Amines, C36-alkylenedi-

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

29 March - 19 April 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 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye 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

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

- Source: Charles River France, L’Arbresle Cedex, France
- Age at study initiation: Animal used within the study was at least 6 weeks old
- Weight at study initiation: Body weights 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 Harlan Teklad, Italy). Hay and wooden sticks were available 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. Any variations to these conditions were maintained in the raw data and had no effect on the outcome of the study.

IN-LIFE DATES: From: 29 March - 19 April 2012

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: One eye of each animal remained untreated and served as the reference control.

Amount / concentration applied:

TEST MATERIAL- Amount(s) applied: 0.1 mL

Duration of treatment / exposure:

Single instillation on Day 1.

Observation period (in vivo):

The eyes of each animal were examined approximately 1, 24, 48 and 72 hours and 7, 14 and 21 days after instillation of the test substance

Number of animals or in vitro replicates:

1 male

Details on study design:

STUDY DESIGN
The study was performed in a stepwise manner and was started by treatment of a single rabbit (sentinel). Based on the nature and persistency of the ocular lesions observed during the study, the two further rabbits assigned to the study were not treated.

TREATMENT
On test Day 1, 0.1 mL of the test substance was instilled in the conjunctival sac of an eye of one animal after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second to prevent loss of the test substance.
Immediately after the 24-hour observation, a solution of 2% fluorescein (Merck, Darmstadt, Germany) in water (adjusted to pH 7.0) was instilled into both eyes of the animal to quantitatively determine corneal epithelial damage. This procedure was repeated to assess recovery. Any bright green stained area, indicating epithelial damage, was estimated as a percentage of the total corneal area. Immediately after fluorescein examination on Day 2, the treated eye was rinsed with approx 100 mL tepid tap water, using a velocity of flow which did not affect the eye, in an attempt to remove residual test substance. For reference control the other eye was also rinsed. After the final observation, the animal was sacrificed by intra-venous injection of Euthasol® 20% (AST Farma BV, Oudewater, The Netherlands).
The in vivo test with one rabbit showed that the (test) substance present at 24 hours after instillation could only partly be removed from the eye by rinsing after 24 hours. Based on the results obtained for this animal, an ex-vivo test was conducted to assess the effectiveness of rinsing at 1 hour postinstillation (also in relation to worker exposure risk assessment). The severity of the eye lesions observed in the treated animal did not allow this test to be conducted in vivo in first instance.

REMOVAL OF TEST SUBSTANCE
-Washing: Yes, at 24 hours after instillation with 100 ml tepid tap water

OBSERVATIONS
- Mortality/Viability: Twice daily.
- Toxicity: At least once daily.
- Body Weight: Day of treatment (prior to instillation) and after the final observation.
- Necropsy: No necropsy was performed according to protocol.
- Irritation: The eyes of each animal were examined approximately 1, 24, 48 and 72 hours and 7, 14 and 21 days after instillation of the test substance. The irritation scores and a description of all other (local) effects were recorded.The irritation was assessed according to OECD 405.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Remarks:

(opacity)

Basis:

mean

Time point:

other: 24, 48 and 72 hrs.

Score:

2

Max. score:

4

Results 2

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 24, 48 and 72 hrs.

Max. score:

2

Remarks on result:

other: Iris could not be scored due to (sticky) substance/secretions.

Results 3

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

mean

Time point:

other: 24, 48 and 72 hrs.

Score:

3

Max. score:

3

Results 4

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: 24, 48 and 72 hrs.

Score:

3

Max. score:

4

Irritant / corrosive response data:

Irritation: Instillation of 0.1 mL of Amines, C36-alkylenedi- into an eye of one rabbit resulted in effects on the cornea, iris and conjunctivae. The corneal injury consisted of opacity (maximum grade 3) and epithelial damage (maximum 65% of the corneal area). As a result of the corneal injury, pannus (neovascularization of the cornea) was apparent at 7, 14 and 21 days after instillation. Between 1 hour and 14 days after instillation, only approximately 25% of the cornea could be scored due to presence of (sticky) substance/secretions. The iris showed no reaction to a light stimulus at 1 hour after instillation. Iridial irritation/responses could not be scored at 24, 48 and 72 hours, and 7 and 14 days after instillation due to presence of (sticky) substance/secretions. The irritation of the conjunctivae consisted of redness, chemosis and discharge. Redness remained present up to 21 days after instillation. Reduced flexibility of the eyelids was noted at 72 hours, and 7, 14 and 21 days after instillation, and a bald skin of the eyelids was noted at 7 and 14 days after instillation.

Corrosion: There was no evidence of ocular corrosion.

Other effects:

Colouration/Remnants: (Sticky) substance/secretions were noted on the cornea at 1, 24, 48 and 72 hours, and 7 and 14 days after instillation. Remnants of the test substance were present on the outside of the eyelids at 1, 24, 48 and 72 hours after instillation. The eyes were rinsed at 24 hours following instillation. An ex vivo test showed that the remnants could also not be removed from the cornea at 1 hour following instillation.

Toxicity/Mortality: No symptoms of systemic toxicity were observed in the animal during the test period and no mortality occurred.

Applicant's summary and conclusion

Interpretation of results:

Category 1 (irreversible effects on the eye)

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Based on these results:
- according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2011), Amines, C36-alkylenedi- should be classified as : having irreversible effects on the eyes (Category 1).
- according to the Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures, Amines, C36-alkylenedi- should be classified as Irreversible effects on the eye (Category 1) and labeled as H318: Causes serious eye damage.

Executive summary:

Acute eye irritation/corrosion study with Amines, C36-alkylenedi- was performed in the rabbit, according to OECD 405 and under GLP.

A single sample of 0.1 mL of Amines, C36-alkylenedi- was instilled into an eye of one rabbit. Observations were made 1, 24, 48 and 72 hours and 7, 14 and 21 days after instillation. Based on the severity and persistency of the eye lesions, no further animals were treated.

Instillation of the test substance resulted in effects on the cornea, iris and conjunctivae.

The corneal injury consisted of opacity (maximum grade 3) and epithelial damage (maximum 65% of the corneal area). As a result of the corneal injury, pannus (neovascularization of the cornea) was apparent at 7, 14 and 21 days after instillation. Between 1 hour and 14 days after instillation, only approximately 25% of the cornea could be scored due to presence of (sticky) substance/secretions.

The iris showed no reaction to a light stimulus at 1 hour after instillation. Iridial irritation/responses could not be scored at 24, 48 and 72 hours, and 7 and 14 days after instillation due to presence of (sticky) substance/secretions. The irritation of the conjunctivae consisted of redness, chemosis and discharge. Redness remained present up to 21 days after instillation. Reduced flexibility of the eyelids was noted at 72 hours, and 7, 14 and 21 days after instillation, and a bald skin of the eyelids was noted at 7 and 14 days after instillation.

There was no evidence of ocular corrosion.

(Sticky) substance/secretions were noted on the cornea at 1, 24, 48 and 72 hours, and 7 and 14 days after instillation. Remnants of the test substance were present on the outside of the eyelids at 1, 24, 48 and 72 hours after instillation. The eyes were rinsed at 24 hours following instillation.

Anex vivotest showed that the remnants could also not be removed from the cornea at 1 hour following instillation.

 

According to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2011), Amines, C36-alkylenedi- should be classified as : having irreversible effects on the eyes (Category 1).