Reaction mass of N,N-ethanediylbis- Hexadecanamide, N-[2-[(1-oxohexadecyl)amino]ethyl]-Octadecanamide, Hexadecanoic acid, 12-hydroxystearic acid, reaction products with ethylenediamine, N,N-ethanediylbis-Octadecanamide and Octadecanoic acid, 12-hydroxystearic acid, reaction products with ethylenediamine

Endpoint summary

Description of key information

Skin irritation was tested in a study according to OECD guideline 404 in rabbits. No effects on the skin (erythema and edema) were observed in all animals at any observation time after application.

Licolub FA 6 was tested for its eye irritant properties in 3 New Zealand White rabbits.The study was performed according to OECD Guideline 405. No effects were observed in all animals 24, 48 and 72 hours after application. The only effect report was redness of the conjunctivae (mean score 1) which was fully reversible.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Principles of method if other than guideline:

With the exception that the test item was put on a surgical gauze patch of approximately 4 cm x 4 cm (instead of 6 cm²) to guarantee a good contact and uniform distribution of the test item on the skin, the study procedures described in this report meet or exceed the requirements of the following guidelines:
OECD Guidelines for Testing of Chemicals, Section 4, number 404 "Acute Dermal Irritation / Corrosion", adopted April 24, 2002.
Commission Directive 2004/73/EC, B.4. "Acute Toxicity: Dermal Irritation/Corrosion", April 29, 2004.

GLP compliance:

yes

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 - 15 weeks (male); 13 - 15 weeks (females)
- Housing: Individually in stainless steel cages equipped with feed hoppers and drinking water bowls. Wood blocks (RCC Ltd, Füllinsdorf) and haysticks 4642 (batch no. 77/05, Provimi Kliba AG) were provided for gnawing.
- Diet (e.g. ad libitum): Pelleted standard Provimi Kliba 3418 rabbit maintenance diet ad libitum (batch no. 85/05) provided by Provimi Kliba AG, CH-4303 Kaiseraugst.
- Water (e.g. ad libitum): Community tap water from Füllinsdorf, ad libitum.
- Acclimation period: 24-MAY-2006 to 28-MAY-2006

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

IN-LIFE DATES:
From: 24.05. To: 01.06.2006

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

water

Amount / concentration applied:

0.5 g (per animal) of the test item was weighed as delivered by the Sponsor and then moistened with approximately 0.5 mL of purified water before application. The pH of the test item was measured before the study initiation date. A formulation of a 1 % (w/w) solution was prepared. The pH was found to be 5 - 6. According to Commission Directive 2004/73/EC, B.4. and OECD Guidelines 404, a test item needs not to be tested if the pH-value is less than 2 or greater than 11.5, owing to its predictable corrosive properties.

Duration of treatment / exposure:

Four days before treatment, the left flank was clipped with an electric clipeer, exposing an area of approximately 100 cm² (10 cm x 10 cm). The skin of the animals was examined one day before treatment, and regrown fur of all animals was clipped again. Animals with overt signs of skin injury or marked irritation which may have interfered with the interpretation of the results were not used in the test. On the day of treatment, 0.5 g of the test item was placed on a surgical gauze patch (ca. 4 cm x 4 cm). This gauze patch was applied to the intact skin of the clipped area. The patch was covered with a semi-occlusive dressing. The dressing was wrapped around the abdomen and anchored with tape. The duration of treatment was 4 hours. Then the dressing was removed and the skin was flushed with lukewarm tap water to clean the application site so that any reactions (erythema) were clearly visible at that time.

Observation period:

The skin reaction was assessed at approximately 1, 24, 48 and 72 hours after exposure (removal of the dressing, gauze patch and test item).

