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

Endpoint summary

Description of key information

Tallow-diamine dioleate is considered to be irritating to both skin and eyes.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

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

Adequacy of study:

key study

Study period:

28 Jan 1986 - 11 Feb 1986

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study according to OECD 406 method.

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: 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: Abbaye de Bellefontaine (49122 Bégrolles en Mauges, France)
- Age at study initiation: not indicated
- Weight at study initiation: mean weigh 2.6 kg
- Housing: individually housed in polystyrene cages 0.35 x O.55 x 0.32 m.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 +/- 3°C
- Humidity (%): 50 +/- 20%
- Air changes (per hr): not stated: The inconing, non recycled air, was filtered by an absolute filter.
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 28 Jan 1986 To: 11 Feb 1986

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

other: non-exposed left flank served as control

Amount / concentration applied:

0.5 mL on approx. 6 cm2 on right flank animal

Duration of treatment / exposure:

3 animals for 1 hour, and 3 additional animals for 4 hours.
Residual test substance was removed by means of a gauze saturated with sterile pyrogen-free water.

Observation period:

14 days

Number of animals:

3 per exposure duration of 1 hour and 4 hours

Details on study design:

A dose of 0.5 ml was applied to the clipped area 6 cm2 wide of the right flank of each animal. The treated area was then covered with a hydrophilic gauze patch to prevent loss of test substance. The test substance and the gauze patch were held in contact with the skin using a semi-occluslve patch adhesive tape.

Irritation parameter:

erythema score

Basis:

mean

Remarks:

animals 01, 02, 03 (1 hour)

Time point:

other: 24, 48 and 72 hours

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: Skin dryness was shown reversible by day 13

Irritation parameter:

edema score

Basis:

mean

Remarks:

animals 01, 02, 03 (1 hour)

Time point:

other: 24, 48 and 72 hours

Score:

1.1

Max. score:

4

Reversibility:

fully reversible within: 5 days

Irritation parameter:

erythema score

Basis:

mean

Remarks:

animals 04, 05, 06 (4 hours)

Time point:

other: 24, 48 and 72 hours

Score:

2.1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: Skin dryness was reversible within 12 days

Irritation parameter:

edema score

Basis:

mean

Remarks:

animals 04, 05, 06 (4 hours)

Time point:

other: 24, 48 and 72 hours

Score:

1.6

Max. score:

4

Reversibility:

fully reversible within: 8 days

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

mean

Remarks:

animals 04, 05, 06 (4 hours)

Time point:

other: 24, 48 and 72 hours

Score:

3.67

Max. score:

8

Reversibility:

fully reversible within: 8 days

Remarks on result:

other: Max score is theorethical maximum average score (erythema 4 + oedema 4)

Irritant / corrosive response data:

See scoring table.
The reactions were a bit more marked during the observation period in the animals receiving the test substance during four hours than in those receiving the test substance during one hour.
The report concludes that the test substance INIPOL 002 was considered as irritant.

 Evaluation of skin reactions

		Observation time after exposure
		hours				days
	Animal no.	1 hr	24 hr	48 hr	72 hr	4	5	6	7	8	9	10	11	12	13
Erythema	01	0	2	2	2	2	2	1A	1A	0A	0A	0A	0	0	-
	02	1	3	3	3	3	3	2A	1A	0A	0A	0A	0A	0A	0
	03	0	1	1	1	1	1	1A	0A	0A	0A	0A	0	0	-
	04	2	2	3	4	4	4A	3A	2A	1A	0A	0A	0	0	-
	05	1	2	2	1	1	1	1A	0A	-	-	-	-	0	-
	06	2	2	2	1	1	1	0A	0A	0A	0A	0A	0A	0	-
Oedema	01	0	1	1	0	0	0	0	0	0	0	0	0	0	-
	02	0	2	2	1	1	1	0	0	0	0	0	0	0	0
	03	0	1	1	1	0	0	0	0	0	0	0	0	0	-
	04	2	3	3	4	4	4	3	2	1	0	0	0	0	-
	05	1	1	1	0	0	0	0	0	0	-	-	-	0	-
	06	0	1	1	0	0	0	0	0	0	0	0	0	0	-

A: Skin dryness

 - : Animals without dermal reaction were not recorded any more.

Animals no. 01, 02, 03: Contact duration 1 hour

Animals no. 04, 05, 06: Contact duration 4 hours

Interpretation of results:

irritating

Remarks:

Migrated information Criteria used for interpretation of results: expert judgment

Conclusions:

The test substance was considered as irritant.

