L-Alanine, N-coco acyl derivs., sodium salts

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2006

Report date:

2006

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

2002

Deviations:

no

GLP compliance:

not specified

Test material

Test material information

Constituent 1

Reference substance name:

L-Alanine, N-coco acyl derivs., sodium salts

EC Number:

290-478-9

EC Name:

L-Alanine, N-coco acyl derivs., sodium salts

Cas Number:

90170-45-9

Molecular formula:

RHNNaO3

IUPAC Name:

L-Alanine, N-coco acyl derivs., sodium salts

Test material form:

solid

Specific details on test material used for the study:

Test item 1
Name: Sodium N-cocoyl-L-alaninate (ACS-12)
Lot No.: 051020-2
Purity: 30.2% (residue: water 69.8%)
Description: liquid (pH 8.4 / 20°C)
Expiration: October 30, 2006

Test item 2
Name: Sodium N-cocoyl-L-alaninate (ACS-12)
Lot No.: 601011
Purity: 30.1% (residue: water 69.9%)
Description: liquid (pH 8.7 / 20°C)
Expiration: January 31, 2007

Test animals / tissue source

Species:

rabbit

Strain:

other: Japanese White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Japan Laboratory Animals Inc.
- Age at study initiation: 13 weeks
- Weight at study initiation: 2.49 – 2.78 kg
- Housing: individually
- Diet: ad libitum
- Water: ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 – 24
- Humidity (%): 44 – 79
- Photoperiod (hrs dark / hrs light): 12 / 12

Test system

Vehicle:

water

Controls:

no

Amount / concentration applied:

0.1 mL

Duration of treatment / exposure:

single application

Observation period (in vivo):

up to 6 days

Number of animals or in vitro replicates:

3 f per lot

Details on study design:

The rabbits’ eyes were examined with an ophthalmoscope on the day before application of the test articles. Six animals with no ocular abnormalities, showing regular body weight gain during the quarantine/acclimation period and showing no clinical abnormalities, were chosen and used for the study. At the time of eye examination, the cornea was also examined, using 2% fluorescein sodium aqueous solution, for the presence or absence of abnormalities.
The test items were applied in the conjunctival sac of the left eye of each animal after gently pulling the lower eyelid away from the eyeball. The lid was then gently held together for about 1 second in order to prevent loss of the test article. The right eye, which remained untreated, served as a control.
Animals were observed for clinical signs immediately after application and hourly up to 6 hours after application and once daily thereafter up to day 6.
All animals were weighed on the day of application and on the final day of observation.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

maximum mean total score (MMTS)

Remarks:

results for test item 1

Basis:

mean

Time point:

24 h

Score:

18.7

Max. score:

110

Reversibility:

fully reversible within: 5 days

Remarks on result:

probability of moderate irritation

Results 2

Irritation parameter:

maximum mean total score (MMTS)

Remarks:

results for test item 2

Basis:

mean

Time point:

24 h

Score:

23.3

Max. score:

110

Reversibility:

fully reversible within: 6 days

Remarks on result:

probability of moderate irritation

Irritant / corrosive response data:

Test item 1
Corneal opacity, conjunctival redness, chemosis and discharge were observed in all (3/3) animals from 1 hour after application. At 24 hours after application, chemosis and discharge diminished but the area of corneal opacity and conjunctival redness increased in 2/3 or all animals and 1/3 animals developed iridal changes. After that, eye irritation reactions were reduced gradually, and iridal changes and chemosis disappeared by 48 hours after application, discharge disappeared by 72 hours after application and corneal opacity disappeared by 96 hours after application. Conjunctival redness was observed only in 1/3 animals at 96 hours after application, but it disappeared by day 5 after application. As other ocular changes, eyelid closure was observed from immediately to 6 hours after application in all animals. The MMTS (Maximum mean total score) was 18.7 at 24 hours after application.

Test item 2
Corneal opacity, conjunctival redness, chemosis and discharge were observed in all (3/3) animals from 1 hour after application. At 24 hours after application, chemosis diminished but the area of corneal opacity and conjunctival redness increased in all animals. After that, eye irritation reactions were reduced gradually, and chemosis disappeared by 48 hours after application and discharge disappeared by 72 hours after application. Conjunctival redness was observed in all animals and corneal opacity was observed in 1/3 animals at 96 hours after application, but they disappeared by day 6 after application. As other ocular changes, eyelid closure was observed in all animals from immediately to 6 hours after application and in 2/3 animals on day 1 after application. The MMTS was 23.3 at 24 hours after application.

Applicant's summary and conclusion

Interpretation of results:

Category 2B (mildly irritating to eyes) based on GHS criteria

Conclusions:

An eye irritation study was conducted in female Japanese White rabbits, in order to examine the irritant effects of two lots of ACS-12 on the eye. 0.1 mL of the diluted test items (5 % w/w) was applied to the left eye of three rabbits for each lot. The cornea, iris and conjunctiva were observed periodically, and irritation reactions were evaluated according to the Draize criteria. Eye irritation reactions were categorized and determined on the basis of the Kay and Calandra method.
Based on the results of this study it was concluded that the two test items should be classified as moderately irritating on the rabbit eye.

Executive summary:

An eye irritation study was conducted in female Japanese White rabbits, in order to examine the irritant effects of two lots of ACS-12 on the eye. 0.1 mL of the diluted test items (5 % w/w) was applied to the left eye of three rabbits for each lot. The cornea, iris and conjunctiva were observed periodically, and irritation reactions were evaluated according to the Draize criteria. Eye irritation reactions were categorized and determined on the basis of the Kay and Calandra method.

Based on the results of this study it was concluded that the two test items should be classified as moderately irritating on the rabbit eye.