Registration Dossier

Diss Factsheets

Administrative data

Description of key information

Skin irritation:

The skin irritation property of the registration substance, sodium N-lauroyl glutamate, is derived based on the read-across to sodium N-cocoyl glutamate and sodium N-cocoyl glycinate.

The registration substance and the read-across sources are amides of fatty acids and amino acids and can be characterized as "N-fatty acyl amino acids", of which endogeous occurence and metabolism are known. Based on the comparable chemical structure, comparable phys-chem data and expected comparable metabolism, these compound are likely exhibit comparable toxicity profiles.

Sodium N-cocoyl glutamate is investigated for its skin irritation property according to the OECD 404. 0.5 ml of the test material was applied to the rabbit skin semi occusively for 4 hours. Slight erythema was found in two animals out of three and these effect were reverseible. Sodium N-cocoyl glutamate is not skin irritating.

Sodium N-cocoyl glycinate is investigated for its skin irritation property according to the OECD 404. 0.5 ml of the test material was applied to the rabbit skin semi occusively for 4 hours. Well-defined and mild erythea and edema were found in all treated animals and these effects were reversible. Sodium N-cocoyl glycinate is not skin irritating.

Sodium N-cocoyl glycinate was investigated for its skin irritation property according to the OECD 439. 10 mg of test compound was applied topically to the EPISKIN-SM tissue for 15 minutes followed by a 42 hour post incubation period and immediate determination of cytotoxic effects via MTT reduction assay. No significant effect on the cell viability was obtained. Sodium N-cocoyl glycinate is not skin irritating.

Likewise the registration substance, sodium N-lauroyl glutamate is considered as not skin irritating.

Eye irritation:

The eye irritation property of the registration substance, sodium N-lauroyl glutamate, is derived based on the read-across to sodium N-cocoyl glutamate and sodium N-cocoyl glycinate.

The registration substance and the read-across sources are amides of fatty acids and amino acids and can be characterized as "N-fatty acyl amino acids", of which endogeous occurence and metabolism are known. Based on the comparable chemical structure, comparable phys-chem data and expected comparable metabolism, these compound are likely exhibit comparable toxicity profiles.

Sodium N-cocoyl glutamate was investigated for its eye irritation property according to the OECD 405. 0.1 ml of the test material was applied into the eye of rabbits. Three animals were used. In all treated animals clear irritating effects were found and these effects were reversible within 14 days. It is an eye irritant.

Sodium N-cocoyl glycinate was investigated for its eye irritation property according to the Guideline OECD 405. The test material, 25% sodium N-cocoyl glycinate, was applied to eyes of rabbits. Three animals were used. In all treated animals clear irritating effects were found and these effects were reversible within 14 days. It is an eye irritant.

Sodium N-cocoyl glycinate was investigated for its eye irritation property according to the Guideline OECD 437. The test material was consisted of 68% sodium N-cocoyl glycinate and 20% sodium chloride. The obtained results were indicative of severely irritating effect of the test material. However, due to the high salt content, it is not clear whether the obtained results is to be applied specifically for the sodium N-cocoyl glycinate.

Likewise, the registration substance, sodium N-lauroyl glutamate is considered as an eye irritant.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin irritation / corrosion, other
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
other information
Justification for type of information:
Justification is provided in the separate statement.
Reason / purpose for cross-reference:
assessment report
Reason / purpose for cross-reference:
assessment report
Reason / purpose for cross-reference:
assessment report
Interpretation of results:
GHS criteria not met
Conclusions:
The skin irritation property of the registration substance sodium N-lauroyl glutamate is derived based on the read-across to Sodium N-cocoyl glutamate and Sodium N-cocoyl glycinate. No skin irritating property can be reliably derived.
Executive summary:

The skin irritation property of the registration substance, sodium N-lauroyl glutamate, is derived based on the read-across to sodium N-cocoyl glutamate and sodium N-cocoyl glycinate.

