9-Octadecenamide, N-[(1S,2S,3R)-2,3-dihydroxy-1-(hydroxymethyl)heptadecyl]-, (9Z)-

Endpoint summary

Description of key information

Skin irritation / corrosion

- Key study, read-across, source substance Ceramide III, OECD guideline 404 and EU Method B.4, GLP, Reliability 1: not irritating to skin.

- Supporting study, target substance Ceramide IIIB, similar to OECD guideline 404, Reliability 3: not irritating to skin.

- Supporting study, human patch test (described in section 7.10.5): no adverse reactions observed; not irritating to skin.

Eye irritation

- Key study, read-across, source substance Ceramide III, OECD guideline 405 GLP and EU method B.5, GLP, Reliability 1: not irritating to the eye

- Supporting study, target substance Ceramide IIIB, similar to OECD guideline 405, Reliability 3: not irritating to the eye.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1992-01-28 to 1992-01-31

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:

May 12, 1981

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Version / remarks:

September 1984

GLP compliance:

yes

Specific details on test material used for the study:

The test substance was determined not to be miscible with water and static.
The test substance was ground to a powder using a mortar and pestle.
To ensure close contact to the animal’s skin, the powdery test substance was applied using a patch, which was moistened with distilled water immediately before use.

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

- Rabbit, New Zealand White, (SPF-Quality)
- Source: Broekman Institute, Someren, The Netherlands
- Age at study initiation: approx. 13 weeks
- Weight at study initiation: 2311-2833 grams
- Housing: conventionally, singly in stainless steel cages with perforated floors and equipped with an automatic drinking system (ITL, Bergen, The Netherlands)
- diet: ad libitum, standard laboratory rabbit diet (LKK-20, pellet diameter 4 mm, Hope Farms, Woerden, The Netherlands) approx. 100 g per day.
- Water: ad libitum, tap-water diluted with decalcified water
- Acclimation period: at least 5 days before start of treatment under test conditions
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21
- Humidity (%): 55
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.5 g
VEHICLE
- no vehicle; test substance was applied using a moistened patch
NEGATIVE CONTROL
- The contralateral flank was similarly prepared, but without test substance
.

Duration of treatment / exposure:

4 hours

Observation period:

50 min, 24 h, 48 h and 72 h after removal of the dressings and test substance

Number of animals:

3

Details on study design:

TEST SITE
- Area of exposure: approx. 6 cm²
- % coverage: 100 %. Due to the physical properties of the test substance the amount of 0.5 g could not be dosed completely within the area that was covered by the patch (6 cm²) and a small amount was applied outside this area.
- Type of wrap if used: moistened Scotchpak-non-woven patch (2x3 cm, 3M, St. Paul, U.S.) mounted on Micropore tape (3M, St. Paul, U.S.). The dressing was wrapped around the abdomen and secured with an elastic bandage (Coban, 3M, St. Paul, U.S.)
REMOVAL OF TEST SUBSTANCE
- the remaining test substance was removed using a tissue moistened with tap-water and subsequently a dry tissue.
- Time after start of exposure: 4 hours
SCORING SYSTEM: the test results were evaluated according to OECD guideline No. 404, “Acute Dermal Irritation/Corrosion”, adopted 28 July 2015.
 

Irritation parameter:

erythema score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal: #1,#2, #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Basis:

animal: #1, #2, #3

Time point:

other: 50 min

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal: #1, #2, #3

Time point:

other: 50 min

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Other effects:

No other effects reported.

IRRITATION

No skin irritation was caused by the test substance after a 4 h dermal application.

CORROSION

There was no evidence of a corrosive effect on the skin.

COLOURATION

No staining of the treated skin by the test substance was observed.

TOXICITY SYMPTOMS, MORTALITY

No symptoms of systemic toxicity were observed in the animals during the test period. No mortality occurred.

Table: Irritant/corrosive response data at each observation time point (mean of all animals).

Score at time point / Reversibility	Erythema	Edema
	Max. score: 4	Max. score: 4
50 min	0/0/0	0/0/0
24 h	0/0/0	0/0/0
48 h	0/0/0	0/0/0
72 h	0/0/0	0/0/0
Average 24h, 48h, 72h	0/0/0	0/0/0

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of this study, the test substance did not cause any irritation and thus, classification is not required.

