Reaction mass of Amines, coco alkyl and β-Alanine, N-(2-carboxyethyl)-, N-coco alkyl derivs. and β-Alanine, N-coco alkyl derivs.

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

12 June 2012 - 17 July 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Compliant to GLP and testing guidelines; adequate consistence between data, comments and conclusions.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

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

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of Amines, coco alkyl and ß-Alanine, N-(2-carboxyethyl)-, N-coco alkyl derivs. and ß-Alanine, N-coco alkyl derivs.

EC Number:

915-790-0

Molecular formula:

n/a

IUPAC Name:

Reaction mass of Amines, coco alkyl and ß-Alanine, N-(2-carboxyethyl)-, N-coco alkyl derivs. and ß-Alanine, N-coco alkyl derivs.

Test material form:

other: paste

Details on test material:

- Lot/batch No.: EL 0085
- Name of test material: Reaction mass of Amines, coco alkyl and ß-Alanine, N-(2-carboxyethyl)-, N coco alkyl derives. and ß-Alanine, N coco alkyl derivs.
- Physical state: solid paste
- Expiry date: 15 May 2015
- Storage condition: in a cool place.

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: breeder: CEGAV, Argenvilliers, France
- Age at study initiation: 2 to 4 months old on the day of treatment
- Mean body weight at study initiation: 3070 g or 3145 g
- Fasting period before study: no
- Housing: one animal was individually housed in Pajon cages (57 cm x 75 cm x 50 cm) and the other animal was individually housed in noryl cages (Tecniplast, 65.3 cm x 65.3 cm x 45 cm)
- Diet: 110 pelleted diet (free access)
- Water: tap water filtered with a 0.22 µm filter (free access)
- Acclimation period: at least 5 days before the beginning of the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 ± 3°C
- Humidity (%): 50 ± 20%
- Air changes (per hr): approximately 12 cycles/hour of filtered, non-recycled air
- Photoperiod (hrs dark / hrs light): 12 h/12 h.

IN-LIFE DATES: 20 June 2012 to 17 July 2012.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: untreated right eye served as a control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 0.1 g/animal.

Duration of treatment / exposure:

Not applicable: single application not followed by rinsing.

Observation period (in vivo):

1, 24, 48 and 72 h; if relevant, daily until reversibility of reactions

Number of animals or in vitro replicates:

Two males.

Details on study design:

REMOVAL OF TEST SUBSTANCE: No

SCORING SYSTEM: Draize scale.

- Conjunctival chemosis (lids and/or nictitating membranes):
0 no swelling
1 any swelling above normal (includes nictitating membranes)
2 obvious swelling with partial eversion of lids
3 swelling with lids about half-closed
4 swelling with lids more than half-closed

- Conjunctival redness (palpebral and bulbar conjunctivae, cornea and iris):
0 blood vessels normal
1 a number of blood vessels definitely hyperemic (injected)
2 diffuse, crimson colour, individual vessels not easily discernible
3 diffuse, beefy red

- Discharge:
0 absence of discharge
1 slight discharge (does not include small amounts normally found in inner canthus)
2 discharge with moistening of lids and hairs adjacent to lids
3 discharge with moistening of lids and hairs on wide area around the eye

- Iris lesions
0 normal
1 markedly deepened rugae, congestion, swelling, moderate circum-corneal hyperemia,or injection, any of these or combination of any thereof, iris still reacting to light (sluggish reaction is positive)
2 no reaction to light, haemorrhage, gross destruction (any or all of these)

- Cornea intensity of opacity (direct examination and, if necessary, with an UV lamp)
0 no ulceration or opacity
1 scattered or diffuse areas of opacity (other than slight dulling or normal lustre), details of iris clearly visible
2 easily discernible translucent area, details of iris slightly obscured
3 nacreous areas, no details of iris visible, size of pupil barely discernible
4 opaque cornea, iris not discernible through the opacity

- Cornea area of opacity (direct examination and, if necessary, with an UV lamp)
1 one quarter (or less) but not zero
2 greater than one quarter but less than a half
3 greater than one half but less than three quarters
4 greater than three quarters up to whole area

- Any other lesions observed were noted

TOOL USED TO ASSESS SCORE: UV lamp after instillation of 0.5% sodium fluorescein solution

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Remarks:

(intensity)

Basis:

animal #1

Time point:

other: 24, 48 and 72 h (mean)

Score:

2

Max. score:

2

Reversibility:

fully reversible within: Day 13

Results 2

Irritation parameter:

cornea opacity score

Remarks:

(intensity)

Basis:

animal #2

Time point:

other: 24, 48 and 72 h (mean)

Score:

3

Max. score:

3

Reversibility:

fully reversible within: Day 19

Results 3

Irritation parameter:

iris score

Basis:

animal #1

Time point:

other: 24, 48 and 72 h (mean)