The registration substance and the read-across sources are amides of fatty acids and amino acids and can be characterized as "N-fatty acyl amino acids", of which endogeous occurence and metabolism are known. Based on the comparable chemical structure, comparable phys-chem data and expected comparable metabolism, these compound are likely exhibit comparable toxicity profiles.

Sodium N-cocoyl glutamate is investigated for its skin irritation property according to the OECD 404. 0.5 ml of the test material was applied to the rabbit skin semi occusively for 4 hours. Slight erythema was found in two animals out of three and these effect were reverseible. Sodium N-cocoyl glutamate is not skin irritating.

Sodium N-cocoyl glycinate is investigated for its skin irritation property according to the OECD 404. 0.5 ml of the test material was applied to the rabbit skin semi occusively for 4 hours. Well-defined and mild erythea and edema were found in all treated animals and these effects were reversible. Sodium N-cocoyl glycinate is not skin irritating.

Sodium N-cocoyl glycinate was investigated for its skin irritation property according to the OECD 439. 10 mg of test compound was applied topically to the EPISKIN-SM tissue for 15 minutes followed by a 42 hour post incubation period and immediate determination of cytotoxic effects via MTT reduction assay. No significant effect on the cell viability was obtained. Sodium N-cocoyl glycinate is not skin irritating.

Likewise the registration substance, sodium N-lauroyl glutamate is considered as not skin irritating.

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
1989
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
unchanged (no vehicle)
Amount / concentration applied:
0.5 ml
Duration of treatment / exposure:
4h
Observation period:
7 d
Number of animals:
3
Irritation parameter:
erythema score
Basis:
mean
Remarks:
observations in three animals
Time point:
24/48/72 h
Score:
0.56
Max. score:
1
Reversibility:
fully reversible within: 7 days
Irritation parameter:
edema score
Basis:
mean
Remarks:
observations in three animals
Time point:
24/48/72 h
Score:
0
Max. score:
0

Table: Skin irritation scores

Observation Time

Erythema

Oedema

Animal 1

Animal 2

Animal 3

Animal 1

Animal 2

Animal 3

1h

1

0

1

0

0

0

24h

1

0

1

0

0

0

48h

1

0

1

0

0

0

72h

1

0

0

0

0

0

7 day

0

0

0

0

0

0

Mean of 24/48/72h

1

0

0.67

0

0

0

Interpretation of results:
GHS criteria not met
Conclusions:
Sodium N-cocoyl glutamate was investigated for its skin irritation property according to the Guideline OECD 404. It is not irritating.
Executive summary:

Sodium N-cocoyl glutamate was investigated for its skin irritation property according to the OECD 404. 0.5 ml of the test material was semi occusively applied to the clipped skin area of rabbits for 4 hours. Erythema of score 1 was found in two animals (out of three) which disappeard within 7 days. No oedema was found. Sodium N-cocoyl glutamate is not skin irritating.

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
2008
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Reason / purpose for cross-reference:
read-across source
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
- Source: Harlan Netherlands BV
- Age at study initiation: 14 weeks (males, females)
- Weight at study initiation: 2.2 kg (male); 2.5 kg (females)
- Housing: individually in stainless steel cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: yes

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 - 23 °C
- Humidity (%): 30 - 70 %
- Air changes (per hr): 10 - 15 per hour
- Photoperiod (hrs dark / hrs light): 12 hour cycle dark / light

Type of coverage:
semiocclusive
Preparation of test site:
shaved
Vehicle:
unchanged (no vehicle)
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL / animal
- Concentration (if solution): undiluted

Duration of treatment / exposure:
4 hour
Observation period:
14 days
Number of animals:
3
Details on study design:
TEST SITE
- Area of exposure: approx. 10 cm x 10 cm
- % coverage: 100
- Type of wrap if used: surgical gauze patch

REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes
- Time after start of exposure: 4 hours