Executive summary:

In this skin irritation / corrosion study according to OECD Guideline No. 404 (May 12, 1981) and EEC Directive 84/449/EEC, Part B.4 (September 1984), 0.5 g of Ceramide III was applied onto shaved skin of three New Zealand White male albino rabbits using semi-occlusive dressings for 4 hours. An adjacent area of untreated skin, similarly prepared but without test substance and vehicle served as control area.

After 4 hours, Ceramide III was removed using tissue moistened with tap-water. Observations were made after 50 minutes, 24, 48, and 72 h after patch removal.

No reactions were seen; mean erythema and oedema scores after 24, 48, and 72 h were 0 in all animals.

Based on the results of this test, Ceramide III is not irritating to skin.

Reference 2

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

Between 1995-01-03 and 1995-01-19

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

significant methodological deficiencies

Remarks:

only basic information given, only one animal was tested

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

screening test with only one animal

GLP compliance:

not specified

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

no information available

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

unchanged (no vehicle)

Remarks:

The test substance was moistened with distilled water.

Controls:

not specified

Amount / concentration applied:

A sample of 0.5 g of the test substance was moistened with distilled water and subsequently applied to the intact skin of a shaved area on the flank of a male albino rabbit .

Duration of treatment / exposure:

4 hours

Observation period:

60 minutes, 24, 48 and 72 hours after patch removal

Number of animals:

1 (male)

Details on study design:

REMOVAL OF TEST SUBSTANCE
- using a tissue moistened with tap-water and subsequently a dry tissue.
- Time after start of exposure: 4 hours
SCORING SYSTEM: according to OECD Guideline 404
The skin area concerned was re-shaved at least 3 hours before the observations, when considered necessary to facilitate scoring.

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

other: 1 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #1

Time point:

other: 1 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Other effects:

No other effects reported.

Table: Irritant/corrosive response data at each observation time point.

Animal No. 1678 (male)
Time after exposure (h)	Erythema	Oedema
1	0	0
24	0	0
48	0	0
72	0	0
Average 24/48/72 h	0	0

Interpretation of results:

study cannot be used for classification

Conclusions:

The test substance resulted in a primary irritation index of 0 (non-irritating) when applied to the intact skin of a single rabbit.

Executive summary:

Skin irritation potential of Ceramide IIIB was investigated in a study similar to OECD 404. 0.5 g of Ceramide IIIB were applied to the shaved skin of a male New Zealand White rabbit (semi-occlusive coverage). Total exposure was 4 h. Thereafter, the test substance was removed using a tissue moistened with tap-water and a dry tissue.

Observations were made after 60 minutes, 24, 48, and 72 h after patch removal.

No reactions were seen; mean erythema and edema scores after 24, 48, and 72 h were 0.

Based on the results of this test, Ceramide IIIB is not irritating to skin.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1994-10-31 to 1994-11-03

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:

February 24, 1987

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Version / remarks:

December 1992

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

- Source: Broekman Institute, Someren, The Netherlands
- Age at study initiation: approx. 8-14 weeks
- Weight at study initiation: 2100 - 2604 g
- Housing: individual housing in labelled cages with perforated floors (Scanbur, Denmark) and equipped with an automatic drinking system (ITL, Bergen, The Netherlands)
- Diet: ad libitum, standard laboratory rabbit diet (LKK-20, pellet diameter 4 mm, Hope Farms, Woerden, The Netherlands) approx. 100 g per day. In addition, hay (BMI Helmond, The Netherlands) was provided once a week.
- Water: ad libitum, tap-water diluted with decalcified water
- Acclimation period: at least 5 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21
- Humidity (%): 50
- Air changes (per hr): 15
- Photoperiod: fluorescent light (120 lux) 12 hours daily

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 27 +-2 mg (a volume of approx. 0.1 ml)
- Concentration: undiluted

Duration of treatment / exposure:

24 hour

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

3

Details on study design:

REMOVAL OF TEST SUBSTANCE
- After 24 hours (immediately after fluorescein examination) both eyes of one animal was rinsed with approx. 50 ml tepid tap water per eye, using a velocity of flow which did not affect the eyes, to remove residual test substance.
SCORING SYSTEM: according to OECD guideline 405
TOOL USED TO ASSESS SCORE: Readings of ocular reactions were made 1 h, 24 h, 48 h, 72 h after treatment. Besides the eyes were examined for potential eye lesions 24 h by means of 2 % fluorescein-dilution.
It was washed out with 50 ml tepid tap-water per eye, using a velocity of flow which did not affect the eyes, to remove residual test substance.