Number of animals:

Number of animals per test: 3 (Animals of both sexes were used)
Age of treatment: 14-15 weeks (male); Male No. 58
13-15 weeks (females); Female Nos. 59 and 60

Irritation parameter:

erythema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Irritant / corrosive response data:

IRRITATION:
The test item did not elicit any skin reactions at the application site of any animal at any of the observation times (all scores 0). The individual mean score for erythema/eschar and oedema for each of the three animals was therefore 0.

COLORATION:
No staining produced by the test item of the treated skin was observed.

CORROSION:
Neither alterations of the treated skin were observed nor were corrosive effects evident on the skin.

Other effects:

VIABILITY/MORTALITY/CLINICAL SIGNS:
No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred.

BODY WEIGHTS:
The body weights of all rabbits were considered to be within the normal range of variability.

Interpretation of results:

other: not irritating to rabbit skin

Conclusions:

Based upon the referred classification criteria (Commission Directive 2001/59/EC of August 2001), the test item is considered to be "not irritating" to rabbit skin.

Executive summary:

The primary skin irritation potential of the substance was investigated according to OECD test guideline no. 404. The test item was applied by topical semi-occlusive application of 0.5 g to the intact left flank of each of three young adult New Zealand White rabbits. The duration of treatment was four hours. The scoring of skin reactions was performed 1, 24, 48 and 72 hours after removal of the dressing.

The test item did not elicit any skin reactions at the application site of any animal at any of the observation times (all scores 0). The individual mean score for erythema/eschar and oedema for each of the three animals was therefore 0.

The application of the substance to the skin resulted in no signs of irritation. The test item caused no staining of the treated skin. No corrosive effects were noted on the treated skin of any animal at any of the measuring intervals and no clinical signs were observed.

Based upon the referred classification criteria (Commission Directive 2001/59/EC of August 2001), the test substance is considered to be "not irritating" to rabbit skin.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

GLP compliance:

yes

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: 14-15 weeks (male), 14-15 weeks (females)
- Housing: Individually in stainless steel cages equipped with feed hoppers and drinking water bowls. Wood blocks (RCC Ltd, Füllinsdorf) and haysticks 4642 (batch no. 77/05, Provimi Kliba AG) were provided for gnawing
- Diet (e.g. ad libitum): Pelleted standard Provimi Kliba 3418 rabbit maintenance diet ad libitum (batch no. 8/06) provided by Provimi Kliba AG, CH-4303 Kaiseraugst
- Water (e.g. ad libitum):Community tap water from Füllinsdorf, ad libitum.
- Acclimation period: 01-JUN-2006 to 06-JUN-2006

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

IN-LIFE DATES:
From: 01.06. To: 19.06.2006

Vehicle:

unchanged (no vehicle)

Amount / concentration applied:

0.1 g (per animal) of the test item reduced into a fine powder was weighed and applied undiluted as delivered by the Sponsor.

Duration of treatment / exposure:

TREATMENT:
The eyes of the animals were examined one day prior to test item administration. Animals with overt signs of ocular injury or irritation which may have interfered with the interpretation of the results were not used in the test.
On the day of treatment, 0.1 g of the test item was placed in the conjunctival sac of the left eye of each animal after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second to prevent loss of test item. The right eye remained untreated and served as the reference control. The treated eyes were not rinsed after instillation.

Observation period (in vivo):

The eyes of each animal were examined approximately after 1, 24, 48 and 72 hours as well as 7 and 10 days after administration. Eye examinations were made with a Varta Cliptrix diagnostic-lamp (Roth AG, CH-4153 Reinach/Switzerland).

