Amines, N-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, diacetates

Endpoint summary

Description of key information

Amines, N-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, diacetates (Amines, N-tallow alkyltrimethylenedi-, diacetates) was tested as a mixture containing 2-butoxyethanol (CAS 111-76-2), which found the test compound to be severely irritant for skin but not corrosive. This is consistent with testing conducted on the analogue substance Amines, N-coco alkyltrimethylenedi-, acetates. These results are coherent with the expectation that due to neutralising pH local of the acetate groups, corrosive effects could be lower compared to diamines.

An in vitro eye irritation study was conducted on Amines, N-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, diacetates (Amines, N-tallow alkyltrimethylenedi-, diacetates), which returned an inconclusive result. Considering the method has a good predictivity for substances to be be classified as damaging to the eye, it can be ruled out that the substance is damaging to the eye. Considering that Amines, N-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, diacetates (Amines, N-tallow alkyltrimethylenedi-, diacetates) is irritant to the skin, it is proposed as a conservative approach to classify the substance as irritant 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:

weight of evidence

Study period:

1997-03-28 to 1997-09-08

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

The assay was performed on a mixture containing 2-butoxyethanol (CAS 111-76-2) and no formal analytical certificate was available as the product was in phase of development at that time.

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

Minor fluctuations of relative humidity recorded outside of the target ranges specified in the protocol were not considered to have an impact on the validity or integrity of the study.

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

see above

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Elevage Cunicole de Val de Selle, 80160 Prouzel, France
- Weight at study initiation: 2.4 +/- 0.2 kg.
- Housing: individually in polystyrene cages (35 cm x 55 cm x 32 cm or 48.2 cm x 58 cm x 36.5 cm)
- Diet (e.g. ad libitum): at libitum (112 C pelletet diet, UAR, Villemoisson-sur-Orge, France)
- Water (e.g. ad libitum): ad libitum drinking water filtered by F.G. Millipore membrane (0.22 micron)
- Acclimation period: at least five days before the beginning of the study

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 +/-3
- Humidity (%): 30 to 70%
- Air changes (per hr): 12 cycles/hour of filtered, non-recycled air.
- Photoperiod (hrs dark / hrs light): 12h/12h

IN-LIFE DATES: From: 01 April 1997 To: 15 April 1997

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

0.5 mL

Duration of treatment / exposure:

4 hours

Observation period:

1, 24, 48, 72h after patch removal; animals were observed daily for reversibility of irritating effects up to day 14

Number of animals:

3

Details on study design:

TEST SITE
- Area of exposure: right flank of the animals
- % coverage: approximately 6 cm2
- Type of wrap if used: dry gauze pad held in contact by means of an adhesive hypoallergenic aerated semi-occlusive dressing and a restraining bandage.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): no


SCORING SYSTEM:
Erythema and eschar formation:
. no erythema ........................................................................................................... 0
. very slight erythema (barely perceptible) .............................................................. 1
. well-defined erythema........................................................................................... 2
. moderate to severe erythema................................................................................. 3
. severe erythema (beet redness) to slight eschar formation (injuries in depth) ...... 4

Oedema formation
. no oedema ............................................................................................................. 0
. very slight oedema (barely perceptible) ................................................................ 1
. slight oedema (edges of area well-defined by definite raising)............................. 2
. moderate oedema (raised approximately 1 millimetre)......................................... 3
. severe oedema (raised more than 1 millimetre and extending beyond area
of exposure)........................................................................................................... 4

Any other lesions was noted.

Irritation parameter:

erythema score

Basis:

mean

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

not fully reversible within: 15 Days in 1/3 animals

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

not fully reversible within: 15 Days in 1/3 animals

Irritant / corrosive response data:

On day 1, a well-defined erythema (grade 2) and a severe oedema (grade 4) were noted in all animals. A severe erythema (grade 4) and a severe oedema (grade 4) were observed in all animals from day 2 up to day 5. A very slight to severe erythema (grades 1 to 4) was then noted up to day 11 (one animal), 14 (one animal) or up to the end of the observation period (day 15, in the remaining animal). A severe oedema (grade 4) was observed in one animal from day 11 up to day 15.
Necrosis was recorded in all animals on days 2 and 3, then crusts were observed in all animals from day 4 up to day 9 or 10. Dryness of the skin was noted in two animals, from day 10 or 12 up to the end of the observation period (day 15).
Mean scores over 24, 48 and 72 hours for individual animal were 4.0, 4.0 and 4.0 for erythema and 4.0, 4.0 and 4.0 for oedema.

