Amines, C36-alkylenedi-

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

04 - 18 June 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study has been performed according to OECD and/or EC guidelines and according to GLP principles.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nohsan, Notification No 8147, April 2011, including the most recent partial revisions.

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Amines, C36-alkylenedi-

EC Number:

273-282-8

EC Name:

Amines, C36-alkylenedi-

Cas Number:

68955-56-6

Molecular formula:

Not applicable UVCB substance

IUPAC Name:

(1E,19E)-10,11-dioctylicosa-1,19-diene-1,20-diamine

Test material form:

other: Slightly viscous amber liquid

Details on test material:

- Name of test material (as cited in study report): Amines, C36-alkylenedi-
- Substance type: Slightly viscous amber liquid
- Physical state: liquid
- Purity: 98.2%
- Lot/batch No.: 0000436391
- Expiration date of the lot/batch: 24 January 2021
- Storage condition of test material: At room temperature in the dark under nitrogen
- pH (1% in water, indicative): 9.9
- Volatile: No
- Test substance handling: Flush container with nitrogen after handling
- Solubility in Water: No

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

- Source: Charles River Deutschland, Kisslegg, Germany
- Age at study initiation: Animal used within the study was at least 6 weeks old
- Weight at study initiation: Body weight was at least 1.0 kg
- Housing: Animal was housed individually in cage with perforated floors.
- Diet: Free access to pelleted diet for rabbits (Global Diet 2030 from Harlan Teklad, Italy). Hay and wooden sticks were abailable during the study period.
- Water: Free access to tap water.
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
Environmental controls for the animal room were set to maintain 18 to 24°C, a relative humidity of 40 to 70%, approximately 15 room air changes/hour, and a 12-hour light/12-hour dark cycle. Temporary deviations from the minimum level of daily mean relative humidity occurred. Laboratory historical data do not indicate an effect of the deviations.

IN-LIFE DATES: From: 04 - 18 June 2012

Test system

Type of coverage:

semiocclusive

Preparation of test site:

shaved

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:

Single application.

Observation period:

14 days.

Number of animals:

one male

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 severity of the skin reactions, no further animals were exposed.

TEST SUBSTANCE PREPARATION
The test substance 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 the skin of one flank, using a Metalline patch of 2x3 cm.

REMOVAL OF TEST SUBSTANCE
Four hours after the application, the dressing was removed and the skin cleaned of residual test substance using water and ethanol.

OBSERVATIONS
- Mortality/Viability: Twice daily.
- Toxicity: At least once daily.
- Body Weight: Day of treatment (prior to application) and after the final observation.
- Necropsy: No necropsy was performed according to protocol.
- Irritation: 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 substance. For the duration of the skin reactions, further observations were made 7 and 14 (maximum)
days after exposure. 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 numerical scoring system according to OECD 404.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

mean

Time point:

other: 24, 48 and 72 hours.

Score:

3

Max. score:

4

Results 2

Irritation parameter:

edema score

Basis:

mean

Time point:

other: 24, 48 and 72 hours.

Score:

3

Max. score:

4

Irritant / corrosive response data:

Irritation: A 3-minute, 1-hour and 4-hour exposure resulted in erythema (maximum grade 4 (severe)) and oedema (maximum grade 2 (slight) to 4 (severe)), which had resolved within 14 days following exposure. Superficial brown discoloration (a sign of necrosis) and superficial fissuring of the skin were noted at 7 days after exposure, and bald skin (bald and shiny after a 1- and 4-hour exposure) and scaliness were noted at 14 days after exposure. At 7 days after a 1- and 4-hour exposure, skin reactions could not be scored due to fissuring and brown discolouration (a sign of necrosis; due to which a maximum grade 4 was given for erythema).

Corrosion: There was no evidence of a corrosive effect on the skin.

Other effects:

Coloration/remnants: On the skin exposed to the test substance for 1 and 4 hours, remnants of the test substance were present, being sticky on Day 1, and 24, 48 and 72 hours after exposure, and dry at 7 days after exposure. This did not hamper the scoring of the skin irritation reactions.

Toxicity/mortality: No symptoms 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)

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

No evidence of full thickness destruction of the skin or scar tissue was observed during the observation period, indicating that no corrosion of the skin had occurred by dermal application of Amines, C36-alkylenedi- to the intact rabbit skin. The extreme response following a 4-hour exposure to Amines, C36-alkylenedi- in a single animal was irreversible within the 14-day observation period.
Based on these results and according to the:
- Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2007), Amines, C36-alkylenedi- should be classified as : skin irritant (Category 2).
- Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures, Amines, C36-alkylenedi- should be classified as Irritant (Category 2) and labeled as H315: Causes skin irritation.

Executive summary:

Primary skin irritation/corrosion study with AMINES, C36-ALKYLENEDI- in the rabbit (semi-occlusive application). The study was carried out based on the guidelines described in:

OECD No.404 (2002); "Acute Toxicity - Skin irritation"

EC, No 440/2008; B4: "Acute Toxicity: Dermal Irritation/Corrosion"

US EPA, OPPTS 870.2500 (1998), Acute Dermal Irritation

JMAFF Guidelines (2000) including the most recent partial revisions.

 

One rabbit was exposed to three samples of 0.5 mL of AMINES, C36-ALKYLENEDI- 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 at least once daily for 4 days after exposure and 7 and 14 days after exposure. Based on the severity of the skin reactions, no further animals were exposed.

 

A 3-minute, 1-hour and 4-hour exposure resulted in erythema (maximum grade 4 (severe)) and oedema (maximum grade 2 (slight) to 4 (severe)), which had resolved within 14 days following exposure. Superficial brown discoloration (a sign of necrosis) and superficial fissuring of the skin were noted at 7 days after exposure, and bald skin (bald and shiny after a 1- and 4-hour exposure) and scaliness were noted at 14 days after exposure.

At 7 days after a 1- and 4-hour exposure, skin reactions could not be scored due to fissuring and brown discolouration (a sign of necrosis; due to which a maximum grade 4 was given for erythema).

There was no evidence of a corrosive effect on the skin.

 

On the skin exposed to the test substance for 1 and 4 hours, remnants of the test substance were present, being sticky on Day 1, and 24, 48 and 72 hours after exposure, and dry at 7 days after exposure. This did not hamper the scoring of the skin irritation reactions.

No evidence of full thickness destruction of the skin or scar tissue was observed during the observation period, indicating that no corrosion of the skin had occurred by dermal application of Amines, C36-alkylenedi- to the intact rabbit skin.

The extreme response following a 4-hour exposure to Amines, C36-alkylenedi- in a single animal was irreversible within the 14-day observation period.

 

Based on these results and according to the:

- Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2007), Amines, C36-alkylenedi- should be classified as : skin irritant (Category 2).

- Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures, Amines, C36-alkylenedi- should be classified as Irritant (Category 2) and labeled as H315: Causes skin irritation.