strain:
other: SPF albino of the stock Chbb:HM
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland GmbH, Kißlegg, Germany
- Age at study initiation: approximately 15- 33 months
- Weight at study initiation: 2.7 kg (mean)
- Housing: animals were caged individually in PPO caged with perforated floor
- Diet: pelleted complete rabbit diet “Altromin 2123”, ad libitum
- Water: tap water, ad libitum
- Acclimation period: approx. one week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3
- Humidity (%): 30-70
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

Vehicle:
unchanged (no vehicle)
Controls:
other: the untreated eye served as control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 g

Duration of treatment / exposure:
1h
Observation period (in vivo):
21 days
Reading time points: 1, 24, 48 and 72 h and 7 and 14 days
Number of animals or in vitro replicates:
3 females
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): physiologic saline solution
- Time after start of exposure: 1h

SCORING SYSTEM: Draize scoring system

TOOL USED TO ASSESS SCORE: fluorescein
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
not fully reversible within: 7 days
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
not reversible
Remarks:
within 21 days
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
not reversible
Remarks:
within 21 days
Irritation parameter:
iris score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1
Max. score:
2
Reversibility:
fully reversible
Remarks:
within 7 days
Irritation parameter:
iris score
Basis:
animal #2
Time point:
24/48/72 h
Score:
1
Max. score:
2
Reversibility:
fully reversible
Remarks:
within 7 days
Irritation parameter:
iris score
Basis:
animal #3
Time point:
24/48/72 h
Score:
1
Max. score:
2
Reversibility:
fully reversible within: 7 days
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
24/48/72 h
Score:
2
Max. score:
3
Reversibility:
fully reversible within: 14 days
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
24/48/72 h
Score:
2
Max. score:
3
Reversibility:
fully reversible within: 21 days
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
24/48/72 h
Score:
1.33
Max. score:
3
Reversibility:
fully reversible within: 21 days
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 21 days
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
24/48/72 h
Score:
1.33
Max. score:
4
Reversibility:
fully reversible within: 21 days
Irritant / corrosive response data:
One hour after application of the test material one test animal showed punctate or opacity with clearly identifiable details of iris to about a quarter of the cornea, some hyperemia, conjunctival blood vessels to about a swelling more than normal. In two further animals, punctate or diffuse areas of opacity were observed with clearly identifiable details of iris on more than three-quarters of the corneal surface, some hyperemia, conjunctival blood vessels and swelling more than normal.
24 hours after application of the test material all three animals showed punctate or diffuse areas of opacity with clearly identifiable details of iris on more than three-quarters of the surface of the cornea, an iris with significantly deepen rugae, congestion, swelling, moderate circumcorneal hyperemia and existing light reaction, a conjunctiva with diffuse crimson color and difficult to identify individual vessels and a marked swelling with partial eversion the eyelids. In addition, whitish secretion to the entire eye of one animal, and in the corner of the eye, in two further animals was observed.
After the installation of fluorescein animals all three animals showed punctate or diffuse areas of opacity with clearly identifiable details of iris on more than three-quarters of the corneal surface.
48 h after administration of the test material, in one animal punctate or diffuse areas of opacity with clearly identifiable details of iris on more than three-quarters of the surface of the cornea, an iris with significantly deepen rugae, congestion, swelling, moderate circumcorneal hyperemia and existing light reaction, a conjunctiva with diffuse crimson color and difficult to identify individual vessels, a marked swelling with partial eversion of the lids and whitish secretions around the entire eye were observed. A second animal showed punctate or diffuse areas of opacity with clearly identifiable details of iris on more than three-quarters of the surface of the cornea, an iris with significantly deepen rugae, congestion, swelling, moderate circumcorneal hyperemia and existing light reaction, a conjunctiva with diffuse crimson color and are difficult to detect individual vessels, a significant swelling with partial eversion of the lids and whitish secretion in the corner of the eye. In a third animal punctate or diffuse areas of opacity with clearly identifiable details of iris on more than three-quarters of the corneal surface were noted.