SCORING SYSTEM: Irritation scoring according to Council Regulation (EC) No 440/2008
Irritation parameter:
erythema score
Basis:
mean
Remarks:
all animals
Time point:
other: 24h / 48h / 72h/
Score:
1.67
Max. score:
2
Reversibility:
fully reversible
Irritation parameter:
erythema score
Basis:
animal #1
Remarks:
mean
Time point:
other: 24h / 48h / 72h
Score:
1.33
Max. score:
2
Reversibility:
fully reversible
Irritation parameter:
erythema score
Basis:
animal #2
Remarks:
mean
Time point:
other: 24h / 48h / 72h
Score:
1.67
Max. score:
2
Reversibility:
fully reversible
Irritation parameter:
erythema score
Basis:
animal #3
Remarks:
mean
Time point:
other: 24h / 48h / 72h
Score:
2
Max. score:
2
Reversibility:
fully reversible
Irritation parameter:
edema score
Basis:
mean
Remarks:
all animals
Time point:
other: 24h / 48h / 72h
Score:
1.11
Max. score:
2
Reversibility:
fully reversible
Irritation parameter:
edema score
Basis:
animal #1
Remarks:
mean
Time point:
other: 24h / 48h / 72h
Score:
1
Max. score:
1
Reversibility:
fully reversible
Irritation parameter:
edema score
Basis:
animal #2
Remarks:
mean
Time point:
other: 24h / 48h / 72h
Score:
1.33
Max. score:
2
Reversibility:
fully reversible
Irritation parameter:
edema score
Basis:
animal #3
Remarks:
mean
Time point:
other: 24h / 48h / 72h
Score:
1
Max. score:
1
Reversibility:
fully reversible
Other effects:
Scaling noted

Table: Skin irritation scores

Observation Time

Erythema

Oedema

Animal 1

Animal 2

Animal 3

Animal 1

Animal 2

Animal 3

1h

2

2

2

1

2

1

24h

2

2

2

1

2

1

48h

1

2

2

1

1

1

72h

1

1

2

1

1

1

7 day

0

1

0

0

0

0

10 day

0

0

0

0

0

0

14 day

0

0

0

0

0

0

Mean of 24/48/72h

1.33

1.67

2

1

1.33

1

Interpretation of results:
GHS criteria not met
Conclusions:
Sodium N-cocoyl glycinate was investigated for its skin irritation property according to the Guideline OECD 404. It is not skin irritating.
Executive summary:

Sodium N-cocoyl glycinate was investigated for its skin irritation property according to the OECD 404. 0.5 ml of the test material was semi occusively applied to the clipped skin area of rabbits for 4 hours. Three animals were treated. Mild erythema and edema was found in all treated animals and these effects disappeard within 14 days. Sodium N-cocoyl glycinate is not skin irritating.

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
2012-02-28 to 2012-03-05
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
other: EU Method B.46 (Skin Irritation)
Qualifier:
according to guideline
Guideline:
other: OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method
GLP compliance:
yes (incl. QA statement)
Test system:
human skin model
Remarks:
EPISKIN-SM (TM) reconstructed human epidermis model (SkinEthic)
Source species:
human
Cell type:
non-transformed keratinocytes
Vehicle:
unchanged (no vehicle)
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
10 mg + 5 µL aqua dest.
Duration of treatment / exposure:
15 min
Duration of post-treatment incubation (if applicable):
42 +/- 1 h
Number of replicates:
three
Irritation / corrosion parameter:
% tissue viability
Value:
112
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation

 

Name

Negative Control

Positive Control

Test Item

Tissue

1

2

3

1

2

3

1

2

3

absolute OD550

1.325

1.338

1.225

0.144

0.135

0.138

1.471

1.394

1.475

1.260

1.311

1.196

0.143

0.131

0.144

1.408

1.386

1.418

blank-corrected OD550

1.282

1.295

1.182

0.102

0.092

0.095

1.428

1.351

1.432

1.217

1.268

1.153

0.100

0.089

0.101

1.365

1.343

1.375

mean OD550of the duplicates (blank-corrected)

1.250

1.282

1.168

0.101

0.090

0.098

1.397

1.347

1.404

 total mean OD550of 3 replicate tissues (blank-corrected)

1.233*

0.096

1.383

SD OD550

0.06

0.01

0.04

relative tissue viabilities [%]

101.4

103.9

94.7

8.2

7.3

8.0

113.3

109.3

113.8

mean tissue viability [%]

100.0

7.8**

112.1

SD tissue viability [%]***

4.8

0.4

2.5

CV [% viability]

4.8

5.6

2.2

*          Corrected mean OD550of the negative control corresponds to 100% absolute tissue viability.