Interpretation of results:

Category 2 (irritant) based on GHS criteria

Conclusions:

Amines,N-(C16-18 and C18 unsaturated alkyl) trimethylenedi-,diacetates is considered as a skin irritant cat. 2

Executive summary:

The potential of the test item (38% n-tallow alkyltrimethylene diamines, diacetates, mixed with 25% butyl glycol and 37% water) to induce skin irritation was evaluated in rabbits according to OECD (No. 404, 17th July 1992) and EC (92/69/EEC, B.4, 31st July 1992) guidelines. The study was conducted in compliance with the principles of Good Laboratory Practice Regulations.

 

The test item was applied for a period of 4 hours to three male New Zealand White rabbits at a dose-volume of 0.5 mL (undiluted) to the closely-clipped skin of one flank. The test item was held in contact with the skin by means of a semi-occlusive dressing.

 

Cutaneous reactions were observed approximately 1 hour, 24, 48 and 72 hours after removal of the dressing and then daily until the end of the observation period (day 15). The mean values of the scores for erythema and oedema were calculated for each animal. On day 1, a well-defined erythema (grade 2) and a severe oedema (grade 4) were noted in all animals. A severe erythema (grade 4) and a severe oedema (grade 4) were observed in all animals from day 2 up to day 5. A very slight to severe erythema (grades 1 to 4) was then noted up to day 11 (one animal), 14 (one animal) or up to the end of the observation period (day 15, in the remaining animal). A severe oedema (grade 4) was observed in one animal from day 11 up to day 15.Necrosis was recorded in all animals on days 2 and 3, then crusts were observed in all animals from day 4 up to day 9 or 10. Dryness of the skin was noted in two animals, from day 10 or 12 up to the end of the observation period (day 15).

Mean scores over 24, 48 and 72 hours for individual animals were 4.0, 4.0 and 4.0 for erythema and 4.0, 4.0 and 4.0 for oedema.

Under these experimental conditions, the test item is considered to be severely irritant for skin when applied topically to rabbits.

Reference 2

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1989-09-26 to 1990-01-04

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: David Percival Ltd., Moston Sandbach, Cheshire, U.K.
- Age at study initiation: twelve to sixteen weeks old
- Weight at study initiation: 2 to 3 kg
- Housing: individually housed in suspended metal cages.
- Diet (e.g. ad libitum): free access to food (Rabbit Diet, Preston Farmers Limited, New Leake, Boston, Lincolnshire, U.K.) was allowed throughout the study.
- Water (e.g. ad libitum): free acess to drinking water
- Acclimation period: at least five days before the beginning of the study

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 13-21
- Humidity (%): 47 - 69
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light):12/12

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

other: The substance was used moistened with 0.5 mL of distilled water. Before that the solid substance was softened slightly before application, by warming to approximately 40°C

Amount / concentration applied:

0.5 g

Duration of treatment / exposure:

4 hours

Observation period:

1 hour, 24, 48 and 72 hours following the removal of the patches.
If necessary additional observations were made on days seven and fourteen to assess the reversibility of the skin reactions.

Number of animals:

6 rabbits (2 males and 4 females)

Details on study design:

TEST SITE
- Area of exposure: back of the animals
- % coverage: approximately 6,25 cm2
- Type of wrap if used: dry gauze pad held in contact by means of an a strip of surgical adhesive tap. To prevent the animals interfering with the patches and kingesting the test material the trunk of each rabbit was wrapped in an elasticated corset.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes with distilled water


SCORING SYSTEM:
Erythema and eschar formation:
. no erythema ........................................................................................................... 0
. very slight erythema (barely perceptible) .............................................................. 1
. well-defined erythema........................................................................................... 2
. moderate to severe erythema................................................................................. 3
. severe erythema (beet redness) to slight eschar formation (injuries in depth) ...... 4

Oedema formation
. no oedema ............................................................................................................. 0
. very slight oedema (barely perceptible) ................................................................ 1
. slight oedema (edges of area well-defined by definite raising)............................. 2
. moderate oedema (raised approximately 1 millimetre)......................................... 3
. severe oedema (raised more than 1 millimetre and extending beyond area
of exposure)........................................................................................................... 4

Any other lesions was noted.

