Amines, N'-[3-[(3-aminopropyl)amino]propyl]-N,N-di-C16-18-alkyltrimethylenedi-

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

08 June 2016 - 19 July 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

2015

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

May 2008

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

Version / remarks:

August 1998

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No 8147.

Version / remarks:

November 2000, including the most recent partial revisions.

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

date certificate 3 November 2015

Test material

Test material information

Constituent 1

Reference substance name:

[3-({3-[(3-aminopropyl)amino]propyl}amino)propyl](hexadecyl)octadecylamine; [3-({3-[(3-aminopropyl)amino]propyl}amino)propyl]dihexadecylamine; [3-({3-[(3-aminopropyl)amino]propyl}amino)propyl]dioctadecylamine

EC Number:

941-593-4

Cas Number:

1623405-26-4

Molecular formula:

Not applicable UVCB

IUPAC Name:

[3-({3-[(3-aminopropyl)amino]propyl}amino)propyl](hexadecyl)octadecylamine; [3-({3-[(3-aminopropyl)amino]propyl}amino)propyl]dihexadecylamine; [3-({3-[(3-aminopropyl)amino]propyl}amino)propyl]dioctadecylamine

Test material form:

liquid: viscous

Details on test material:

- Name of test material (as cited in study report): di-C16-C18 (evennumbered) alkyl tripropylenetetramine
- Substance type: Viscous liquid
- Physical state: Liquid
- Storage condition of test material: At room temperature container flushed with nitrogen
- Batch/Lot number: B1; batch inspection lot no.:890000394200
- Expiration date of the lot/batch: 22 October 2017
- Purity: 100% (UVCB substance)
- pH: 8.5-10.5 (1% solution)
- specific gravity: 0.83 at 60°C

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

- Source: Charles River France, L’Arbresle Cedex, France.
- Age at study initiation: between 12 and 13 weeks old.
- Weight at study initiation: between 2.351 and 3.211 kg.
- Housing: Animals were housed individually in cages with perforated floors and shelters.
- Diet: Pelleted diet for rabbits (Global Diet 2030 from Harlan Teklad, Italy) approximately 100 grams per day. Hay and wooden sticks were abailable during the study period.
- Water: Free access to tap water.
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS (set to maintain)
- Temperature (°C): 18 – 24
- Humidity (%): 40 - 70
- Air changes (per hr): approx 10
- Photoperiod (hrs dark / hrs light): 12/12

Deviations from the maximum level of daily mean relative humidity occurred. These deviations were mostly transient and not adversely impact the outcome of the study.

IN-LIFE DATES: From: 08 June 2016 to 19 July 2016

Test system

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

other: Adjacent areas of the untreated skin of each animal served as controls.

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.5 mL

Duration of treatment / exposure:

One hour and 4 hours in sentinel animal and 4 hours in two other animals.

Observation period:

14 days.

Number of animals:

3 males.

Details on study design:

STUDY DESIGN
The study was performed in a stepwise manner and was started by treatment of a single rabbit (sentinel) with a stepwise exposure regime. The two other animals were treated with a single patch applied for 4 hours 18 days later, after considering the degree of skin irritation observed in the first animal.

TEST SUBSTANCE PREPARATION
The test item was applied undiluted 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). To facilitate scoring, treated skin areas were re-clipped at least 3 hours before the observations.

The test substance was applied to two separate skin sites of one flank, using a patch of 2x3 cm.

REMOVAL OF TEST SUBSTANCE
One hour or 4 hours after the application, the dressing was removed and the skin cleaned of residual test substance using tap water, watery ethanol (50% v/v) and watery acetone (50% v/v).

OBSERVATIONS
- Mortality/Viability: Twice daily.
- Toxicity: At least once daily.
- Body Weight: Day of treatment (prior to application) and on the day of the final observation.
- Necropsy: No necropsy for gross macroscopic examination was performed according to study plan.
- Irritation:
In the initially treated animal, the skin reactions of all visible treated sites were assessed immediately after removal of a dressing and approximately 1, 24, 48, 72 hours after the removal of the last dressing and test item. After the 4 hours exposure in two further animals, the skin reactions were assessed approximately 1, 24, 48, 72 hours after the removal of the dressing and test item. For the duration of the skin reactions, further observations were made 7 and 14 days after exposure. The irritation scores and a description of all other (local) effects were recorded.