**      mean relative tissue viability of the three positive control tissues is£ 40%

***     the standard deviation (SD) obtained from the three concurrently tested tissues is 18%

Interpretation of results:
GHS criteria not met
Conclusions:
Sodium N-cocoyl glycinate was investigated for its skin irritation property according to the Guideline OECD 439. It is not skin irritating.
Executive summary:

Sodium N-cocoyl glycinate was investigated for its skin irritation property according to the OECD 439. 10 mg of test compound was applied topically to the EPISKIN-SM tissue for 15 minutes followed by a 42 hour post incubation period and immediate determination of cytotoxic effects via MTT reduction assay. No significant effect on the cell viability was obtained. Sodium N-cocoyl glycinate is not skin irritating.

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

Eye irritation

Link to relevant study records

Referenceopen allclose all

Endpoint:
eye irritation, other
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
other information
Justification for type of information:
Justification is provided in the separate statement.
Interpretation of results:
Category 2 (irritating to eyes) based on GHS criteria
Conclusions:
The eye irritation property of the registration substance sodium N-lauryl glutamate is derived based on the read-across to Sodium N-cocoyl glutamate and Sodium N-cocoyl glycinate. Sodium N-lauroyl glutamate is expected to be an eye irritant.
Executive summary:

The eye irritation property of the registration substance, sodium N-lauroyl glutamate, is derived based on the read-across to sodium N-cocoyl glutamate and sodium N-cocoyl glycinate.

The registration substance and the read-across sources are amides of fatty acids and amino acids and can be characterized as "N-fatty acyl amino acids", of which endogeous occurence and metabolism are known. Based on the comparable chemical structure, comparable phys-chem data and expected comparable metabolism, these compound are likely exhibit comparable toxicity profiles.

Sodium N-cocoyl glutamate was investigated for its eye irritation property according to the OECD 405. 0.1 ml of the test material was applied into the eye of rabbits. Three animals were used. In all treated animals clear irritating effects were found and these effects were reversible within 14 days. It is an eye irritant.

Sodium N-cocoyl glycinate was investigated for its eye irritation property according to the Guideline OECD 405. The test material, 25% sodium N-cocoyl glycinate, was applied to eyes of rabbits. Three animals were used. In all treated animals clear irritating effects were found and these effects were reversible within 14 days. It is an eye irritant.

Sodium N-cocoyl glycinate was investigated for its eye irritation property according to the Guideline OECD 437. The test material was consisted of 68% sodium N-cocoyl glycinate and 20% sodium chloride. The obtained results were indicative of severely irritating effect of the test material. However, due to the high salt content, it is not clear whether the obtained results is to be applied specifically for the sodium N-cocoyl glycinate.

Likewise, the registration substance, sodium N-lauroyl glutamate is considered as an eye irritant.

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
1989
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Reason / purpose for cross-reference:
assessment report
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Vehicle:
unchanged (no vehicle)
Amount / concentration applied:
0.1 ml
Duration of treatment / exposure:
single application
Observation period (in vivo):
14 days
Number of animals or in vitro replicates:
3
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
observations in three treated animals
Time point:
24/48/72 h
Score:
1.78
Max. score:
3
Reversibility:
fully reversible within: 14 days
Remarks on result:
positive indication of irritation
Irritation parameter:
iris score
Basis:
mean
Remarks:
observations in three treated animals
Time point:
24/48/72 h
Score:
1
Max. score:
1
Reversibility:
fully reversible within: 14 days
Remarks on result:
positive indication of irritation
Irritation parameter:
conjunctivae score
Basis:
mean
Remarks:
observations in three treated animals
Time point:
24/48/72 h
Score:
2.89
Max. score:
3
Reversibility:
fully reversible within: 14 days
Remarks on result:
positive indication of irritation
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
observation in three treated animals
Time point:
24/48/72 h
Score:
2.44
Max. score:
3
Reversibility:
fully reversible within: 14 days
Remarks on result:
positive indication of irritation