Executive summary:

The acute dermal irritation of the test substance INIPOL 002 was evaluated after application of 0.5 ml on the skin of six New-Zealand white rabbits divided into two groups of three animals. The test substance was removed after one hour in one group and after four hours in the other group by means of a gauze patch saturated with sterile pyrogen-free water.

 

One hour after removal of the gauze patch, the signs of dermal irritation were very weak in the animals receiving the test substance during one hour, and moderate in the other animals.

After 24 hours, the signs of irritation were similar and moderate in both groups. They were marked in one animal on day 3 and day 4.

On day 4 an erythema associated with a severe oedema was observed in one animal, as well as a moderate to severe erythema in one animal, a well defined erythema in one animal, a very slight erythema in three animals, and a very slight oedema in two animals.

Oedema lasted until day 8 in one animal.

The erythema noted on day 4 persisted on day 6, and then had disappeared on day 8 in three animals and on day 9 in the last two animals.

On day 9 only skin dryness was noted in 5/6 animals and over the days was observed in less animals, and completely cleared on day 13.

To sum up, the reactions were a bit more marked during the observation period in the animals receiving the test substance during four hours than in those receiving the test substance during one hour.

Under our experimental conditions, the test substance INIPOL 002 was considered as irritant.

ECETOC (Technical Report 66) evaluated this report and calculated the Primary Irritation Index for the three animals that were exposed for 4 hours:

PII = ( SUM Erythema 24/48/72hr + SUM Oedema 24/48/72hr)/ (3 x no. of animals) = 3.67

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

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

Adequacy of study:

key study

Study period:

between 21 and 28 August 1984

Reliability:

1 (reliable without restriction)

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Morton Commercial Rabbits, Stansted, Essex
- Age at study initiation: approximately 9 to 11 weeks of age
- Weight at study initiation: 2.1 to 2.3 kg
- Housing: individually housed in metal cages with perforated floors
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: not indicated; The rabbits selected for the study were all acclimated to the laboratory environment.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): approximately 19°C
- Humidity (%): 30-70%.
- Air changes (per hr): Air exchange was maintained at approximately 19 air changes per hour
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: between 21 and 28 August 1984

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

0.1 mL

Duration of treatment / exposure:

Eye were not flushed after installation.

Observation period (in vivo):

Observations post application at 1 hour and after 1, 2, 3, 4 and 7 days

Number of animals or in vitro replicates:

3

Details on study design:

The eyes of each animal were examined prior to instillation of the test substance to ensure that there was no pre-existing corneal damage or conjunctival inflammation.
A 0.1 ml aliquot of EN 455 was placed into the lower everted lid of one eye of each animal.
The eyelids were then gently held together for one second before releasing. The contralateral eye remained untreated and served as a control.

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 24, 48 and 72 hours

Score:

1.7

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: all animals reach avg score ≥1

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 24, 48, 72 hours

Score:

0.2

Max. score:

2

Reversibility:

fully reversible within: 3 days

Remarks on result:

other: None of the animals reach an avg score ≥ 1

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

other: 24, 48, 72 hours

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: All animals reach avg score ≥ 2

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: 24, 48, 72 hours

Score:

2.2

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: All animals reach avg score ≥ 2

Irritant / corrosive response data:

The numerical scores awarded to the ocular reactions elicited by EN 455 are given in the table on ocular reactions.
All three animals gave a "positive" response.
Corneal opacities developed in all three animals.
Temporary iritis was seen in one animal.
A diffuse crimson red colouration of the conjunctivae was observed in all three animals and was accompanied by considerable swelling with partial eversion of the eyelids or with the eyelids about half-closed.
The eyes were normal seven days after instillation.
There were no other lesions.