Irritation parameter:

cornea opacity score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Basis:

animal: #1, #2

Time point:

24/48/72 h

Score:

0

Max. score:

3

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

chemosis score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Other effects:

None described.

IRRITATION

Instillation of 27 mg (a volume of approx. 0.1 ml) into one eye of each of the animals resulted in effects in the iris and the conjunctivae.

The iridic irritation (grade 1) had resolved within 24 hours in all three animals. The irritation of the conjunctivae consisted of redness of the conjunctival tissues and had resolved within 24 hours in two animals and within 72 hours after instillation in the third animal. A small amount of discharge was also noted in the latter animal on day 1.

Remnants of the test substance were noted in the treated eyes on day 1 and/or 2.

CORROSION

There was no evidence of ocular corrosion.

COLOURATION

Remnants of the test substance were noted on the outside of the eyelids on day 1 and/or 2.

TOXIC SYMPTOMS / MORTALITY

No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.

The individual scores of each of the three animals and the average score (24, 48, 72 h) are given in the table below.

	CORNEA	IRIS	CONJUNCTIVAE	CHEMOSIS
Max. score	4	2	3	4
Time point (h)
1	0/0/0	1/1/1	0/0/1	0/0/1
24	0/0/0	0/0/0	0/0/2	0/0/0
48	0/0/0	0/0/0	0/0/1	0/0/0
72	0/0/0	0/0/0	0/0/0	0/0/0
Average 24, 48, 72	0	0	1	0

 

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of this in vivo study, the test item was not irritating to the rabbit eye and thus, no classification is required.

Executive summary:

An acute eye irritation/corrosion study with Ceramide III in the rabbit was carried out in accordance with the OECD guideline No. 405, “Acute Eye Irritation/Corrosion” (February 24, 1987) and the EEC Directive 92/69 /EEC, B.5, “Acute Toxicity - Eye irritation” (December 1992).

Instillation of 27 mg of Ceramide III (a volume of approx. 0.1 ml) into an eye of each of three male New Zealand White albino rabbits resulted in effects on the iris and the conjunctivae in all animals. The untreated eye served as concurrent negative control. The irritation had resolved within 24 hours in two animals and within 72 hours after instillation in the third animal. The mean 24-72 h irritation score for all was 0.0 except for conjunctival redness (1.0) in one rabbit, which resolved completely after 72 h as described above.

No signs of systemic intoxication were observed during the study period.

In conclusion, Ceramide III was not irritating to the rabbit eye in this in vivo study.

Reference 2

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

Between 1995-01-03 and 1995-01-19

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

significant methodological deficiencies

Remarks:

only basic information given, only one animal was tested

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

yes

Remarks:

screening test with only one animal

GLP compliance:

not specified

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

no information available

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 51 mg (a volume of approx. 0.1 ml)
- Concentration: undiluted

Duration of treatment / exposure:

24 hour

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

1 male

Details on study design:

REMOVAL OF TEST SUBSTANCE
-no information available

SCORING SYSTEM: Draize Scheme as stipulated by OECD 405

TOOL USED TO ASSESS SCORE: Readings of ocular reactions were made 1 h, 24 h, 48 h, 72 h after treatment.

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Table: Irritant/corrosive response data at each observation time point. Animal No: 1678 (male). Score given for eyelids (A), nictitating membrane (B) and sclera (C). No discharge was observed after 24 h.

	CORNEA	IRIS	CONJUNCTIVAE	CHEMOSIS
Max. score	4	2	3	4
Time point (h)
1	0	1	2 (C)	2 (A, B, C)
24	0	0	2 (C)	1 (A, B, C)
48	0	0	1 (A, B, C)	0
72	0	0	0	0
Average 24, 48, 72	0	0	1	0.33

Interpretation of results:

study cannot be used for classification

Conclusions:

Under the conditions of this experiment, the test substance was not irritating to the rabbit eye.

Executive summary:

In this study similar to OECD guideline 405, instillation of 51 mg of Ceramide IIIB (approximately 0.1 mL) into an eye of a male New Zealand White albino rabbit did not affect cornea or iris of the animal (average 24-72 h score 0). The average conjunctival redness score (24, 48 and 72 hours) of Ceramide IIIB was 1 and the average chemosis score was 0.33. All effects were reversible at least within 72 hours.

Based on the results of this test, Ceramide IIIB is not irritating to the eye.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available