72 h after administration of the test material in the first animal punctate or diffuse areas of opacity with clearly identifiable details of iris on more than three-quarters of the surface of the cornea, an iris with significantly deepen rugae, congestion, swelling, moderate circumcorneal hyperemia and existing light reaction, a conjunctiva observed with diffuse crimson color and difficult to identify individual vessels, a marked swelling with partial eversion of the lids whitish secretions around the eye were identified. A second animal showed punctate or diffuse areas of opacity with clearly identifiable details of iris on more than three-quarters of the corneal surface, a marked swelling with partial eversion of the lids and whitish secretion in the corner of the eye. In the third punctate or diffuse areas of opacity with clearly identifiable details of iris on more than three-quarters of the corneal surface, hyperemia and existing light reaction, some hyperemia, conjunctival blood vessels, swelling were observed more than normal and whitish secretion in corner of the eye.
After the instillation of fluorescein, the second and third animal showed punctate or diffuse areas of opacity with clearly identifiable details of iris on more than three-quarters of the corneal surface. In the first animal punctate or diffuse areas of opacity with clearly identifiable details of iris on more than half but less than three quarters of the corneal surface were observed.
7 days after the application of the test material the second animal showed punctate or diffuse areas of opacity with clearly identifiable details of iris on more than half but less than three quarters of the corneal surface in one animal were punctate or diffuse areas of opacity with clearly identifiable details of iris on more than one quarter, but less than half the surface of the cornea, conjunctival hyperemia observed some blood vessels and hair loss around the entire eye. One animal showed punctate or diffuse areas of opacity with clearly identifiable details of iris on more than three-quarters of the surface of the cornea, an iris with clear with significantly deepen rugae, congestion, swelling, moderate circumcorneal hyperemia and existing light reaction, a conjunctiva with diffuse crimson color and difficult to identify individual vessels and hair loss around the entire eye.
After the instillation of fluorescein one animal showed punctate or diffuse areas of opacity with clearly identifiable details of iris to about a quarter of the corneal surface. In two further animals punctate or diffuse areas of opacity with clearly identifiable details of iris on more than half but less than three quarters of the corneal surface were observed.
14 days after the application of the test material one animal circular vascularization on a cornea range between 6 and 3 hours, point-like or diffuse opacity range on more than three-quarters of the corneal surface, a hyperemic conjunctival blood vessels, swelling more al normal and white fluffy secretion. In another animal the medial canthus starting circular vascularization on a cornea range between 5 and 12 h and a cloudy and edematous cornea in the rest of the central region, punctate or diffuse areas of opacity on more than half but less than three quarters of the skin surface and some hyperemic conjunctival blood vessels were observed. One further animal was after 14 days free of signs of eye irritation.
After the instillation of fluorescein one animal showed punctate or diffuse areas of opacity on more than a quarter, but observed less than three quarters of the corneal surface and conjunctival hyperemia some blood vessels. A second animal was after 14 days free of signs of eye irritation.
After the instillation of fluorescein animal the first animal showed punctate or diffuse opacity on more al quarter, but less than half of the corneal surface. In a third animal 0 punctate or diffuse areas of opacity on more than half, but few as three quarters of the corneal surface were observed.
21 days after application of the test showed in one animal pannus over the entire corneal surface, white fluffy secret of the conjuctivae as well as point-like and diffuse areas of opacity for more than three-quarters of the corneal surface. The iris could not be seen due to the full vascularization on the cornea. In one animal vascularisation was observed on the cornea and punctate or diffuse opacity on more than half, but observed less than three quarters of the corneal surface.
After the instillation of fluorescein, two animals showed no punctate or diffuse areas of opacity to about a quarter of the corneal surface.

Table 1 Individual and mean scores of conjuctivae, iris and cornea

Animal N.

