C16-18-(even numbered)-alkyl fatty acid, compound with C16-18-(even numbered)-alkylamine

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

07 March 2013 -- 03 April 2013

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:

2013

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

yes

Remarks:

Irritation parameters were not evaluated on day 22 for the first animal. As the more severe classification has been applied, this deviation was not considered to have compromised the validity of the study.

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

see above

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

C16-18-(even numbered)-alkyl fatty acid, compound with C16-18-(even numbered)-alkylamine

EC Number:

800-984-9

Cas Number:

1428547-35-6

Molecular formula:

RNH3+, R’COO- where R = C16-18-alkyl and R' = C15-17-alkyl

IUPAC Name:

C16-18-(even numbered)-alkyl fatty acid, compound with C16-18-(even numbered)-alkylamine

Test material form:

other: light yellow solid

Details on test material:

- Other name: N-(hydrogenated tallow alkyl) amine, N-(hydrogenated tallow alkyl) fatty acid salt without free fatty acid
- Physical state: light yellow solid
- Lot/batch No.: EP 4756
- Analytical purity: 100%
- Expiry date: 10 January 2015
- Storage condition: at room temperature

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: 2410 g and 2775 g
- Fasting period before study: no
- Housing: individually housed in noryl cages
- Diet: 110 pelleted diet (free access)
- Water: tap water filtered with a 0.22 µm filter (free access)
- Acclimation period: a period of 4 or 6 days before treatment .

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: 18 March 2013 to 03 April 2013

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: 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:

chemosis score

Basis:

animal #1

Time point:

other: 24, 48 and 72 h (mean)

Score:

3.3

Max. score:

4

Reversibility:

not fully reversible within: day 21

Remarks on result:

other: Day 22: irritation parameters not evaluated

Results 2

Irritation parameter:

other: conjunctivae score (redness)

Basis:

animal #1

Time point:

other: 24, 48 and 72 h (mean)

Score:

3

Max. score:

3

Reversibility:

not fully reversible within: day 21

Remarks on result:

other: Day 22: irritation parameters not evaluated

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 20

Remarks on result:

other: Day 22: irritation parameters not evaluated

Results 4

Irritation parameter:

cornea opacity score

Remarks:

(intensity)

Basis:

animal #1

Time point:

other: 24, 48 and 72 h (mean)

Score:

2

Max. score:

3

Reversibility:

fully reversible within: day 19

Remarks on result:

other: Day 22: irritation parameters not evaluated

Results 5

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

other: 24, 48 and 72 h (mean)

Score:

1.7

Max. score:

2

Reversibility:

fully reversible within: day 10

Results 6

Irritation parameter:

other: conjunctivae score (redness)

Basis:

animal #2

Time point:

other: 24, 48 and 72 h (mean)

Score:

2.7

Max. score:

3

Reversibility:

fully reversible within: day 10

Results 7

Irritation parameter:

iris score

Basis:

animal #2

Time point:

other: 24, 48 and 72 h (mean)

Score:

0.7

Max. score:

1

Reversibility:

fully reversible within: day 4

Results 8

Irritation parameter:

other: cornea score intensity

Basis:

animal #2

Time point:

other: 24, 48 and 72 h (mean)

Score:

2.7

Max. score:

3

Reversibility:

fully reversible within: day 8

Applicant's summary and conclusion

Interpretation of results:

other: severely irritant

Remarks:

Criteria used for interpretation of results: EU

Conclusions:

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

In addition, the effects on the conjunctiva were not fully reversible within 21 days of application in one animal; it was considered that the test item caused serious eye damage.

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 OECD Guideline No. 405 and the principles of Good Laboratory Practices.

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) and for conjunctival redness and = 1 for iris lesions and for corneal opacity, the test item was administered in the left eye of a second animal.

 

After administration to the second animal, as mean value from grading at 24, 48 and 72 hours after instillation was = 2 for conjunctival redness and = 1 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.

For the first and second animals, a local anesthetic was used before the 24-hour reading and prior to treatment, respectively.

 

Just after the 1-hour scoring, both eyes were rinsed with a sterile isotonic saline solution (0.9% NaCl).

Due to important ocular reactions in both animals and in order to perform ocular reactions evaluation in good conditions, eyes were rinsed again before the 24-hour reading with sterile isotonic saline solution (0.9% NaCl).

 

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 reversibility of the ocular reactions or until day 21. 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 at the end of the evaluation of ocular reactions or the day after the end of the evaluation of ocular reaction.

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.

One animal showed loud breathing on day 2. This could be considered as pain resulting from severity of ocular lesions. Thereafter chromorhynorrhea probably due to repeated administrations offluorescein solutionswas noted from day 15.

However no clinical signs indicative of systemic toxicity were observed in any animals. The body weight of the animals was unaffected by the test item treatment.

In the left treated eye of one animal, marked to severe chemosis and severe redness of the conjonctiva were observed from day 1 to day 4. Severe redness persisted on day 5. Slight to moderate conjunctiva reactions were noted between days 6 and 11. Thereafter, moderate to marked chemosis was recorded between days 12 and 19, whereas moderate to severe redness was observed from day 12 to day 16. Then, slight conjunctiva reactions persisted until the end of the observation period (day 21).

Iris lesions were noted from day 1 to day 19.

Moderate to marked corneal opacity was observed on days 2 and 3. Then, slight corneal opacity was recorded between days 4 and 14. Corneal opacity was graded marked again from day 15 to day 18.

In addition, marked blepharospasm was noted on day 2 and neovascularization was recorded from day 8 to day 18.

 

In the left treated eye of one animal, moderate chemosis and moderate to severe redness of the conjonctiva were noted from day 1 to day 3. Then, slight to moderate conjonctiva reactions persisted until day 9.

Iris lesions were noted from day 1 to day 3.

Moderate to marked corneal opacity was recorded from day 2 to day 4. Thereafter, slight corneal opacity was still present until day 7.

 

Lacrimation and whitish deposit were noted in the left eye of both animals on many occasions during the observation period.

 

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

.       chemosis: 3.3 and 1.7 ; showing eye irritation in 1/2 animals,

.       redness of the conjunctiva: 3.0 and 2.7; showing eye irritation in 2/2 animals,

.       iris lesions: 1.0 and 0.7 ; showing eye irritation in 1/2 animals,

.       corneal opacity: 2.0 and 2.7; showing eye irritation in 2/2 animals.

 

However, as the effects on the conjunctiva were not fully reversible within 21 days of application in one animal; it was considered that the test item caused serious eye damage.

 

Conclusion

 

Under the experimental conditions of this study, the test item was severely irritant when administered by ocular route to rabbits.

In addition, the effects on the conjunctiva were not fully reversible within 21 days of application in one animal; it was considered that the test item caused serious eye damage.

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".