Eye Irritation Scores

Reading time

Animal

Opacity

Iris

Conjunctivae

Redness

Chemosis

1h

1

d

1

2

3

2

d

1

2

2

3

d

1

2

2

24 hours

1

1

1

3

3

2

1

1

3

2

3

1

1

3

3

48 hours

1

2

1

3

2

2

2

1

3

3

3

2

1

3

3

72 hours

1

2

1

2

1

2

3

1

3

2

3

2

1

3

3

7 day

1

0

0

0

0

2

1v

1

2

1

3

0

1

1c

1

14 day

1

0

0

0

0

2

0

0

0c

0

3

0

0

0c

0

d = dulling of the normal lustre of the corneal surface

v = vascularization of the cornea

c = convulted eyelids

Interpretation of results:
Category 2 (irritating to eyes) based on GHS criteria
Conclusions:
Sodium N-cocoyl glutamate was investigated for its eye irritation property according to the Guideline OECD 405. It is eye irritating.
Executive summary:

Sodium N-cocoyl glutamate was investigated for its eye irritation property according to the OECD 405. 0.1 ml of the test material was applied into the eye of rabbits. The obtained mean scores in three treated animals were 1.8 for corneal opacity, 1.0 for iris lesion, 2.9 for redness of conjuctivae and 2.4 for chemosis of conjuctivae. All the effects were reversible within 14 days. Sodium N-cocoyl glutamate is eye irritating.

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2008
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study according to GLP
Reason / purpose for cross-reference:
read-across source
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
- Source: Harlan Netherlands BV
- Age at study initiation: 15 weeks (male), 16 weeks (females)
- Weight at study initiation: 2.4 kg (male), 2.5 - 2.6 kg (females)
- Housing: individually in stainless steel cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: yes

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 - 23 °C
- Humidity (%): 30 - 70 %
- Air changes (per hr): 10 - 15 per hour
- Photoperiod (hrs dark / hrs light): 12 hour dark / light cycle
Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL
- Concentration (if solution): undiluted
Duration of treatment / exposure:
24 hours
Observation period (in vivo):
7 days
Number of animals or in vitro replicates:
3
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes
- Time after start of exposure: 24 hours

SCORING SYSTEM: numerical scoring system according Council Regulation (EC) 440/2008