Irritation parameter:

erythema score

Remarks:

(4-hr exposure)

Basis:

mean

Remarks:

6 rabbits

Time point:

24/48/72 h

Score:

1.9

Max. score:

2

Reversibility:

fully reversible within: 14 days

Irritation parameter:

edema score

Remarks:

4 hr exposure

Basis:

mean

Remarks:

6 rabbits

Time point:

24/48/72 h

Score:

1.9

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritant / corrosive response data:

Moderate to severe dermal responses were produced. The reactions were identified as well-defined erythema, well-defined to moderate oedema, a brown discolouration of the epidermis, scabbing, a dry straw_coloured crust (possible hyperkeratinisation), reduced re-growth of fur, glossy skin and desquamation. No corrosive effects were noted.

Table 1: 4-hour exposure – Individual cutaneous scores recorded for each animal (24, 48 and 72 hours and 7 and 14 days).

 

Skin reaction	Observation time (hr)	Individual scores – Rabbit Number and sex
		Female N° 78	Female N° 82	Female N° 83	Female N° 85	Male N° 86	Male N°94
Erythema and dermal responses not covered by draize scale	1	2	2	2	2	2	2
	24	2	2 Br	2	2 Br	2 Br	2 Br
	48	2 Br	2 Br	2 Br	2 Br	2 Br	2 Br
	72	2 Br	2 Br	2 Br	2 Br	1 Br	2 Br
	7 days	?eStHy	?eSpGFrHy	?eHy	?eHyGFr	0Hy	?eSp
	14 days	0GFr	0GFrD	0 Fr	0FrD	0	0GFrD
Oedema	1	2	2	2	2	2	2
	24	2	3	2	3	3	3
	48	2	2	1	2	2	2
	72	1	2	1	1	1	2
	7 days	0	0	0	0	0	0
	14 days	0	0	0	0	0	0

 

Br = Areas of brown discolouration of the skin surface

D = Desquamation

Fr = reduced re-growth of fur

G = Skin glossy in appearance (keratinolysis)

Hy = Dry, straw-coloured crust, sometimes flaking from the skin (possible hyperkeratinisation)

Sp = Light brown-coloured scab

St = Hardened, dark brown/black-coloured scab

?e = Unable to evaluate degree of erythema due to presence of other adverse dermal responses

Interpretation of results:

Category 2 (irritant) based on GHS criteria

Conclusions:

According to the signs of inflammation still observed at the end of the observation period (14 days) the substance is considered as skin irritant cat.2

Executive summary:

The potential of the test item, Amines, N-coco alkyltrimethylenedi-, acetates (purity unknown) to induce skin irritation was evaluated in rabbits according to OECD (No. 404, 17th July 1992) guideline. The study was conducted in compliance with the principles of Good Laboratory Practice Regulations.

 The test item was applied for a period of 4 hours to six New Zealand White rabbits (four females and two males) at a dose-volume of 0.5 g (moistened with 0.5 mL of distilled water) to the closely-clipped skin of the back. The test item was held in contact with the skin by means of a semi-occlusive dressing.

 Cutaneous reactions were observed approximately 1 hour, 24, 48 and 72 hours after removal of the dressing and then daily and on days 7 and 14.Mean scores over 24, 48 and 72 hours for individual animals were 2-2-2-2-1.7-2 for erythema and 1.7-2.3-1.3-2-2-2.3 for oedema.

Moderate to severe dermal responses were produced. The reactions were identified as well-defined erythema, well-defined to moderate oedema. At the end of the observation period, brown discolouration of the epidermis, scabbing, dry straw-coloured crust (possible hyperkeratinisation), reduced re-growth of fur, glossy skin and desquamation were still observed while erythema and oedema were fully reversible. No corrosive effects were noted.

