Calcium bis[(Z)-N-methyl-N-(1-oxo-9-octadecenyl)aminoacetate]

Endpoint summary

Description of key information

No data on skin sensitisation is available for the substance calcium bis[(Z)-N-methyl-N-(1-oxo-9-octadecenyl)aminoacetate]. Read across from sodium N-lauroylsarcosinate and N-methyl-N-[C18-(unsaturated)alkanoyl]glycine is used to complete this endpoint.

Both read across substances, sodium N-lauroylsarcosinate and N-methyl-N-[C18-(unsaturated)alkanoyl]glycine, are considered not skin sensitising.

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

Reference 1

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

3 Jul - 31 Jul 1987

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Justification for type of information:

Further information is included under 'Attached justification' in IUCLID section 13 and 'Cross reference'.

Reason / purpose for cross-reference:

read-across: supporting information

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Deviations:

yes

Remarks:

no reliability check included

GLP compliance:

yes

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

A non-LLNA test is available that was performed prior to the current data requirements, stipulated in Regulation (EC) No 1907/2006. In accordance with the same Regulation, the data was included to avoid unnecessary testing.

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Supplied by David Hall Limited
- Age at study initiation: young adult (less than one year old)
- Weight at study initiation: 358-461 g
- Housing: The test comprised of 20 test group and 20 control group guinea pigs housed 5 to a cage and 6 dose ranging guinea pigs housed in a ninth cage. Each aluminium cage had a grid floor beneath which was a peat moss filled tray.
- Diet: The animals were fed on Special Diet Services FD1 guinea pig diet, supplemented with hay, ad libitum.
- Water: ad libitum
- Acclimation period: at least 9 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-21
- Humidity (%): 58
- Photoperiod (hrs dark / hrs light): 12/12

Route:

intradermal and epicutaneous

Vehicle:

water

Concentration / amount:

Main test:
Induction:
Intradermal: 0.05% (v/v)
Epicutaneous: 5 % (v/v)

Challenge:
5 % (v/v)

Route:

epicutaneous, occlusive

Vehicle:

water

Concentration / amount:

Main test:
Induction:
Intradermal: 0.05% (v/v)
Epicutaneous: 5 % (v/v)

Challenge:
5 % (v/v)

No. of animals per dose:

20 (control and test groups)
2 (preliminary test)

Details on study design:

RANGE FINDING TEST
A preliminary range finding test was carried out to assess suitable concentrations for injection and topical application on 4 previously untreated guinea pigs. Injections were in the shaved scapular region and topical application was on the shaved flanks. Two guinea pigs were subjected to intradermal injections and a further 2 guinea pigs were exposed to topical application of the test substance for 48 h at various concentrations.
Injections - 10%, 5%, 2% and 1% (v/v) in distilled water
Topical application - 10%, 5%, 2% and 1% (v/v) in distilled water
Injection responses were assessed 24, 48 and 72 h after injection. Responses to the topical application were assessed 24, 8 and 72 h after patch removal. Before challenging the test animals, a maximum non-irritant concentration of the test material was determined in a preliminary experiment on 2 dose ranging guinea pigs. The animals were pretreated at the induction phase with Freund's Complete Adjuvant only.
The test material was applied to the shaved flanks of the guinea pigs for 24 h at concentrations of 10%, 5%, 2% and 1% v/v (A.I) in distilled water under the same system used for the topical induction except that the patches were left in position for 24 h.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: Example: 2 (intradermal and epicutaneous, respectively)
- Exposure period: Example: single injection (intradermal) and 48 h (epicutaneous)
- Test groups:
Intradermal (3 pairs of injections):
Injection 1: 0.10 mL Freund's Complete Adjuvant
Injection 2: 0.10 mL test material
Injection 3: test substance 0.05% (v/v) in a 1:1 mixture (v/v) FCA/water
- Epicutaneous: test substance in water. Six days after the injection phase, the injection site of each of the test and control group animals was shaved again and then wetted with 10% aqueous SLS to provoke a mild inflammatory response to enhance the possibility of sensitisation.

- Control group:
Intradermal (3 pairs of injections):
Injection 1: 0.10 mL Freund's Complete Adjuvant
Injection 2: 0.10 mL water
Injection 3: a 1:1 mixture (v/v) FCA/water
- Epicutaneous: test substance in water. Six days after the injection phase, the injection site of each of the test and control group animals was shaved again and then wetted with 10% aqueous SLS to provoke a mild inflammatory response to enhance the possibility of sensitisation.

