Amines, N-(3- aminopropyl)-N’-[3-(C18 and C18-unsatd. alkyl amino)propyl]trimethylenedi and amines, N-(3-aminopropyl)-N’-(C18 and C18-unsatd. alkyl)trimethylenedi-

Endpoint summary

Description of key information

Available information indicates that Oleyl tripropylenetetramine is corrosive to skin.

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:

02 February 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No 8147, November 2000, including the most recent partial revisions.

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Substance as described in Chapter 1.1

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan, Belton, Leics, England.
- Age at study initiation: at least 6 weeks
- Weight at study initiation: at least 1.0 kg
- Housing: The animal was individually housed in a labeled cage with perforated floor and shelter
- Diet (e.g. ad libitum): Pelleted diet for rabbits (Global Diet 2030 from Harlan Teklad®, Mucedola, Milanese, Italy) approximately 100 grams per day. Hay (TecniLab-BMI BV, Someren, The Netherlands) was provided at least three times a week.
- Water (e.g. ad libitum): Free access to tap water.
- Acclimation period: The acclimatization period was at least 5 days before the start of treatment under laboratory conditions.

A health inspection was performed prior to the commencement of treatment, to ensure that the animal was in a good state of health. Special attention was paid to the skin to be treated, which was intact and free from abnormalities.

Results of analysis for diet (nutrients and contaminants), hay and water were assessed and did not reveal any findings that were considered to have affected the study integrity. All certificates and results of analysis are retained in the NOTOX archives.

Animal specifications (sex, age and body weight) are specified in the attached table.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18.8 - 20.6ºC
- Humidity (%): 44 - 74%
Cleaning procedures in the room might have caused the temporary fluctuations above the optimal maximum level of 70% for relative humidity. Based on laboratory historical data, these fluctuations were considered not to have affected the study integrity.
- Air changes (per hr): approximately 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: 02 February 2010

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 grams

Duration of treatment / exposure:

Single application.

Observation period:

up to 4 hours after the first application when the single treated animal was sacrificed for ethical reasons.

Number of animals:

1 (based on the severe skin reactions, no further animals were exposed to the test substance)

Details on study design:

STUDY DESIGN
The study was performed in a stepwise manner and started with the treatment of one animal (sentinel) with a stepwise exposure regime. Based on the severe skin reactions, no further animals were exposed to the test substance.

TEST SUBSTANCE PREPARATION
The test substance was applied as delivered by the sponsor.

TEST SITE
Approximately 24 hours before treatment, the dorsal fur was clipped with electric clippers, exposing an area of approximately 150 square centimeters (10x15 cm).

REMOVAL OF TEST SUBSTANCE
After each removal of a dressing, the treated skin was cleaned of residual test substance using water and ethanol.

After the final observation, the animal was sacrificed by intra-venous injection of Euthasol® 20% (AST Farma BV, Oudewater, The Netherlands).

OBSERVATIONS
Mortality/Viability: Twice daily.
Toxicity: At least once daily.
Body Weight: Day of treatment (prior to application).
Necropsy: No necropsy was performed.

The skin reactions of all visible treated sites were assessed immediately after removal of a dressing. The irritation scores and a description of all other (local) effects were recorded. Adjacent areas of untreated skin of the animal served as controls.

SCORING SYSTEM:
The irritation was assessed according to the following numerical scoring system. At each observation, the highest scores given were recorded:

Erythema and eschar formation:
0: No erythema
1: Very slight erythema (barely perceptible)
2: Well-defined erythema
3: Moderate to severe erythema
4: Severe erythema (beet redness) *
*. Where signs of necrosis or corrosion (injuries in depth) prevent erythema scoring, the maximum grade for erythema (= 4) was given.