TOOL USED TO ASSESS SCORE: fluorescein, diagnostic-lamp
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
all animals
Time point:
other: 24h / 48h / 72h
Score:
0
Max. score:
0
Reversibility:
other: no signs of irritation
Irritation parameter:
cornea opacity score
Basis:
animal #1
Remarks:
mean
Time point:
other: 24h / 48h / 72h
Score:
0
Max. score:
0
Reversibility:
other: no signs of irritation
Irritation parameter:
cornea opacity score
Basis:
animal #2
Remarks:
mean
Time point:
other: 24h / 48h / 72h
Score:
0
Max. score:
0
Reversibility:
other: no signs of irritation
Irritation parameter:
cornea opacity score
Basis:
animal #3
Remarks:
mean
Time point:
other: 24h / 48h / 72h
Score:
0
Max. score:
0
Reversibility:
other: no signs of irritation
Irritation parameter:
iris score
Basis:
mean
Remarks:
all animals
Time point:
other: 24h / 48h / 72h
Score:
0
Max. score:
0
Reversibility:
other: no signs of irritation
Irritation parameter:
iris score
Basis:
animal #1
Remarks:
mean
Time point:
other: 24 h / 48 h / 72h
Score:
0
Max. score:
0
Reversibility:
other: no signs of irritation
Irritation parameter:
iris score
Basis:
animal #2
Remarks:
mean
Time point:
other: 24h / 48h / 72h
Score:
0
Max. score:
0
Reversibility:
other: no signs of irritation
Irritation parameter:
iris score
Basis:
animal #3
Remarks:
mean
Time point:
other: 24h / 48h / 72h
Score:
0
Max. score:
0
Reversibility:
other: no signs of irritation
Irritation parameter:
conjunctivae score
Basis:
mean
Remarks:
all animals
Time point:
other: 24h / 48h / 72h
Score:
1.77
Max. score:
2
Reversibility:
fully reversible
Irritation parameter:
conjunctivae score
Basis:
animal #1
Remarks:
mean
Time point:
other: 24h / 48h / 72h
Score:
1.67
Max. score:
2
Reversibility:
fully reversible
Irritation parameter:
conjunctivae score
Basis:
animal #2
Remarks:
mean
Time point:
other: 24h / 48h / 72h
Score:
2
Max. score:
2
Reversibility:
fully reversible
Irritation parameter:
conjunctivae score
Basis:
animal #3
Remarks:
mean
Time point:
other: 24h / 48h / 72h
Score:
1.67
Max. score:
2
Reversibility:
fully reversible
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
all animals
Time point:
other: 24h / 48h / 72h
Score:
1.77
Max. score:
2
Reversibility:
fully reversible
Irritation parameter:
chemosis score
Basis:
animal #1
Remarks:
mean
Time point:
other: 24h / 48h / 72h
Score:
2
Max. score:
2
Reversibility:
fully reversible
Irritation parameter:
chemosis score
Basis:
animal #2
Remarks:
mean
Time point:
other: 24h / 48h / 72h
Score:
1.33
Max. score:
2
Reversibility:
fully reversible
Irritation parameter:
chemosis score
Basis:
animal #3
Remarks:
mean
Time point:
other: 24h / 48h / 72h
Score:
2
Max. score:
2
Reversibility:
fully reversible

Eye Irritation Scores

Reading time

Animal

Opacity

Iris

Conjunctivae

Redness

Chemosis

1h

1

d

1

2

3

2

d

1

2

2

3

d

1

2

2

24 hours

1

1

1

3

3

2

1

1

3

2

3

1

1

3

3

48 hours

1

2

1

3

2

2

2

1

3

3

3

2

1

3

3

72 hours

1

2

1

2

1

2

3

1

3

2

3

2

1

3

3

7 day

1

0

0

0

0

2

1v

1

2

1

3

0

1

1c

1

14 day

1

0

0

0

0

2

0

0

0c

0

3

0

0

0c

0

d = dulling of the normal lustre of the corneal surface

v = vascularization of the cornea

c = convulted eyelids

Interpretation of results:
Category 2 (irritating to eyes) based on GHS criteria
Conclusions:
Sodium N-cocoyl glycinate was investigated for its eye irritation property according to the Guideline OECD 405. It induced adverse effects in eyes that were reversible within 14 days. It is irritating to eyes.
Executive summary:

Sodium N-cocoyl glycinate was investigated for its eye irritation property according to the Guideline OECD 405. The test material, 25% sodium N-cocoyl glycinate, was applied to eyes of rabbits. Three animals were used. In all treated animals clear irritating effects were found and these effects were reversible within 14 days. It is an eye irritant.

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
2012-02-21 to 2012-05-25
Reliability:
3 (not reliable)
Rationale for reliability incl. deficiencies:
unsuitable test system
Remarks:
Comments on the reliability of the obtained results with regard to hazard assessment of sodium cocoyl glycinate: - The test material contained 20% NaCl. - The applied test design did not include the treatment of the test system only with 20% NaCL. - The applied test system (ex-vivo) is a closed system and therefore is likely to be more susceptible to unspecific physical effects such as osmotic effect. Although the applied test system is to be regarded as valid for the test material, it may not valid for sodium cocoyl glycinate.
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
other: OECD Guidline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants), adopted: 7 September 2009
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
Bayerisches Landesamt für Gesundheit und dLebensmittelsicherheit, München, Germany)
Vehicle:
physiological saline
Amount / concentration applied:
The test item was diluted with physiological saline 0.9% NaCl to gain a 20% concentration.
Duration of treatment / exposure:
750 microL of the test item preparation or the control substance was introduced into the anterior chamber (closed-chamber method).
After 4 hours ± 5 minutes incubation at 32 +/- 1 °C either the test substance or the control substance was removed.
Details on study design:
Test System