Score:

1

Max. score:

1

Reversibility:

fully reversible within: Day 11

Results 4

Irritation parameter:

iris score

Basis:

animal #2

Time point:

other: 24, 48 and 72 h (mean)

Score:

0.3

Max. score:

1

Reversibility:

fully reversible within: Day 3

Results 5

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #1

Time point:

other: 24, 48 and 72 h (mean)

Score:

3

Max. score:

3

Reversibility:

fully reversible within: Day 14

Results 6

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #2

Time point:

other: 24, 48 and 72 h (mean)

Score:

3

Max. score:

3

Reversibility:

fully reversible within: Day 22

Results 7

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: 24, 48 and 72 h (mean)

Score:

3

Max. score:

3

Reversibility:

fully reversible within: Day 16

Results 8

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

other: 24, 48 and 72 h (mean)

Score:

2.3

Max. score:

3

Reversibility:

fully reversible within: Day 22

Applicant's summary and conclusion

Interpretation of results:

irritating

Remarks:

Migrated information severely irritant Criteria used for interpretation of results: other: CLP Regulation

Conclusions:

The test item was severely irritant when administered by ocular route to rabbits.

According to the criteria of CLP Regulation, the test item should be classified category 1 and assigned the signal word “danger” and the hazard statement “H318: causes serious eye damage”.

Executive summary:

The objective of this study was to evaluate the potential irritant properties of the test item for the eye following a single administration to rabbits.

This study was conducted in compliance with the principles of Good Laboratory Practice.

Methods

The test item was first administered to a single male New Zealand White rabbit.

As mean value from grading at 24, 48 and 72 hours after instillation was = 2 for conjunctival edema (chemosis) or for conjunctival redness and/or = 1 for iris lesions or for corneal opacity, the test item was administered in the left eye of a second animal.

After administration to the second animal, asmean value from grading at 24, 48 and 72 hours after instillation was = 2 for conjunctival edema (chemosis) or for conjunctival redness and/or = 1 for iris lesions or for corneal opacity, no other animal was treated.

The test item was administered inthe conjunctival sac of the left eye. The right eye remained untreated and served as control.

A quantity of 0.1 g/animal was used.

The eyes were not rinsed just after the 1-hour scoring.

Each animal was observed at least once a day for mortality and clinical signs. Ocular reactions were observed approximately 1 hour, 24, 48 and 72 hours after the administration and then daily until the end of the observation period. The mean values of the scores for chemosis, redness of the conjunctiva, iris lesions and corneal opacity were calculated for each animal. Body weight was recorded on the day of treatment and the end of the observation period.

On completion of the observation period, the animals were sacrificed then discarded without macroscopic post-mortem examination.

Results

No unscheduled deaths occurred during the study and no clinical signs were noted in any animals.

The body weight of the animals was unaffected by the test item treatment.

No ocular reactions were observed in the right untreated control eye.

 

In the left treated eye of one animal, marked chemosis and severe redness of the conjunctiva were observed from day 1 to day 4 and then, slight or moderate reactions persisted until day 15. Iris lesions were noted from day 1 to day 4 and then on day 10. Moderate corneal opacity was recorded from day 2 to day 10 and then, slight reactions persisted up to day 12.

In addition, lacrimation was noted from day 2 to day 5, whitish thick discharge on days 2 and 10 and neovascularisation from day 9 to day 11.

In the left eye of the other animal, moderate or marked chemosis and severe redness of the conjunctiva were observed from day 1 to day 6 and then slight or moderate reactions persisted up to day 21. Iris lesions were noted on day 2 only. Marked corneal opacity, which covered more than three quarters up to the whole corneal area on days 2 and 3, was recorded from day 2 to day 6 and then, moderate reactions were observed up to day 17 and slight corneal opacity persisted on day 18.

In addition, lacrimation and whitish thick discharge were noted between days 1 and 15 and neovascularisation persisted from day 7 until the end of the observation period.

Mean scores calculated for each animal over 24, 48 and 72 hours were as follows:

.       chemosis : 3.0 and 2.3; showing eye irritation,

.       redness of the conjunctiva: 3.0 and 3.0; showing eye irritation,

.       iris lesions: 1.0 and 0.3; showing eye irritation in 1/2 animals,

.       corneal opacity: 2.0 and 3.0; showing eye irritation.

 

As neovascularization was observed until the end of the observation period (day 22) in one animal, it was considered that the test item caused serious eye irritation.

 

Conclusion

The test item was severely irritant when administered by ocular route to rabbits.

 

According to the criteria of CLP Regulation,the test item should be classified category 1 and assigned the signal word “danger” and the hazard statement “H318: causes serious eye damage”.