Oedema formation:
0: No oedema
1; Very slight oedema (barely perceptible)
2: Slight oedema (edges of area well-defined by definite raising)
3: Moderate oedema (raised approximately 1 millimeter)
4: Severe oedema (raised more than 1 millimeter and extending beyond the area of exposure)

Irritation parameter:

erythema score

Basis:

animal #1

Remarks:

(maximum score)

Time point:

other: 3 minutes

Score:

1

Max. score:

4

Reversibility:

not fully reversible within: 4 hours

Irritation parameter:

erythema score

Basis:

animal #1

Remarks:

(maximum score)

Time point:

other: 1 hour

Score:

2

Max. score:

4

Reversibility:

not fully reversible within: 3 hours

Irritation parameter:

erythema score

Basis:

animal #1

Remarks:

(maximum score)

Time point:

other: 4 hours

Score:

4

Max. score:

4

Reversibility:

other: Animal sacrificed for ethical reasons immediately after the observation.

Remarks on result:

other: Dark-brown discolouration surrounded by grey discolouration at the edges of the application area, a sign of necrosis. This observation prevented erythema reading, hence the maximum grade for erythema (=4) was given.

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Reversibility:

other: Not possibe to determine due to animal sacrified for ethical reasons after 4 hours exposure

Remarks on result:

not determinable

Irritation parameter:

edema score

Basis:

animal #1

Remarks:

(maximum score)

Time point:

other: 3 minutes, 1 and 4 hours

Score:

0

Max. score:

4

Reversibility:

not fully reversible within: 4 hours

Irritation parameter:

edema score

Basis:

animal #1

Remarks:

(maximum score)

Time point:

24/48/72 h

Reversibility:

other: Not possible to determine due to animal sacrified for ethical reasons after 4 hours exposure.

Remarks on result:

not determinable

Irritant / corrosive response data:

A 3-minute exposure to 0.5 g of Oleyl tripropylenetetramine resulted in very slight erythema at 1 and 4 hours after exposure.

A 1-hour exposure resulted in very slight erythema immediately after exposure and in well defined erythema at 3 hours after exposure.

A 4-hour exposure resulted in dark brown discolouration surrounded by grey discolouration at the edge of the application area (sign of necrosis) immediately after exposure.

Tables containing individual irritation scores are specified in the attached table.

Sticky or dry remnants of the test substance were present on the skin after exposure.

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

Interpretation of results:

Category 1C (corrosive) based on GHS criteria

Remarks:

Migrated information

Conclusions:

Oleyl tripropylenetetramine should be classified as : skin corrosive (Category 1C).

Executive summary:

One rabbit was exposed to three samples of 0.5 grams of Oleyl tripropylenetetramine applied to separate skin-sites on intact, clipped skin using a semi-occlusive dressing. The exposure periods were 3 minutes, 1 hour and 4 hours, respectively. Skin reactions were assessed up to 4 hours after the 3-minute exposure. Based on severe skin reactions, no further animals were exposed to the test substance.

A 3-minute exposure to 0.5 g of Oleyl tripropylenetetramine resulted in very slight erythema at 1 and 4 hours after exposure.

A 1-hour exposure resulted in very slight erythema immediately after exposure and in well defined erythema at 3 hours after exposure.

A 4-hour exposure resulted in dark brown discolouration surrounded by grey discolouration at the edge of the application area (sign of necrosis) immediately after exposure.

Sticky or dry remnants of the test substance were present on the skin after exposure.

The dark brown discolouration surrounded by grey discolouration at the edge of the application area are signs of necrosis. Following this observation, the animal was sacrificed for humane reasons. These severe skin reactions are expected to result in deep and thick scab formation with possible ruptures of the scab, or the scab may drop off exposing scar tissue. Overall, these skin reactions were considered evidence of full thickness destruction of the skin, and hence no further animals were tested.

Based on these results and according to the:

- Globally Harmonized System of Classification and Labeling of Chemicals (GHS) of the United Nations (2007), Oleyl tripropylenetetramine should be classified as : skin corrosive (Category 1C).