- Site: 4 x 6 cm across the scapular region
- Frequency of applications: 7 days
- Duration: Days 0-8
- Concentrations:: intradermal 0.05%, epicutaneous 5%.

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: 20 (challenge)
- Exposure period: 24 h
- Test groups: test substance and vehicle
- Control group: test substance and vehicle
- Site: Example: right flank (test substance) and left flank (vehicle)
- Concentrations: 5%
- Evaluation (hr after challenge): 48 and 72 h

Positive control substance(s):

no

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

Induction 0.05 and 5%, Challenge: 5%

No. with + reactions:

0

Total no. in group:

20

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: Induction 0.05 and 5%, Challenge: 5%. No with. + reactions: 0.0. Total no. in groups: 20.0.

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

Challenge: 5%

No. with + reactions:

0

Total no. in group:

20

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: Challenge: 5%. No with. + reactions: 0.0. Total no. in groups: 20.0.

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

Induction 0.05 and 5%, Challenge: 5%

No. with + reactions:

0

Total no. in group:

20

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: Induction 0.05 and 5%, Challenge: 5%. No with. + reactions: 0.0. Total no. in groups: 20.0.

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

Challenge: 5%

No. with + reactions:

0

Total no. in group:

20

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: Challenge: 5%. No with. + reactions: 0.0. Total no. in groups: 20.0.

Reading:

1st reading

Hours after challenge:

24

Group:

positive control

Remarks on result:

not measured/tested

Reading:

2nd reading

Hours after challenge:

48

Group:

positive control

Remarks on result:

not measured/tested

PRELIMINARY STUDY

Dose Ranging for Induction:

Severe irritation was noted at sites injected at concentrations of 10%, 5%, 2% and 1% (v/v) in distilled water. A concentration of 0.05% (v/v) in distilled water was selected for injection at induction.

Slight-moderate irritation was noted after 48 h topical exposure under occlusion at a concentration of 10% (v/v) in distilled water. No irritation was observed at 5 % (v/v) after 24 h. Therefore, a concentration of 5% (v/v) in distilled water was selected for topical application at induction.

Dose Ranging for Challenge:

Slight irritation was noted a concentration of 10% (v/v) in distilled water. A concentration of 5% (v/v) in distilled water was selected for challenge.

MAIN STUDY

Induction:

Moderate erythema was noted in test group animals and slight erythema was noted in control group animals.

Challenge:

No reactions were noted in test and control animals after challenge.

Body Weight:

Bodyweights were recorded at test commencement and on test completion. Body weight gains were within acceptable range.

Clinical Signs:

No clinical signs, except skin reactions induced by treatment, were noted at any time point during the study.

Interpretation of results:

other: CLP/EU GHS criteria not met, no classification required according to Regulation(EC) No. 1272/2008

Conclusions:

CLP: not classified

Reference 2

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

3 Jul - 31 Jul 1987

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Justification for type of information:

Further information is included under 'Attached justification' in IUCLID section 13 and 'Cross reference'.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Deviations:

yes

Remarks:

no reliability check included

GLP compliance:

yes

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

A non-LLNA test is available that was performed prior to the current data requirements, stipulated in Regulation (EC) No 1907/2006. In accordance with the same Regulation, the data was included to avoid unnecessary testing.

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Supplied by David Hall Limited
- Age at study initiation: young adult (less than one year old)
- Weight at study initiation: 358-461 g
- Housing: The test comprised of 20 test group and 20 control group guinea pigs housed 5 to a cage and 6 dose ranging guinea pigs housed in a ninth cage. Each aluminium cage had a grid floor beneath which was a peat moss filled tray.
- Diet: The animals were fed on Special Diet Services FD1 guinea pig diet, supplemented with hay, ad libitum.
- Water: ad libitum
- Acclimation period: at least 9 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-21
- Humidity (%): 58
- Photoperiod (hrs dark / hrs light): 12/12

Route:

intradermal and epicutaneous

Vehicle:

water

Concentration / amount:

Main test:
Induction:
Intradermal: 0.05% (v/v)
Epicutaneous: 5 % (v/v)