 

Under these experimental conditions, the test item is considered as a skin irritant when applied topically to rabbits.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

26 April 2012 - 11 May 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

other: OECD guideline 437 (In vitro eye irritation)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

cattle

Strain:

not specified

Details on test animals or tissues and environmental conditions:

Origin: bovine eyes were obtained from freshly slaughtered cattle at the abattoir (SOCAVIA, Cany Barville - France, SOCAVIA, Beuvillers - France or EVA, Saint-Pierre-sur-Dives - France).
Age: bovine cattle were up to 12 months old.
Reason for choice: bovine corneas are recommended by regulatory authorities for this type of study. They are adapted for the evaluation of potential ocular irritants since they are part of the target organ.
Transport from supplier to CiToxLAB France: the eyes were transported to CiToxLAB France at ambient temperature, immerged in buffered Hanks medium containing an antibiotic [Hank’s Balanced Salts Solution (HBSS) plus penicillin/streptomycin (100 units/100 µg/mL final)]. A container with smooth internal surfaces was used for the transport to avoid damage to the corneas.

Preparation of the corneas: the selection and preparation of corneas was performed as soon as possible. At each step of the preparation procedure, care was taken to avoid touching the corneas in order not to damage them.
Selection: a careful macroscopic examination was performed on all eyes to detect the presence of any defects (opacity, scratches, pigmentation, neovascularization, etc). Any eyes with defects were discarded. The examination was performed under a lamp, using HBSS in order to keep the eyes moistened. Particular attention was paid to the corneas and the eyes were swiveled in order to observe the fringe areas and any scratches directly under the light.
Preparation of the selected corneas: the tissues surrounding the eyeball were carefully pulled away and the cornea, surrounded by approximately 2 to 3 mm of sclera, was dissected out. The isolated corneas were stored in HBSS until all corneas had been prepared.
Washing of the corneas: the corneas were washed for 15 minutes, three times, in HBSS plus penicillin/streptomycin (100 units/100 µg/mL final) at room temperature. The corneas were used immediately or within a maximum of 24 hours.
Storage of the corneas: if the corneas were used immediately, after washing they were stored individually in 12 mL of M199 medium containing 5% dextran, plus penicillin/streptomycin, at +4°C, for a maximum of 24 hours before use.

The prepared corneas were stored and used within 24 hours.
(Pre)Incubation T°C: 32°C
Date of experimental phase: 11 May 2012.

Vehicle:

unchanged (no vehicle)

Controls:

other: in vitro negative and positive controls

Duration of treatment / exposure:

Exposure period of 10 minutes, followed by rinsing.

Observation period (in vivo):

Opacity measurement:
- before treatment
- after 2-hour incubation in water

Permeability measurement:
- after 90-min incubation in water (and other procedures), following the 2nd opacity measurement

Number of animals or in vitro replicates:

Not applicable
Triplicate corneas for each timepoint and tested substance (test item, negative control, positive control)

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Rinsing: the anterior part of the eye was emptied and then rinsed 3 times with cMEM.

NEGATIVE CONTROL:
0.9% NaCl solution.

POSITIVE CONTROL:
The positive control was dependent on the physical nature of the test item:
- for non-surfactant liquids and for surfactants (10-minute treatments): 10% sodium hydroxide solution (10% NaOH),
- for non-surfactant solids (4-hour treatments): 20% imidazole solution in 0.9% NaCl (w/v).

SCORING SYSTEM/TOOL
- Opacity:
Using an opacitometer
The average change in opacity during exposure is determined. It is corrected by subtracting the average negative control value from values in positive control and test item.
- Permeability:
Using a spectrophotometer: optical density (OD) at 490 nm wavelength
The optical density is corrected by subtracting the average negative control value from values in positive control and test item.
- Scoring:
In vitro irritancy score (IVIS) = Corrected Opacity + (15 x Corrected OD)

METHOD
Corneas obtained from freshly slaughtered calves were mounted in corneal holders. Both chambers of the corneal holder were filled with complemented MEM culture media (cMEM) and pre-incubated for 1 hour and 5 minutes ± 5 minutes at 32°C. Three corneas were used for each treated series (test item, positive control and vehicle control). Before the treatment, a first opacity measurement was performed using an opacitometer. The test item was tested at the concentration of 10% (w/v) in the vehicle (0.9% NaCl), in a single experiment using a treatment time of 10 minutes. At the completion of the treatment period, the test item was removed from the front opening of the anterior chamber and the epithelium was rinsed. Thecorneas were then incubated for 2 hours at 32°C before a second opacity measurement was performed. After the second opacity measurement, the medium of the anterior chamber was removed and filled with a fluoresceine solution. The holders were then incubated vertically for 90 minutes at 32°C. At the end of the incubation, the optical density of the solution from the posterior chamber of each holder was measured in order to determine the permeability of the cornea. Then the cornea was observed for opaque spots and other irregularities.