SCORING SYSTEM:
The irritation was assessed according to the numerical scoring system according to OECD 404.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

animal #1

Remarks:

(mean)

Time point:

24/48/72 h

Score:

3

Max. score:

4

Reversibility:

not fully reversible within: 14 days

Remarks:

; Reduced flexibility, scaliness, bald skin

Remarks on result:

other:

Results 2

Irritation parameter:

erythema score

Basis:

animal #2

Remarks:

(mean)

Time point:

24/48/72 h

Score:

1.3

Max. score:

4

Reversibility:

not fully reversible within: 14 days

Remarks:

; Bald skin

Remarks on result:

other:

Results 3

Irritation parameter:

erythema score

Basis:

animal #3

Remarks:

(mean)

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 14 days

Results 4

Irritation parameter:

edema score

Basis:

animal #1

Remarks:

(mean)

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

not fully reversible within: 14 days

Remarks:

; Very slight oedema

Results 5

Irritation parameter:

edema score

Basis:

animal #2

Remarks:

(mean)

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 14 days

Results 6

Irritation parameter:

edema score

Basis:

animal #3

Remarks:

(mean)

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 14 days

Irritant / corrosive response data:

There was no evidence of a corrosive effect on the skin.
Exposure to 0.5 mL of di-C16-C18 (evennumbered) alkyl tripropylenetetramine for 3 minutes, 1 hour or 4 hours in the sentinel animal, all resulted in severe erythema (max. grade 4) and slight oedema (max. grade 2) in the treated skin areas. Four hour exposures to 0.5 mL of di-C16-C18 (evennumbered) alkyl tripropylenetetramine in further two animals resulted in well defined erythema (max. grade 2) and very slight (grade 1) or slight (grade 2) oedema. Highest grades of erythema and oedema were observed at 72 hours and 7 days after application.
Moreover, reduced flexibility of the skin, scabs, scaliness, fissuring of the skin and/or scattered erythema were noted from 72 hours after application onwards in two animals. This had completely resolved at 14 days after application in one of these animals but did not resolve in the other animal. The third animal showed reduced flexibility of the skin from 24 hours after application onwards and scaliness at 7 days after application which had resolved within 14 days after application. The above mentioned effects resulted in bald skin in two animals at 14 days after application.

Other effects:

Remnants of the test item were present on the skin between Days 1 and 8 for all animals. No signs of systemic toxicity were observed in the animals during the test period and no mortality occurred.

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritant) based on GHS criteria

Conclusions:

In a skin irritation study with rabbits, performed according to OECD/EC test guidelines, di-C16-C18 (evennumbered) was classified as skin irritant (Category 2) based on the skin reactions observed.

Executive summary:

A primary skin irritation/corrosion study with di-C16-C18 (evennumbered) alkyl tripropylenetetramine in the rabbit was performed according to OECD/EC guidelines and in compliance with GLP principles. Initially, one rabbit was exposed to three samples of 0.5 mL of di-C16-C18 (evennumbered) alkyl tripropylenetetramine applied to separate skin-sites on intact, clipped skin using a semiocclusive dressing. The exposure periods were 3 minutes, 1 hour and 4 hours, respectively. Skin reactions were assessed at least once daily on Days 1-4 after treatment and 7 and 14 days after exposure. Based on the absence of very severe skin reactions, two further animals were exposed for 4 hours to 0.5 mL di-C16-C18 (evennumbered) alkyl tripropylenetetramine at a later stage.

Skin reactions were assessed 1, 24, 48 and 72 hours and 7 and 14 days after exposure. The exposures in the sentinel animal (3 minutes, 1 hour oand 4 hours) all resulted in severe erythema (max. grade 4) and slight oedema (max. grade 2) in the treated skin areas. Four hour exposures to 0.5 mL of di-C16-C18 (evennumbered) alkyl tripropylenetetramine in further two animals resulted in well defined erythema (max. grade 2) and very slight (grade 1) or slight (grade 2) oedema. Highest grades of erythema and oedema were observed at 72 hours and 7 days after application. Moreover, reduced flexibility of the skin, scabs, scaliness, fissuring of the skin and/or scattered erythema were noted from 72 hours after application onwards in two animals. The third animal showed reduced flexibility of the skin from 24 hours after application onwards and scaliness at 7 days after application. The skin reactions had completely resolved within 14 days after exposure in one animal, whereas the other two animals still showed bald skin respectively reduced flexibility, scaliness and bald skin after 14 days.

Based the skin reactions of the first animal and on the persistence of the skin reactions:

- according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2015) (including all amendments), di-C16-C18 (evennumbered) alkyl tripropylenetetramine should be classified as: skin irritant (Category 2).

- according to the Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments), di-C16-C18 (evennumbered) alkyl tripropylenetetramine should be classified as Irritant (Category 2) and labeled as H315: Causes skin irritation.