Ocular reactions:

rabbit (sex)	Region of Eye		One hour	Day after instillation					Result Positive(+) Negative(-)
				1	2	3	4	7
960 (female)	Cornea		0	2	2	2	2	0	+
	Iris		0	0	0	0	0	0
	Conjunctivae	Redness	2	2	2	2	2	0
		Chemosis	2	3	2	2	2	0
961 (female)	Cornea		0	1	1	1	1	0	+
	Iris		0	0	0	0	0	0
	Conjunctivae	Redness	2	2	2	2	2	0
		Chemosis	2	2	2	2	1	0
978 (female)	Cornea		0	2	2	2	2	0	+
	Iris		0	1	1	0	0	0
	Conjunctivae	Redness	2	2	2	2	2	0
		Chemosis	2	3	2	2	2	0

 

Classification for GHS Cat.2 is fulfilled, as:

- at least 2 of the three animals show an mean (24, 48 and 72 hr) score for corneal opacity ≥ 1 (all three animals),

- at least 2 of the three a mean conjunctival redness of ≥ 2 (all three),

- at least 2 of the three a mean conjunctival chemosis of ≥ 2 (all three)

Interpretation of results:

Category 2A (irritating to eyes) based on GHS criteria

Remarks:

Migrated information

Conclusions:

Oleyl diamine, dioleate was irritating in the in vivo eye irritation test in rabbits. The effects were fully reversible within 7 days. The study results indicate that the substance should be classified Cat.2 irritating to the eyes for GHS.

Executive summary:

The eye irritation potential of Oleyl diamine, dioleate was assessed in an in vivo eye irritation study in rabbits according to OECD 405, and in compliance with GLP.

A 0.1 ml aliquot of Oleyl diamine, dioleate was placed into the lower everted lid of one eye of each animal. The eyelids were then gently held together for one second before releasing. The contralateral eye remained untreated and served as a control.

Examination of the eyes was made after 1 hour and 1, 2, 3, 4, and 7, days after instillation. Observation of the eyes was aided by the use of a handheld torch.

 

Results:

All three animals gave a "positive" response.

Corneal opacities developed in all three animals.

Temporary iritis was seen in one animal.

A diffuse crimson red colouration of the conjunctivae was observed in all three animals and was accompanied by considerable swelling with partial eversion of the eyelids or with the eyelids about half-closed.

The eyes were normal seven days after instillation.

There were no other lesions.

 

According the GHS criteria, these results indicate that the substance should be classified Cat.2 for eye irritation according to GHS.

 

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Dermal irritation:

There is data available on both Tallow-diamine dioleate and Oleyl-diamine dioleate.Cross-reading between the two substances is acceptable on the basis ofidentical chemicalstructures. The only differences between the two substances are related to the alkyl-chains linked to the diamine.Contrary to oleyl do tallow-alkyl chains also contain some C16-chains next to the C18-alkyl chains.The toxicological relevance of this diference is expected to be negligible. The higher level of unsaturation in oleyl-alkyl chainscompared to tallow-alkyl chainscan be considered a worst case representationwith respect to possible irritating properties whencompared to tallow. Due to this very great similarity between these two substances, results obtained from Oleyl-diamine dioleatearefully applicable for the evaluation of Tallow-diamine dioleate as well.

 

There are two studies available on Oleyl-diamine dioleate, both according to OECD 404 guidelines and in compliance to GLP.

 

In one study the acute dermal irritation of Oleyl-diamine dioleate was evaluated after application of 0.5 ml on the skin of 2 groups of 3 NZW rabbits for 1 hour, respectively for 4 hours.

One hour after removal, the signs of dermal irritation were very weak in the animals receiving the test substance during one hour, and moderate in the other animals. During the further observation days, the reactions were a bit more marked during the observation period in the animals receiving the test substance during four hours than in those receiving the test substance during one hour only.

The severest reaction was seen in one animal in the 4-hour group, with a grade 4 reaction for both erythema and oedema 3 and 4, and then gradually decreased and was completely resolved by day 9.

On day 9 only skin dryness was noted in 5/6 animals, and over the days was observed in less animals, and was completely cleared on day 13.

ECETOC evaluated the data and concluded to a Primary Irritation Index of 3.67 for the three animals that were exposed for 4 hours.

The substance was concluded to be irritating to the skin.

 

An additional available study evaluated the skin irritation potential of Oleyl-diamine dioleate, in rabbits. A 0.5 ml aliquot of Oleyl-diamine dioleate was applied under a 2.5 cm square gauze pad to one intact skin site on each of three animals under semi-occlusive dressing for four hours.

At the end of the exposure period, the dressing and pad were removed and the treatment site was washed using distilled water to remove any residual test substance. Observations were done approximately 30 minutes after removal of the patches, and then daily until day 6.

Very slight erythema was observed in all three animals following removal of the bandages, and very slight erythema with slight oedema was seen in all three animals on Days 2 and 3. These reactions gradually ameliorated and the skins were normal by day 5 or 6.