Challenge:
5 % (v/v)

Route:

epicutaneous, occlusive

Vehicle:

water

Concentration / amount:

Main test:
Induction:
Intradermal: 0.05% (v/v)
Epicutaneous: 5 % (v/v)

Challenge:
5 % (v/v)

No. of animals per dose:

20 (control and test groups)
2 (preliminary test)

Details on study design:

RANGE FINDING TEST
A preliminary range finding test was carried out to assess suitable concentrations for injection and topical application on 4 previously untreated guinea pigs. Injections were in the shaved scapular region and topical application was on the shaved flanks. Two guinea pigs were subjected to intradermal injections and a further 2 guinea pigs were exposed to topical application of the test substance for 48 h at various concentrations.
Injections - 10%, 5%, 2% and 1% (v/v) in distilled water
Topical application - 10%, 5%, 2% and 1% (v/v) in distilled water
Injection responses were assessed 24, 48 and 72 h after injection. Responses to the topical application were assessed 24, 8 and 72 h after patch removal. Before challenging the test animals, a maximum non-irritant concentration of the test material was determined in a preliminary experiment on 2 dose ranging guinea pigs. The animals were pretreated at the induction phase with Freund's Complete Adjuvant only.
The test material was applied to the shaved flanks of the guinea pigs for 24 h at concentrations of 10%, 5%, 2% and 1% v/v (A.I) in distilled water under the same system used for the topical induction except that the patches were left in position for 24 h.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: Example: 2 (intradermal and epicutaneous, respectively)
- Exposure period: Example: single injection (intradermal) and 48 h (epicutaneous)
- Test groups:
Intradermal (3 pairs of injections):
Injection 1: 0.10 mL Freund's Complete Adjuvant
Injection 2: 0.10 mL test material
Injection 3: test substance 0.05% (v/v) in a 1:1 mixture (v/v) FCA/water
- Epicutaneous: test substance in water. Six days after the injection phase, the injection site of each of the test and control group animals was shaved again and then wetted with 10% aqueous SLS to provoke a mild inflammatory response to enhance the possibility of sensitisation.

- Control group:
Intradermal (3 pairs of injections):
Injection 1: 0.10 mL Freund's Complete Adjuvant
Injection 2: 0.10 mL water
Injection 3: a 1:1 mixture (v/v) FCA/water
- Epicutaneous: test substance in water. Six days after the injection phase, the injection site of each of the test and control group animals was shaved again and then wetted with 10% aqueous SLS to provoke a mild inflammatory response to enhance the possibility of sensitisation.

- Site: 4 x 6 cm across the scapular region
- Frequency of applications: 7 days
- Duration: Days 0-8
- Concentrations:: intradermal 0.05%, epicutaneous 5%.

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: 20 (challenge)
- Exposure period: 24 h
- Test groups: test substance and vehicle
- Control group: test substance and vehicle
- Site: Example: right flank (test substance) and left flank (vehicle)
- Concentrations: 5%
- Evaluation (hr after challenge): 48 and 72 h

Positive control substance(s):

no

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

Induction 0.05 and 5%, Challenge: 5%

No. with + reactions:

0

Total no. in group:

20

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: Induction 0.05 and 5%, Challenge: 5%. No with. + reactions: 0.0. Total no. in groups: 20.0.

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

Challenge: 5%

No. with + reactions:

0

Total no. in group:

20

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: Challenge: 5%. No with. + reactions: 0.0. Total no. in groups: 20.0.

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

Induction 0.05 and 5%, Challenge: 5%

No. with + reactions:

0

Total no. in group:

20

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: Induction 0.05 and 5%, Challenge: 5%. No with. + reactions: 0.0. Total no. in groups: 20.0.

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

Challenge: 5%

No. with + reactions:

0

Total no. in group:

20

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: Challenge: 5%. No with. + reactions: 0.0. Total no. in groups: 20.0.

Reading:

1st reading

Hours after challenge:

24

Group:

positive control

Remarks on result:

not measured/tested

Reading:

2nd reading

Hours after challenge:

48

Group:

positive control

Remarks on result:

not measured/tested

PRELIMINARY STUDY

Dose Ranging for Induction:

Severe irritation was noted at sites injected at concentrations of 10%, 5%, 2% and 1% (v/v) in distilled water. A concentration of 0.05% (v/v) in distilled water was selected for injection at induction.

