Registration Dossier

Toxicological information

Eye irritation

Currently viewing:

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
28 April 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report Date:
2016

Materials and methods

Test guideline
Qualifier:
according to
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
Principles of method if other than guideline:
There were no deviations from standard operating procedures that affected the integrity of the study.
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Specific details on test material used for the study:
Name (as stated in the report): Freskomenthe
Batch: VE00411894
Expiration date: February 11, 2018

Test animals / tissue source

Species:
other: Bovine
Details on test animals or tissues and environmental conditions:
Test System: Bovine eyes were used as soon as possible after slaughter.
Rationale: In the interest of sound science and animal welfare, a sequential testing strategy is recommended to minimise the need of in vivo testing (1-6). As a consequence a validated and accepted in vitro test for eye irritation should be performed before in vivo tests are conducted. One of the proposed validated in vitro eye irritation tests is the Bovine Corneal Opacity and Permeability (BCOP) test.
Source: Bovine eyes from young cattle were obtained from the slaughterhouse (Vitelco, 's Hertogenbosch, The Netherlands), where the eyes were excised by a slaughterhouse employee as soon as possible after slaughter.
Transport: Eyes were collected and transported in physiological saline in a suitable container under cooled conditions.

Test system

Vehicle:
unchanged (no vehicle)
Remarks:
The test item was tested neat.
Controls:
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
Undiluted FRESKOMENTHE
Duration of treatment / exposure:
The test item was tested undiluted
Details on study design:
The medium from the anterior compartment was removed and 750 μl of either the negative control, positive control (Ethanol) or test item was introduced onto the epithelium of the cornea. The holders were slightly rotated, with the corneas maintained in a horizontal position, to ensure uniform distribution of the control or the test item over the entire cornea. Corneas were incubated in a horizontal position for 10 ± 1 minutes at 32 ± 1°C. After the incubation the solutions were removed and the epithelium was washed with MEM with phenol red (Earle’s Minimum Essential Medium, Life Technologies) and thereafter with cMEM. Possible pH effects of the test item on the corneas were recorded. The medium in the posterior compartment was removed and both compartments were refilled with fresh cMEM. Subsequently the corneas were incubated for 120 ± 10 minutes at 32 ± 1°C. After the completion of the incubation period opacity determination was performed. Each cornea was inspected visually for dissimilar opacity patterns.

Results and discussion

In vitro

Results
Irritation parameter:
cornea opacity score
Remarks:
mean in vitro irritancy score after 10 minutes of treatment
Value:
8.4
Negative controls validity:
valid
Remarks:
0.9
Positive controls validity:
valid
Remarks:
46.2

Any other information on results incl. tables

The individual in vitro irritancy scores for the negative controls ranged from 0.7 to 1.1. The individual positive control in vitro irritancy scores ranged from 41 to 55 for Ethanol. The corneas treated with the positive control item were turbid after the 10 minutes of treatment.

The corneas treated with FRESKOMENTHE showed opacity values ranging from 5.9 to 9.2 and permeability values ranging from 0.028 to 0.173. The corneas were clear after the 10 minutes of treatment with FRESKOMENTHE. No pH effect of the test item was observed on the rinsing medium. Hence, the in vitro irritancy scores ranged from 6.7 to 9.9 after 10 minutes of treatment with FRESKOMENTHE.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The negative control responses for opacity and permeability were less than the upper limits of the laboratory historical range indicating that the negative control did not induce irritancy on the corneas. The mean in vitro irritancy score of the positive control (Ethanol) was 46 and was within two standard deviations of the current historical positive control mean. It was therefore concluded that the test conditions were adequate and that the test system functioned properly.
FRESKOMENTHE induced ocular irritation through both endpoints, resulting in a mean in vitro irritancy score of 8.4 after 10 minutes of treatment.
Since FRESKOMENTHE induced an IVIS > 3 ≤ 55, no prediction on the classification can be made.
Executive summary:

Evaluation of the eye hazard potential of FRESKOMENTHE using the Bovine Corneal Opacity and Permeability test (BCOP test).

This report describes the potency of chemicals to induce serious eye damage using isolated bovine corneas. The eye damage of FRESKOMENTHE was tested through topical application for 10 minutes.

The study procedures described in this report were based on the most recent OECD guideline.

Batch VE00411894 of FRESKOMENTHE was a colourless liquid with a purity of 99.1%. The test item was applied as it is (750 μl) directly on top of the corneas.

The negative control responses for opacity and permeability were less than the upper limits of the laboratory historical range indicating that the negative control did not induce irritancy on the corneas. The mean in vitro irritancy score of the positive control (Ethanol) was 46 and was within two standard deviations of the current historical positive control mean. It was therefore concluded that the test conditions were adequate and that the test system functioned properly.

FRESKOMENTHE induced ocular irritation through both endpoints, resulting in a mean in vitro irritancy score of 8.4 after 10 minutes of treatment.

Since FRESKOMENTHE induced an IVIS > 3 ≤ 55, no prediction on the classification can be made.