Disodium 2,4-diamino-3-{[4-(ethenylsulfonyl)phenyl]diazenyl}-5-{[4-(ethenylsulfonyl)-2-sulfonatophenyl]diazenyl}benzenesulfonate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)

Deviations:

no

Principles of method if other than guideline:

Guidelines followed

GLP compliance:

yes

Test material

Test material information

Constituent 1

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

None

Test animals / tissue source

Species:

other: cattle

Strain:

not specified

Details on test animals or tissues and environmental conditions:

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:

water

Controls:

not specified

Amount / concentration applied:

No correction was made for the purity/composition of the test substance.

A 20% (w/w) solution of FAT 40866/A TE was prepared in physiological saline (Merck, Darmstadt, Germany). The stock solution was treated with ultrasonic waves until the test substance had completely dissolved.

Duration of treatment / exposure:

The medium from the anterior compartment was removed and 750 µl of either the negative control, positive control (20% (w/v) Imidazole solution) or 20% (w/w) test substance was introduced onto the epithelium of the cornea. The holder was slightly rotated, with the corneas maintained in a horizontal position, to ensure uniform distribution of the solutions over the entire cornea. Corneas were incubated in a horizontal position for 240 ± 10 minutes at 32 ± 1ºC. After the incubation the solutions were removed and the epithelium was washed at least three times with MEM with phenol red (Eagle’s Minimum Essential Medium, Invitrogen Corporation). Possible pH effects of the test substance on the corneas were recorded. The anterior and the posterior compartment were refilled with fresh cMEM and an opacity determination was performed without any further incubation. After the completion of the incubation period each cornea were inspected visually for dissimilar opacity patterns and the opacity determination was performed. To verify the results a repeat experiment was performed (Experiment 2).

Observation period (in vivo):

None

Number of animals or in vitro replicates:

None

Details on study design:

Acceptability of the assay:

The assay is considered acceptable if:
- The positive control gives an in vitro irritancy score that falls within the laboratory historical mean value.
- The negative control responses should result in opacity and permeability values that are less than the upper limits of the laboratory historical range.

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

in vitro irritation score

Run / experiment:

Mean In vitro Irritation Score

Value:

3.6

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

cornea opacity score

Run / experiment:

Mean In vitro Irritation Score

Value:

0.6

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

In vivo

Irritant / corrosive response data:

None

Other effects:

None

Any other information on results incl. tables

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 (20% (w/v) Imidazole) was 115 in experiment 1 and 128 in experiment 2 and within the historical positive control data range.

It was therefore concluded that the test conditions were adequate and that the test system functioned properly. FAT 40866/A TE showed in vitro irritancy scores of 0.6, 0.5 and 10.8 after 240 minutes of treatment. Since one out of three corneas showed an IVIS above 3, the test was repeated to confirm the results, resulting in IVIS scores of -0.2 to 1.0. Therefore the value of 10.8 observed in the first experiment can be considered an outlier.

Finally, it is concluded that this test is valid and that FAT 40866/A TE is not irritant or corrosive in the Bovine Corneal Opacity and Permeability test under the experimental conditions described in this report.

Since FAT 40866/A TE induced an IVIS ≤ 3, no classification is required for eye irritation or serious eye damage.

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: OECD GHS

Conclusions:

It is concluded that this test is valid and that FAT 40866/A TE is not irritant or corrosive in the Bovine Corneal Opacity and Permeability test under the experimental conditions described in this report.

Executive summary:

Screening for the eye irritancy potential of FAT 40866/A TE using the Bovine Corneal Opacity and Permeability test (BCOP test).

This report describes the ocular irritation properties of FAT 40866/A TE on an isolated bovine cornea.

The possible ocular irritancy of FAT 40866/A TE was tested through topical application for approximately 240 minutes.

The study procedures described in this report were based on the most recent OECD and EC guidelines.

Batch BOP 04-13 (Lot: BS-MIF 975) of FAT 40866/A TE was a red-brown powder. The test substance was applied as a 20% (w/w) solution (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 (20% (w/v) Imidazole) was 115 (experiment 1) and 128 (experiment 2) and within the historical positive control data range in two independent experiments. It was therefore concluded that the test conditions were adequate and that the test system functioned properly.

FAT 40866/A TE showed in vitro irritancy scores of 0.6, 0.5 and 10.8 after 240 minutes of treatment. Since one out of three corneas showed an IVIS above 3, the test was repeated to confirm the results, resulting in IVIS scores of -0.2 to 1.0. Therefore the value of 10.8 observed in the first experiment can be considered an outlier.

Finally, it is concluded that this test is valid and that FAT 40866/A TE is not irritant or corrosive in the Bovine Corneal Opacity and Permeability test under the experimental conditions described in this report.

Since FAT 40866/A TE induced an IVIS ≤ 3, no classification is required for eye irritation or serious eye damage.