Slight-moderate irritation was noted after 48 h topical exposure under occlusion at a concentration of 10% (v/v) in distilled water. No irritation was observed at 5 % (v/v) after 24 h. Therefore, a concentration of 5% (v/v) in distilled water was selected for topical application at induction.

Dose Ranging for Challenge:

Slight irritation was noted a concentration of 10% (v/v) in distilled water. A concentration of 5% (v/v) in distilled water was selected for challenge.

MAIN STUDY

Induction:

Moderate erythema was noted in test group animals and slight erythema was noted in control group animals.

Challenge:

No reactions were noted in test and control animals after challenge.

Body Weight:

Bodyweights were recorded at test commencement and on test completion. Body weight gains were within acceptable range.

Clinical Signs:

No clinical signs, except skin reactions induced by treatment, were noted at any time point during the study.

Interpretation of results:

other: CLP/EU GHS criteria not met, no classification required according to Regulation(EC) No. 1272/2008

Conclusions:

CLP: not classified

Reference 3

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

09 February 1981 to 19 March 1981

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Justification for type of information:

Further information is included under 'Attached justification' in IUCLID section 13 and 'Cross reference'.

Reason / purpose for cross-reference:

read-across: supporting information

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

not applicable

GLP compliance:

no

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

A non-LLNA test is available that was performed prior to the current data requirements, stipulated in Regulation (EC) No 1907/2006. In accordance with the same Regulation, the data were included to avoid unnecessary testing.

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: EN 41700.92
- Expiration date of the lot/batch: Not reported


FORM AS APPLIED IN THE TEST (if different from that of starting material) : intradermal: in 20% ethanol/80% saline and saline adjuvant mixture (Batco Adjuvant complete Freund, Difco; adjuvant : saline = 1:1)
epidermal: in vaseline PhH VI

Species:

guinea pig

Strain:

other: Pirbright white

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: bred on permises
- Age at study initiation: approx. 10 weeks old
- Weight at study initiation: 270 - 400 g
- Housing: housed individually in Macrolon cages (type 3)
- Diet (e.g. ad libitum): ad libitum standard guinea-pigs pellets - NAFAG No. 830, Gossau SG, supplemented with fresh carrots.
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 10 days
- Indication of any skin lesions: Not reported

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 +/- 1
- Humidity (%): 50 +/- 10
- Air changes (per hr): Not reported
- Photoperiod (hrs dark / hrs light): 14 h of light per day

No. of animals per dose:

10 males and 10 females

Details on study design:

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2 (intradermal and epicutaneous)
- Exposure period: two injections (intradermal) and 48 h (epicutaneous)
- Test groups:
Intradermal :
Two intradermal injections (0.1 ml per injection) made into the neck of the guinea-pigs:
1) a mixture of adjuvant and saline,
2) with the test compound in saline and
3) with the test compound in adjuvant saline mixture.

Epidermal: One week later test item was incorporated in vaseline and applied on a filterpapier patch to the neck of the animals (occlusive administration for 48 hours).
- Control group: yes, 20 animals
- Site: neck
- Concentrations: 5% for intradermal application,
30% for epidermal application (induction), 3% for epidermal application (challenge)

B. CHALLENGE EXPOSURE
- No. of exposures: Two weeks after the epidermal induction application the
animals were tested on the flank with 3% test item in vaseline
- Day(s) of challenge: 24 h occlusive application
- Exposure period: 24 h
- Test groups: test substance (3% in vaseline)
- Control group: vehicle alone (vaseline)
- Site: flank
- Concentrations: 3% in vaseline
- Evaluation (hr after challenge): 48 h

Positive control substance(s):

no

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

control

No. with + reactions:

0

Total no. in group:

20

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

induction: 5 and 30%; challenge: 3%

No. with + reactions:

5

Total no. in group:

20

Remarks on result:

other: Clinical observations: 3 animals with very slight erythema and 2 animals with well defined erythema.

Reading:

1st reading

Hours after challenge:

24

Group:

positive control

Remarks on result:

not measured/tested

Interpretation of results:

GHS criteria not met

Conclusions:

N-methyl-N-(C18-(unsaturated)alkanoyl)glycine is considered not to be a skin sensitiser.