- Regulation (EC) No 1272/2008 on classification, labeling and packaging of substances and mixtures, Oleyl tripropylenetetramine should be classified as Corrosive (Category 1C) and labeled as H314: Causes severe skin burns and eye damage.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (corrosive)

Eye irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Oleyl dipropylenetriamine was evaluated in an in vivo dermal irritation/corrosion study in rabbits (NOTOX, 491219, 2010). Exposure of the skin of one animal for 4 hours with oleyl dipropylenetriamine resulted to evidence of full thickness destruction of the skin, and hence no further animals were tested. It was concluded that Oleyl tripropylenetetramine should be classified as skin corrosive (Category 1C).

 

For the evaluation of dermal corrosion/irritation for REACH substances of various categories of Fatty amines have recently been tested. Available data and practical experience indicated that these substances are corrosive to the skin. However, the information was generally incomplete, often inconsistent and of low validity. For support in the dossiers it was considered to perform the recommended in vitro dermal corrosion studies using human epidermal skin constructs to have adequate endpoint coverage. By comparing the more objective results from these studies, they were thought to be useful to demonstrate classification, in the support of the categories, and for inter- and extrapolation for borderline cases.

Unexpectedly, all the studies performed, including with the polyamine products, indicated that these substances are NOT corrosive in these in vitro test systems using human epidermis constructs, showing viability scores after 3 minutes and 1 hour not much different from control. Also for other fatty nitrile derivatives the same results were obtained.

To evaluate the validity of the in vitro test systems for these substance a few further studies were performed to validate the results within vivostudies.

 

In the interest of animal welfare and to minimize any testing likely to produce severe responses in animals, it was decided not to perform furtherin vivocorrosion studies in rabbits to confirm their corrosive properties.

 

Overview available data on dermal corrosion for polyamines (In bold results on Oleyl tripropylene tetramine):

	dipropylene triamine			Tripropylene tetraamine		dipropylene triamine (branched)		Positive control
Based on FA:	Coco	Tallow	Oleyl	Tallow	Oleyl	C12	Tallow
Skin corrosion - viability in vitro:3 min 1 hour	Non-Cor. 58% 22%	Non-Cor. 98% 96%	Non-Cor. 95% 89%	Not possible	Non-Cor. 91% 85%	Corrosive 43% 42%	-	Corrosive 8-9% 6-9%
Skin corrosion -in vivo	-	-	-	Corr.1C	Corr.1C	Corr.1B (3 min)	Corr.1B (3 min)

(In vitro dermal corrosion: Results considered corrosive when viability is below 50% following 3 minutes, or below 15% following 1 hour exposure)

 

None of the substances reached a viability indicating corrosion following 1 hour exposure, even in a case where in vivo results show corrosion following only 3 minutes exposure. Although in that case the 3 -minute exposure showed a viability below 50% thus indicating corrosion.

The mode of action follow from their structure, consisting of an apolar fatty acid chain and a polar end of a primary amine (linked to one or two secondary amine). The structure can disrupt the cytoplasmatic membrane, leading to lyses of the cell content and consequently the death of the cell.

The similar physicochemical properties among the group of polyamines, also suggest similar behaviour and toxicological responses for these substances.

 

Corrosive to eyes is assumed for substances for which dermal corrosion has been established.

Justification for classification or non-classification

There is one study available that evaluated the in vivo dermal irritation/corrosion potential of Oleyl tripropylenetetramine in rabbits. Based on the results, it is concluded that Oleyl tripropylenetetramine should be classified as corrosive (Cat.1C) under GHS.

 

Corrosive to eyes is assumed for substances for which dermal corrosion has been established.

 

There is no information is available following exposure via inhalation. However, with a vapour pressure less than 4.7 x 10-5 Pa at 20°C, potential for inhalation of vapours is limited. 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, and classification STOT-SE Cat.3 for respiratory irritation is not required.