Interpretation: see below

Irritation parameter:

in vitro irritation score

Run / experiment:

mean

Value:

31.6

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

valid

Other effects / acceptance of results:

No notable opaque spots or irregularities were observed on vehicle control corneas following the treatment.
Opacity and fluoresceine fixation were observed on the corneas treated with the test item following treatment.
The In Vitro Irritancy Score (IVIS) was: 31.6.

Interpretation of results:

GHS criteria not met

Conclusions:

An IVIS of 31.6 was obtained. The test item is considered to be non damaging to the eye, but the study alone does not allow to conclude on the classification of the substance.

Executive summary:

The objective of this study was to evaluate the potential corrosive or severe irritant properties of the test item on bovine eyes.

The study was performed according to the guideline OECD 437( in vitro eye irritation) and with the principles of Good Laboratory Practices.

 

Corneas obtained from freshly slaughtered calves were mounted in corneal holders. Both chambers of the corneal holder were filled with complemented MEM culture media (cMEM) and pre-incubated for 1 hour and 5 minutes ± 5 minutes at 32°C. Three corneas were used for each treated series (test item, positive control and vehicle control). Before the treatment, a first opacity measurement was performed using an opacitometer.

The test item was tested at the concentration of 10% (w/v) in the vehicle (0.9% NaCl), in a single experiment using a treatment time of 10 minutes. At the completion of the treatment period, the test item was removed from the front opening of the anterior chamber and the epithelium was rinsed. The corneas were then incubated for 2 hours at 32°C before a second opacity measurement was performed. After the second opacity measurement, the medium of the anterior chamber was removed and filled with a fluoresceine solution. The holders were then incubated vertically for 90 minutes at 32°C. At the end of the incubation, the optical density of the solution from the posterior chamber of each holder was measured in order to determine the permeability of the cornea. Then the cornea was observed for opaque spots and other irregularities.

 

No notable opaque spots or irregularities were observed on vehicle control corneas following the treatment. Opacity and fluoresceine fixation were observed on the corneas treated with the test item following treatment. The In Vitro Irritancy Score (IVIS) was: 31.6.

 

The test item is considered to be non damaging to the eye. The study cannot be used alone to conclude on the hazardous properties of the substance to the eyes.

Endpoint conclusion

Endpoint conclusion:

no study available

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation/corrosion

Available data on Amines, N-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, diacetates and amines, N-coco alkyltrimethylenedi-, acetates were found. Even if the study with the first mentioned compound was performed with a mixture containing 2-butoxyethanol and the second one does not provide information on the formulation, a weight of evidence approach could be applied to consider the diamine acetates compounds as irritant. These results are coherent with the expectation that due to neutralising pH local corrosive effects could be lower compared to diamines.

 

The first study was to assess the potential of Amines, N-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, diacetates at 38%, mixed with 25% 2-butoxyethanol and 37% water to induce skin irritation in rabbits according to OECD (No. 404, 17th July 1992) and EC (92/69/EEC, B.4, 31st July 1992) guidelines. The study was conducted in compliance with the principles of Good Laboratory Practice Regulations.

 The test item was applied for a period of 4 hours to three male New Zealand White rabbits at a dose-volume of 0.5 mL (undiluted) to the closely-clipped skin of one flank. The test item was held in contact with the skin by means of a semi-occlusive dressing.

 Cutaneous reactions were observed approximately 1 hour, 24, 48 and 72 hours after removal of the dressing and then daily until the end of the observation period (day 15). The mean values of the scores for erythema and oedema were calculated for each animal. On day 1, a well-defined erythema (grade 2) and a severe oedema (grade 4) were noted in all animals. A severe erythema (grade 4) and a severe oedema (grade 4) were observed in all animals from day 2 up to day 5. A very slight to severe erythema (grades 1 to 4) was then noted up to day 11 (one animal), 14 (one animal) or up to the end of the observation period (day 15, in the remaining animal). A severe oedema (grade 4) was observed in one animal from day 11 up to day 15.Necrosis was recorded in all animals on days 2 and 3, then crusts were observed in all animals from day 4 up to day 9 or 10. Dryness of the skin was noted in two animals, from day 10 or 12 up to the end of the observation period (day 15).