As none of the mean scores for erythema or oedema during three consecutive days was 2.3 or higher, the substance does not need to be classified as skin irritant Cat. 2 for GHS.

 

An older study with Tallow-diamine dioleate resulted to clear irritation, but also indication of possible corrosive effects. However, no observations were performed after 96 hrs, and for these substances it known that they can result to severe inflammatory reactions that often interpreted as corrosive, but which show complete recovery upon extended observations. In view of this, it is noteworthy that for 4 of the 6 animals improvements were already seen at 96 hours compared to 48 hours.

Additionally, the report is of limited validity due to shortcomings in study design and reporting.

 

On overall the available data show mixed results between no need for classification as irritant and possibly even corrosive. The most reliable study of the three concludes to irritant, and this is also considered as the best conclusion based on all available information. Also the available eye irritation study did not show corrosive effects.

 

Additional considerations:

- Definition of corrosion for classification:

A corrosive substance is a substance that produces destruction of skin tissue, namely, visible necrosis through the epidermis and into the dermis, in at least 1 tested animal after exposure up to a 4 hour duration. Corrosive reactions are typified by ulcers, bleeding, bloody scabs and, by the end of observation at 14 days, by discoloration due to blanching of the skin, complete areas of alopecia and scars.

- No real fixed line between very severe irritation and corrosive effects:

 The definition indicates for corrosion effects as "ulcers, bleeding, bloody scabs" should be seen, and by end of 14 day observation "blanching of the skin and scars". However, 'bleeding ulcers' and 'bloody scabs' are rarely observed, and in case very severe reactions indicative for necrosis, the study will be terminated for humane reasons, rather than complete the full observation period.

The study on Tallow-diamine dioleate did not extend observations beyond day 4 because of the severity of the reactions and there is a possibility that recovery could have occurred when the observation was prolonged. However, based on the very severe reactions, the decision to terminate the study was correct in view of animal welfare.

 

- Decontamination of the skin.
These substances are rather sticky to the skin, and will not easily be washed off with pouring some water over it. This could play also a role in view of the observations that reactions can take a longer time to develop. Possibly, cleaning in the studies on Oleyl-diamine dioleate was more efficient. Consequently, it is possible that this difference of substance removal plays a role in the observed difference in severity of reactions.

- Number of animals:
Even in case of testing with three animals, if corrosive effects are observed in one animal, classification is appropriate. In one Oleyl-diamine dioleate study, there is at least one animal that shows severe reactions resulting to the highest scoring, which is the same as the scoring of all three animals in the study with Tallow-diamine dioleate. There were no clear signs of necrosis, although this observation might have been differently evaluated when the other two animals also showed these very severe reactions.

 

 

Eye irritation:

The eye irritation potential of Oleyl-diamine dioleate was assessed in an in vivo eye irritation study in rabbits according to OECD 405, and in compliance with GLP.

A 0.1 ml aliquot of Oleyl diamine, dioleate was placed into the lower everted lid of one eye of each animal. The eyelids were then gently held together for one second before releasing.

Corneal opacities developed in all three animals and temporary iritis was seen in one animal.

A diffuse crimson red colouration of the conjunctivae was observed in all three animals and was accompanied by considerable swelling with partial eversion of the eyelids or with the eyelids about half-closed. The eyes were normal seven days after instillation. There were no other lesions.

 

Respiratory irritation:

Tallow-diamine dioleate is a solid at 20°C with a melting point of 38 °C. It has avapour pressure less than 0.0015 Pa at 20°C (value is an overestimation as it is based on read-across from C12-14-diaminerather than a dioleic acid salt of an alkyl diamine as Tallow-diamine dioleate salt).Also the use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalation route will be unlikely to occur. Consequently, despite the irritant nature of the substance, respiratory irritation is not expected.

Justification for selection of skin irritation / corrosion endpoint:
Most reliable study available.

Justification for selection of eye irritation endpoint:
Only available study

Effects on skin irritation/corrosion: irritating

Effects on eye irritation: irritating

Justification for classification or non-classification

It is proposed to follow the results from the most reliable studies that are available for both Oleyl-diamine dioleate and Tallow-diamine dioleate, and classify for irritation.

Consequently, GHS classification Cat.2 skin irritation is appropriate, with hazard statement H315: Causes skin irritation.

 

In view of the irritant properties, Tallow-diamine dioleate should be classified Cat.2 for eye irritation according to GHS, with hazard statement H319: Causes serious eye irritation.

 

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