Number of animals or in vitro replicates:

Number of animals per test: 3 (Animals of both sexes were used)
Age at treatment: 14-15 weeks (male); Male No. 58;
14-15 weeks (females); Females No. 59 and 60

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

fully reversible

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

0.67

Max. score:

3

Reversibility:

not fully reversible within: 10 days

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Irritant / corrosive response data:

IRRITATION:
The mean score was calculated across 3 scoring times (24, 48 and 72 hours after instillation) for each animal for corneal opacity, iris, redness and chemosis of the conjunctivae, separately. The individual mean scores for corneal opacity and iris were 0.00 for all three animals. The individual mean scores for the conjunctivae were 0.00, 1.00 and 1.00 for reddening and 0.00 for chemosis for all three animals, respectively.
No abnormal findings were observed in the cornea of iris of any animal at any of the measurement intervals.
Slight to moderate reddening of the conjunctivae was noted in all animals at the 1-hour reading and persisted in two animals until the 48-hour reading. Slight to moderate reddening of the sclerae was present in all animals at the 1-hour reading and persisted in two animals until the 48-hour reading. No abnormal findings were observed in the treated eye of any animal 10 days after treatment.

COLORATION:
White remnants of the test item were observed in the eye or conjunctival sac of two animals one hour after treatment.

CORROSION:
No corrosion of the cornea was observed at any of the reading times.

BODY WEIGHTS:
The body weights of all rabbits were considered to be within the normal range of variability.

Other effects:

VIABILITY/MORTALITY AND CLINICAL SIGNS:
No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred.

Interpretation of results:

other: not irritating to the eye

Conclusions:

Based upon the referred classification (Commission Directive 2001/59/EC of August 06, 2001), the substance is considered to be "not irritating" to the rabbit eye.

Executive summary:

The primary eye irritation potential of the substance was investigated according to OECD test guideline no. 405. The test item was applied by instillation of 0.1 g into the left eye of each of three young adult New Zealand White rabbits. Scoring of irritation effects was performed 1, 24, 48 and 72 hours, as well as 7 and 10 days after test item instillation.

The mean score was calculated across 3 scoring times (24, 48 and 72 hours after instillation) for each animal for corneal opacity, iris, redness and chemosis of the conjunctivae, separately. The individual mean scores for the conjunctivae were 0.00, 1.00 and 1.00 for reddening and 0.00 for chemosis for all three animals, respectively.

The instillation of the test item into the eye resulted in mild, early-onset and transient ocular changes, such as reddening of the conjunctivae and sclerae, chemosis and discharge. These effects were reversible and were no longer evident 10 days after treatment. No abnormal findings were observed in the cornea or iris of any animal at any of the examinations. No corrosion was observed at any of the measuring intervals. White remnants of the test item and a slight ocular discharge were observed in the eye or conjunctival sac of two animals one hour after treatment. No clinical signs were observed.

Thus, the test item did not induce significant or irreversible damage to the rabbit eye. Based upon the referred classification criteria (Commission Directive 2001/59/EC of August 06, 2001), the substance is considered to be "not irritating" to the rabbit eye.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Endpoint summary “irritation/corrosion”

Skin irritation: 1 key study available: not irritating

Eye irritation: 1 key study available: not irritating

 

There is one reliable study on the skin irritancy potential. Skin irritation was tested in a guideline study according to OECD guideline 404 in 6 New Zealand White rabbits. The animals were exposed to the test substance for 4 hours. No effects on the skin (erythema and edema) were observed in all animals at any observation time after application.

There is one reliable study available on the eye irritancy potential. Licolub FA 6 was tested for its eye irritant properties in 3 New Zealand White. This study was performed according to OECD Guideline 405. No effects were observed in all animals 24, 48 and 72 hours after application with exception of slight reversible reddening of the eyes.

Justification for selection of skin irritation / corrosion endpoint:
Well performed and reported study equivalent to OECD guideline 404.

Justification for selection of eye irritation endpoint:
Well performed and reported study equivalent to OECD guideline 405.

Justification for classification or non-classification

With reference the reported scores Licolub FA 6 has not to be classified as irritant to the skin according to the criteria laid down in the EU Classification Labelling and Packaging Regulation (1272/2008/EC).

With reference the absence of severe effects Licolub FA 6 does not have to be classified as irritant to the eyes according to the criteria laid down in the EU Classification Labelling and Packaging Regulation (1272/2008/EC).