Mean scores over 24, 48 and 72 hours for individual animals were 4.0, 4.0 and 4.0 for erythema and 4.0, 4.0 and 4.0 for oedema.

 

 

In the second study, the potential of Amines, N-coco alkyltrimethylenedi-, acetates (purity unknown) to induce skin irritation was evaluated in rabbits according to OECD (No. 404, 17th July 1992) guideline. The study was conducted in compliance with the principles of Good Laboratory Practice Regulations. The test item was applied for a period of 4 hours to six New Zealand White rabbits (four females and two males) at a dose-volume of 0.5 g (moistened with 0.5 mL of distilled water) to the closely-clipped skin of the back. The test item was held in contact with the skin by means of a semi-occlusive dressing.

Cutaneous reactions were observed approximately 1 hour, 24, 48 and 72 hours after removal of the dressing and then daily and on days 7 and 14.Mean scores over 24, 48 and 72 hours for individual animals were 2-2-2-2-1.7-2 for erythema and 1.7-2.3-1.3-2-2-2.3 for oedema.

Moderate to severe dermal responses were produced. The reactions were identified as well-defined erythema, well-defined to moderate oedema. At the end of the observation period, brown discolouration of the epidermis, scabbing, dry straw-coloured crust (possible hyperkeratinisation), reduced re-growth of fur, glossy skin and desquamation were still observed while erythema and oedema were fully reversible. No corrosive effects were noted.

 

These data indicate that diamine acetates compounds including Amines, N-(C16 -18 and C18 -unsatd.alkyl)trimethylenedi-,diacetates should be considered severely irritant for skin.

 

Eye irritation

The potential corrosive or severe irritant properties of Amines, N-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, diacetates was evaluated on bovine eyes. The study was performed according to the guideline OECD 437 and with the principles of Good Laboratory Practices.

Corneas obtained from freshly slaughtered calves were mounted in corneal holders. Both chambers of the corneal holder were filled with complemented MEM culture media (cMEM) and pre-incubated for 1 hour and 5 minutes ± 5 minutes at 32°C. Three corneas were used for each treated series (test item, positive control and vehicle control). Before the treatment, a first opacity measurement was performed using an opacitometer. The test item was tested at the concentration of 10% (w/v) in the vehicle (0.9% NaCl), in a single experiment using a treatment time of 10 minutes. Then, the test item was removed from the front opening of the anterior chamber and the epithelium was rinsed. The corneas were then incubated for 2 hours at 32°C before a second opacity measurement was performed. After the second opacity measurement, the medium of the anterior chamber was removed and filled with a fluoresceine solution. The holders were then incubated vertically for 90 minutes at 32°C. At the end of the incubation, the optical density of the solution from the posterior chamber of each holder was measured in order to determine the permeability of the cornea. Then the cornea was observed for opaque spots and other irregularities.

 

No notable opaque spots or irregularities were observed on vehicle control corneas following the treatment. Opacity and fluoresceine fixation were observed on the corneas treated with the test item following treatment. The In VitroIrritancy Score (IVIS) was: 31.6 while le limit for classification was = 55.1.

 

Consequently, Amines, N-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, diacetates is considered to be non corrosive and not severely irritant to the eye. According to the OECD Guideline 437, further testing should be conducted for classification and labelling purposes. However, in view of the severely skin irritant properties of Amines, N-(C16 -18 and C18 -unsatd. alkyl)trimethylenedi-, diacetates, the result of the in vitro eye iritation test, and in order not to perform further animal testing, it was decided to consider the substance as eye irritant by default.

Justification for classification or non-classification

The available studies indicate that Amines, N-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, diacetates should be regarded as a severe irritant but not corrosive. Consequently, a GHS classification Cat.2 skin irritation is appropriate, with hazard statement H315: Causes skin irritation.

According to an eye irritation test (OECD Guideline 437), Amines, N-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, diacetates is considered to be non damaging to the eye. In view of the severely skin irritant properties of the substance, it was decide to classify the substance irritating to eyes Cat. 2, with hazard statement H319: Causes serious eye irritation.

STOT-SE Cat.3 for respiratory irritation is not required. Exposure via the inhalation route will be unlikely to occur. Consequently, despite the irritant nature of the substance, respiratory irritation is not expected.