Preparation of the Corneas:
The assay uses isolated corneas obtained as a by-product from an abattoir from freshly slaughtered animals
(from Attenberger Fleisch GmbH & Co. KG).
On the test day, fresh eyes were collected from the slautherhouse and were transported in HBSS containing Pen/Strep on ice to the laboratories.
Immediately after arrival of the eyes, cornea preparation was initiated.
The eyes were carefully examined for defects and any defective eyes were discarded.
The tissue surrounding the eyeball was carefully pulled away and the cornea was excised leaving a 2 to 3 mm rim of sclera.
The isolated corneas were stored in a petri dish containing HBSS. Before the corneas were mounted in corneal holders
(MC2, Clermont, France) with the endothelial side against the O-ring of the posterior chamber, they had been visually
examined for defects and any defective cornea had been discarded. The anterior chamber was then positioned on top
of the cornea and tightened with screws. The chambers of the corneal holder were then filled with RPMI (without phenol red)
containing 1% FBS and 2 mM L-glutamine (complete RPMI). The posterior chamber was always filled first. The corneas were
incubated for one hour at 32 +/- 1 °C in a water bath.

Calibration of the Opacitometer:
The opacitometer had been switched on 15 min before the calibration procedure was started. Empty cornea holders were
placed into the opacitometer and the readout was adjusted to zero using the “BAL”-turning knob. For calibration the polyester
foil no. 1 was introduced into the test chamber and the readout was adjusted to 75 using the “CAL”-turning knob. To test the
linearity of the measurement, two additional calibration foils, polyester foil no. 2 and polyester foil no. 3, were measured.
For these, the opacitometer was supposed to display 150 and 225, respectively (± 3%). If this had not been the case,
the calibration procedure would have had to be repeated. The calibration procedure was performed before each test
and was documented in the raw data.

Treatment of the Corneas:
After the equilibration period, the medium was removed from both chambers and replaced with fresh Complete RPMI.
An initial opacity measurement was performed on each of the corneas using an opacitometer (MC2, Clermont, France).
Three corneas with opacity readings approximately equivalent to the median opacity of all corneas were selected as
negative-control corneas. The opacity of each cornea was read against an air-filled chamber and recorded. Corneas
that have an initial opacity reading above 7 units were not dosed. The medium was removed from the anterior chamber
and replaced with the test item or control.
750 microL of the test item preparation or the control substance was introduced into the anterior chamber (closed-chamber method).
After 4 hours ± 5 minutes incubation at 32 +/- 1 °C either the test substance or the control substance was removed
and the epithelium washed at least three times with MEM (containing phenol red). Once the medium was free of test substance,
the cornea was finally rinsed with complete RPMI (without phenol red). The anterior chamber was refilled with complete RPMI
and an opacity measurement was performed.
After the opacity measurement the medium was removed from both chambers of the holder. The posterior chamber was
refilled with fresh complete RPMI. 1 mL of a 5 mg/mL sodium fluorescein solution was added to the anterior chamber and
the corneas were incubated for 90 minutes at 32 +/- 1 °C. Then the medium from the posterior chamber was removed and
its optical density at 490 nm (OD490) was determined, using a spectrophotometer.

Test Groups:
3 corneas for the test item
3 corneas as negative controls treated with physiological saline 0.9% NaCl
3 corneas as positive control treated with imidazole 20% in physiological saline 0.9% NaCl
The BCOP assay is considered to be valid if the in vitro score obtained with the positive control falls within the two
standard deviations of the current historical mean.