Executive summary:

The skin sensitising properties of N-methyl-N-(C18-(unsaturated)alkanoyl)glycine were tested in a guinea pig maximisation test similar to OECD Guideline 406 (Maurer, 1981).

Twenty Pirbright white guinea pigs were induced with two intradermal injections (0.1 mL per injection) of the test substance at 5% (in 20% ethanol, 80% saline) using the adjuvant complete Freund and an epidermal application of the test substance at 30% (Vaseline PhH VI) on the injection site. The negative control group was included in the study. Epicutaneous challenge exposure was conducted 14 days after the epidermal induction for 24 h under occlusive conditions. 3% of the test substance in Vaseline was applied on the flank. Evaluation of skin reactions was carried out 24 h after removing the dressing. No positive control substance was included in the study.

No skin reactions were observed in any animal in the negative control group. In the treatment group, three animals with very slight erythema and two animals with well defined erythema were observed (corresponding to 25% positive results).

Based on the available data, N-methyl-N-(C18-(unsaturated)alkanoyl)glycine under the conditions of the study.

 

Based on the available data, the test item does not meet the criteria for classification according to Regulation (EC) No 1272/2008 and is considered not to be a skin sensitiser.

Reference 4

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

09 February 1981 to 19 March 1981

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Justification for type of information:

Further information is included under 'Attached justification' in IUCLID section 13 and 'Cross reference'.

Reason / purpose for cross-reference:

read-across source

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

not applicable

GLP compliance:

no

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

A non-LLNA test is available that was performed prior to the current data requirements, stipulated in Regulation (EC) No 1907/2006. In accordance with the same Regulation, the data were included to avoid unnecessary testing.

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: EN 41700.92
- Expiration date of the lot/batch: Not reported


FORM AS APPLIED IN THE TEST (if different from that of starting material) : intradermal: in 20% ethanol/80% saline and saline adjuvant mixture (Batco Adjuvant complete Freund, Difco; adjuvant : saline = 1:1)
epidermal: in vaseline PhH VI

Species:

guinea pig

Strain:

other: Pirbright white

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: bred on permises
- Age at study initiation: approx. 10 weeks old
- Weight at study initiation: 270 - 400 g
- Housing: housed individually in Macrolon cages (type 3)
- Diet (e.g. ad libitum): ad libitum standard guinea-pigs pellets - NAFAG No. 830, Gossau SG, supplemented with fresh carrots.
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 10 days
- Indication of any skin lesions: Not reported

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 +/- 1
- Humidity (%): 50 +/- 10
- Air changes (per hr): Not reported
- Photoperiod (hrs dark / hrs light): 14 h of light per day

No. of animals per dose:

10 males and 10 females

Details on study design:

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2 (intradermal and epicutaneous)
- Exposure period: two injections (intradermal) and 48 h (epicutaneous)
- Test groups:
Intradermal :
Two intradermal injections (0.1 ml per injection) made into the neck of the guinea-pigs:
1) a mixture of adjuvant and saline,
2) with the test compound in saline and
3) with the test compound in adjuvant saline mixture.

Epidermal: One week later test item was incorporated in vaseline and applied on a filterpapier patch to the neck of the animals (occlusive administration for 48 hours).
- Control group: yes, 20 animals
- Site: neck
- Concentrations: 5% for intradermal application,
30% for epidermal application (induction), 3% for epidermal application (challenge)

B. CHALLENGE EXPOSURE
- No. of exposures: Two weeks after the epidermal induction application the
animals were tested on the flank with 3% test item in vaseline
- Day(s) of challenge: 24 h occlusive application
- Exposure period: 24 h
- Test groups: test substance (3% in vaseline)
- Control group: vehicle alone (vaseline)
- Site: flank
- Concentrations: 3% in vaseline
- Evaluation (hr after challenge): 48 h

Positive control substance(s):

no

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

control

No. with + reactions:

0

Total no. in group:

20

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

induction: 5 and 30%; challenge: 3%

No. with + reactions:

5

Total no. in group:

20

Remarks on result:

other: Clinical observations: 3 animals with very slight erythema and 2 animals with well defined erythema.

Reading:

1st reading

Hours after challenge:

24

Group:

positive control

Remarks on result:

not measured/tested

Interpretation of results:

GHS criteria not met

Conclusions:

N-methyl-N-(C18-(unsaturated)alkanoyl)glycine is considered not to be a skin sensitiser.