Time after treatment

Conjuctivae

Iris

Cornea

 

Chemosis

Redness

Lesion

Opacity

1

24 h

2

2

1

1

48 h

2

2

1

1

72 h

2

2

1

1

 

Mean

2

2

1

1

2

24 h

2

2

1

1

48 h

2

2

1

1

72 h

2

2

1

1

 

Mean

2

2

1

1

3

24 h

2

2

1

1

48 h

1

1

1

1

72 h

1

1

1

1

 

Mean

1.33

1.33

1

1

 

Interpretation of results:
other: Eye damage 1, H318. Classification according to Regulation (EC) No. 1272/2008 (CLP/EU GHS).
Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation

In a reliable skin irritation study performed according to OECD TG 404 and in compliance with GLP three female Albino rabbits (Chbb:HM) were exposed to 0.5 g Reaction mass of N-(1-oxooctadecyl)sarcosine and N-hexadecanoyl-N-methylglycine (EC 947-850-7) onto the clipped skin for 4 h via semi-occlusive coverage (Frey-Tox, 2008). Skin reactions were evaluated according to the Draize scoring system 1, 24, 48 and 72 h and 7 and 14 days post-application. No symptoms of systemic toxicity were observed in any animal during the test period and no mortality occurred during the course of the study. In the initial test, the animal showed well defined erythema and very slight edema formation immediately after application of the test substance. In the main test, slight to well-defined erythema formation was observed in all animals resulting in mean erythema scores over 24, 48 and 72 h of 2, 1 and 2, respectively. Erythema were fully reversible within 21 days in 2/3 animals whereas 1/3 animals still showed isolated scales 21 days after application of the test substance. In addition, slight to moderate edema formation was observed in 3/3 animals resulting in mean edema scores over 24, 48 and 72 h of 1.67, 2.3 and 2, respectively being fully reversible within 7 days in 3/3 animals. No further local or systemic effects were observed. Based on the study results and according to EU classification criteria, the test substance is considered to be irritating to the skin.

Eye irritation

In a reliable eye irritation study with Reaction mass of N-(1-oxooctadecyl)sarcosine and N-hexadecanoyl-N-methylglycine (EC 947-850-7) performed according to OECD TG 405 and in compliance with GLP 0.1 g of the neat test material was instilled in the eye of three Albino rabbits (Chbb:HM) (Frey-Tox, 2009). The eyes were examined and the changes were graded according to the Draize scoring system 1, 24, 48 and 72 h and 7, 14 and 21 days post-application. No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred during the course of the study. Iridial irritation (grade 1) was recorded in 3/3 animals within 24 h after application of the test substance, but being fully reversible within 72 h in 1/3 animals and within 7 days in 2/3 animals, respectively. Corneal opacity (grade 1) was recorded in 3/3 animals but only being reversible in 1/3 animals within the 21 day observation whereas the remaining 2/3 animals showed irreversible effects on the cornea. Conjunctival irritation (grade 1 - 2) was observed in 3/3 animals being reversible within 14 days in 1/3 animals and 21 days in the remaining 2/3 animals, respectively. Chemosis (grade 1 - 2) was also noted in 3/3 animals and was reversible within 7 days in 2/3 animals and within 21 days in 1/3 animals, respectively. The mean values for iridial irritation, corneal opacity, conjunctival irritation and chemosis were calculated to be 1, 1, 2 and 2, respectively. Based on the irreversible effects on the cornea and according to EU classification criteria, the test substance is considered to cause severe eye damage.

Justification for classification or non-classification

While the available data on skin irritation / corrosion of Reaction mass of N-(1-oxooctadecyl)sarcosine and N-hexadecanoyl-N-methylglycine (EC 947-850-7) meet the criteria for classification for Skin Irrit. 2 (H315) according to Regulation (EC) No. 1272/2008 (CLP), the available data on eye irritation / severe eye damage meet the criteria for classification for Eye Damage 1 (H318).