Evaluation of Results:
The change in opacity for each cornea was calculated by subtracting the initial opacity reading from the final opacity reading.
These values were corrected by subtracting from each the average change in opacity observed for the negative-control corneas.
The mean opacity value for each treatment was calculated by averaging the corrected opacity values of each cornea for a given treatment.
The mean OD490 for the blank wells were calculated. The mean blank OD490 was subtracted from the OD490 of each well (corrected OD490).
Any dilutions that were made to bring the OD490 values into the linear range of the spectrophotometer (OD490 should be less than 1.500),
were taken into account by multiplying the OD490 value of the dilution by the dilution factor. The final-corrected OD490 of the test article
and the positive control were calculated by subtracting the average corrected OD490 of the negative control corneas from the corrected
OD490 value of each treated cornea:
Final-corrected OD490 = (OD490 – mean blank OD490) – average-corrected negative control OD490
The mean OD490 value of each treatment group was calculated by averaging the final corrected OD490 values of the treated corneas for
that treatment condition.
The following formula was used to determine the in vitro score:
In vitro score = mean opacity value + (15 x mean OD490 value)
Irritation parameter:
in vitro irritation score
Value:
242.15
Vehicle controls validity:
valid
Positive controls validity:
valid
Remarks on result:
positive indication of irritation

Table2:  Opacity

Cornea
No.
Test Item Initial
Opacity
Final
Opacity
Change of
Opacity Value
Corrected
Opacity Value
1 Negative 4 5 1  
2 Control 4 4 0  
3   4 5 1  
MV   4.00 4.67 0.67  
4 Positive 5 229 224 223.33
5 Control 5 168 163 162.33
6   4 216 212 211.33
MV   4.67 204.33 199.67 199.00
7 Test Item 3 187 184 183.33
8   3 283 280 279.33
9   3 172 169 168.33
MV   3.00 214.00 211.00 210.33

 

Table3:  Permeability

Cornea
No.
Test Item OD490 Corrected
OD490 Value
1 Negative 0.020  
2 Control 0.016  
3   0.034  
MV   0.023  
4 Positive 2.114 2.091
5 Control 2.006 1.983
6   1.944 1.921
MV   2.021 1.998
7 Test Item 2.146 2.123
8   2.140 2.117
9   2.148 2.125
MV   2.145 2.121


 

Table4:  In VitroIrritation Score

Cornea
No.
Test Item Corrected
Opacity Value
Corrected
OD490 Value
IVIS
1 Negative 1.00 0.020  
2 Control 0.00 0.016  
3   1.00 0.034  
MV   0.67 0.023 1.02
4 Positive 223.33 2.091  
5 Control 162.33 1.983  
6   211.33 1.921  
MV   199.00 1.998 228.97
7 Test Item 183.33 2.123  
8   279.33 2.117  
9   168.33 2.125  
MV   210.33 2.121 242.15


 

Interpretation of results:
study cannot be used for classification
Conclusions:
Sodium N-cocoyl glycinate was investigated for its eye irritation property according to the Guideline OECD 437. The obtained IVIS score was indicative of severely irritating property. However, due to the high salt content of the test material (20% sodium chloride), it is not clear whether the obtained results is to be applied specifically for the sodium N-cocoyl glycinate.
Executive summary:

Sodium N-cocoyl glycinate was investigated for its eye irritation property according to the Guideline OECD 437. The test material was consisted of 68% sodium N-cocoyl glycinate and 20% sodium chloride. Three bovine corneas were treated with 750 µl test material for 4 hours. The mean opacity score of 199 and mean permeability score of 2.12 were obtained, the corresponding mean IVIS score being 242. However, due to the high salt content, it is not clear whether the obtained results is to be applied specifically for the sodium N-cocoyl glycinate.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Additional information

Justification for classification or non-classification

The skin irritation property of the registration substance sodium N-lauroyl glutamate is derived based on the read-across to Sodium N-cocoyl glutamate and Sodium N-cocoyl glycinate. No skin irritating property can be reliably derived for the registration substance and therefore no classification is assigned.

The eye irritation property of the registration substance sodium N-lauryl glutamate is derived based on the read-across to Sodium N-cocoyl glutamate and Sodium N-cocoyl glycinate. Sodium N-lauroyl glutamate is expected to be an eye irritant. It should be classified as Category 2 based on GHS criteria.