Executive summary:

The skin sensitising properties of N-methyl-N-(C18-(unsaturated)alkanoyl)glycine were tested in a guinea pig maximisation test similar to OECD Guideline 406 (Maurer, 1981).

Twenty Pirbright white guinea pigs were induced with two intradermal injections (0.1 mL per injection) of the test substance at 5% (in 20% ethanol, 80% saline) using the adjuvant complete Freund and an epidermal application of the test substance at 30% (Vaseline PhH VI) on the injection site. The negative control group was included in the study. Epicutaneous challenge exposure was conducted 14 days after the epidermal induction for 24 h under occlusive conditions. 3% of the test substance in Vaseline was applied on the flank. Evaluation of skin reactions was carried out 24 h after removing the dressing. No positive control substance was included in the study.

No skin reactions were observed in any animal in the negative control group. In the treatment group, three animals with very slight erythema and two animals with well defined erythema were observed (corresponding to 25% positive results).

Based on the available data, N-methyl-N-(C18-(unsaturated)alkanoyl)glycine under the conditions of the study.

 

Based on the available data, the test item does not meet the criteria for classification according to Regulation (EC) No 1272/2008 and is considered not to be a skin sensitiser.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information:

The skin sensitising properties of sodium N-lauroylsarcosinate were investigated in a Guinea pig maximisation test according to EU method B.6 under GLP conditions (Inversek Research International, 1987).

In the main study, 20 female Dunkin-Hartley guinea pigs were induced on Day 0 with intradermal injections of FCA, 0.1 mL of the test substance alone and 0.05% test substance in FCA/water. Epicutaneous induction was done by application of the test substance at 5% in water on Day 8. The negative control group was treated with FCA, water or water/FCA alone. Six days after the injection phase the injection site of each of the test and control group animals was shaved again and then wetted with 10% aqueous SLS to provoke a mild inflammatory response to enhance the possibility of sensitisation. Epicutaneous challenge exposure was conducted 20 days after the first induction for 24 h under occlusive conditions. Test substance was applied at a concentration of 5% on the right flank, and the vehicle was applied on the left flank, respectively. Evaluation of skin reactions was carried out 48 and 72 h after challenge.

Moderate erythema was noted in test group animals, and slight erythema was noted in control group animals after induction. At challenge, no skin reactions were observed in any test and vehicle control animals after 48 and 72 h. No further clinical signs were seen during the study. There were no changes in body weight in any animal during the study period.

Thus, the available data on skin sensitisation do not provide evidence for sensitising properties of sodium N-lauroylsarcosinate.

The skin sensitising properties of N-methyl-N-(C18-(unsaturated)alkanoyl)glycine were tested in a guinea pig maximisation test similar to OECD Guideline 406 (Maurer, 1981).

Twenty Pirbright white guinea pigs were induced with two intradermal injections (0.1 mL per injection) of the test substance at 5% (in 20% ethanol, 80% saline) using the adjuvant complete Freund and an epidermal application of the test substance at 30% (Vaseline PhH VI) on the injection site. The negative control group was included in the study. Epicutaneous challenge exposure was conducted 14 days after the epidermal induction for 24 h under occlusive conditions. 3% of the test substance in Vaseline was applied on the flank. Evaluation of skin reactions was carried out 24 h after removing the dressing.

No skin reactions were observed in any animal in the negative control group. In the treatment group, three animals with very slight erythema and two animals with well defined erythema were observed (corresponding to 25% positive results).Under the conditions of the study, N-methyl-N-(C18-(unsaturated)alkanoyl)glycine was not considered as skin sensitiser.The test item does not meet the criteria for classification according to Regulation (EC) No 1272/2008 and is considered not to be a skin sensitiser.

Both studies were conducted according to the standard guidelines and therefore the data are considered to be reliable and adequate source studies for this endpoint.

Justification for classification or non-classification

Negative results were produced in the studies with two read across substances, sodium N-lauroylsarcosinate and N-methyl-N-[C18-(unsaturated)alkanoyl]glycine.

Therefore, no classification is considered relevant for this endpoint for calcium bis[(Z)-N-methyl-N-(1-oxo-9-octadecenyl)aminoacetate].