LUMIPOD

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

16/03/2020

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2020

Report date:

2020

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Certificat n°2019/19

Test material

Test material information

Constituent 1

Reference substance name:

L-Leucine, reaction products with 1,4:3,6-dianhydro-D-glucitol, iso-Pr alc. and octanoyl chloride

Molecular formula:

Not applicable

IUPAC Name:

L-Leucine, reaction products with 1,4:3,6-dianhydro-D-glucitol, iso-Pr alc. and octanoyl chloride

Test animals / tissue source

Species:

other: bovine cornea

Strain:

other: not applicable

Details on test animals or tissues and environmental conditions:

Corneas of calves aged less than 8 months. Calves eyes were collected in the slaughterhouse of Sobeval Boulazac 24759 - France, in a short time after slaughtering of the animals, and transported in a Hanks's buffered saline solution with antibiotic to the laboratory At reception, the eyes were carefully examined under lighting and these showing a visible (scratches, pigmentation, neo-vascularization) defect were eliminated.
For each selected eye, an incision with a scalpel was practiced at the level of the scleral ring by means of scissors.
Approximately 2 to 3 mm of scleral ring were left to facilitate the further handlings.
Corneas were immersed in Hanks's medium at room temperature.
Then the corneas were used at receipt for the test.
The demonstration of the skill of the laboratory is performed using the test substances recommended by the guideline followed.

Test system

Vehicle:

not specified

Controls:

yes

Amount / concentration applied:

750 +- 75 mg of the test substance

Duration of treatment / exposure:

The treatment lasted (time of exposure) was 10 minutes.
After the end of the exposure period, precisely signalled by the ringing of the chronometer, the test item and controls are removed from the anterior compartment and the epithelium washed at least three times (or until no visual evidence of test item can be observed) with EMEM containing phenol red.

Duration of post- treatment incubation (in vitro):

The opacity of each cornea was measured at 2 times, just before treatment with the test item (measurement called OPTO) and 2 hours after the end of the 10 minutes contact (measurement called OPT2).

Number of animals or in vitro replicates:

Not applicable.
Triplicate corneas for each timepoint and tested substance (test item, vehicle control, positive control).

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Run / experiment:

Mean of three runs with two replicates each

Value:

>= 15.1 - <= 16.5

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Applicant's summary and conclusion

Conclusions:

According to the defined grading scale, no prediction can be made for the classification of the test item L-Leucine- reaction products with 1-4:3-6-dianhydro-D-glucitol- iso-Pr alc. and octanoyl chloride - Batch : 200122016662 concerning eye irritation or serious eye damage.

Executive summary:

PRINCIPLE OF THE STUDY
The aim of the study was to assess quantitatively the irritant potential of a test item after application to the isolated calf cornea.
The assessment was based on the measurement of two parameters: the opacity and permeability of the cornea whose deteriorations reflected the damage of the tissue.
The test item was let in contact with the isolated cornea for 10 minutes.
On the basis of the results obtained, an In Vitro Irritancy score (IVIS) was calculated to classify the irritancy level of the test item.

EXPERIMENTAL STARTING DATE AND EXPERIMENTAL COMPLETION DATE: March 16, 2020

RESULTS:

Test item	Tested concentration	IVIS 10 minutes Mean ± standard deviation
L-Leucine- reaction products with 1- 4:3-6-dianhydro-D-glucitol- iso-Pr alc. and octanoyl chloride Batch : 200122016662	as supplied	15.8 ± 0.7

No prediction can be made for the classification of the test item L-Leucine- reaction products with 1-4:3-6-dianhydro-D-glucitol- iso-Pr alc. and octanoyl chloride - Batch ; 200122016662 concerning eye irritation or serious eye damage.