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Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2012-11-05
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
The reliability is rated 1 because the study followed the standard guideline of reference (OECD 438), which describes a procedure designed to evaluate this endpoint. The results were reviewed for reliability and assessed as valid, and the study was conducted under GLP condition.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report date:
2013

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
EU method B.48 (Isolated chicken eye test method for identifying occular corrosives and severe irritants)
Qualifier:
according to guideline
Guideline:
OECD Guideline 438 (Isolated Chicken Eye Test Method for Identifying Ocular Corrosives and Severe Irritants)
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
anilino(phenylimino)methanaminium 3-[(4-anilinophenyl)diazenyl]benzenesulfonate
EC Number:
941-151-0
Molecular formula:
C31H28N6SO3
IUPAC Name:
anilino(phenylimino)methanaminium 3-[(4-anilinophenyl)diazenyl]benzenesulfonate
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Name of test material : Sepisol Fast Yellow MG-DPG
- Substance type: Organic salt
- Stability under test conditions: Stable
- Storage condition of test material: room temperature

Test animals / tissue source

Species:
other: eyes from chickens freshly killed
Details on test animals or tissues and environmental conditions:
TEST EYES
- Source: Slaughterhouses Etablissement Brun, 33820 Etaulier, France
- Age of the donors animals: approximately 7 weeks old
- Weight of the donors animals: between 1.5 to 2.5 kg
- Transport of eyes: The intact heads were transported from the slaughterhouse at ambient temperature in polystrene boxes humidified with towels moistened with isotonic saline. The eyes are enucleated in the laboratory within less than 1h30 after the the heads are removed
- Time between collection and use of corneas:The corneas were transported to the laboratory straight after removal on animals and used in a maximum delay of 24 hours after reception at the laboratory.

Test system

Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied : 30 mg
Duration of treatment / exposure:
Treatment of10 secondes
Observation period (in vivo):
4 hours post-treatment
Number of animals or in vitro replicates:
3 chicken eyes for the treatment group
3 chicken eyes for the positive control group
1 chicken eye for the negative control group
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes, with 40 mL of physiological saline (4 times for the treatement group and 2 times for the control groups)
- Time after start of exposure: after 10 secondes

SCORING SYSTEM: Determination of ICE classes for 3 endpoints (1- corneal thickness, 2- corneal opacitiy, 3- fluorescein retention) according to a predefine range. Each ICE classes are then combined to generate an Irritancy Classification

TOOL USED TO ASSESS SCORE:
- Corneal thickness : quantitative determination by optical pachymeter on a slit-lamp microscope.
- Corneal opacity: qualitative assessment based on the area of the cornea opacified
- Fluorescein retention : qualitative assessment based on the staining of the cornea

Results and discussion

In vivo

Results
Irritation parameter:
other: ICE class
Basis:
other: average values of the evaluation of corneal lesions
Remarks on result:
other: ICE class II
Irritant / corrosive response data:
Maximal mean corneal swelling score: +10% corresponding to the ICE class II
Maximal mean corneal opacity score: 0.2, corresponding to the ICE class I
Mean score of fluorescein retention: 0.8, corresponding to the ICE class II
Other effects:
no morphological effects were noted, whatever the examination time

Any other information on results incl. tables

Study acceptance criteria:

The concurrent negative and the positive control give an Irritancy Classification that falls within non irritant and severe irritant/corrosive classes, respectively.

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The results obtained, under these experimental conditions, enable to conclude that the test item SEPISOL FAST YELLOW MG-DPG must not be classified R41 "Risk of serious damage to eyes", according to the criteria for the classification, packaging and labelling of dangerous substances in compliance with the E.E.C. Directives 67/548, 2001/59 and 99/45. The symbol "Xi" and the danger label "irritant" are not required.

In accordance with the Regulation (EC) No. 1272/2008, the test item must not be classified in category l "irreversible effects on the eye". The signal word "Danger" and hazard statement H318 "Causes serious eye damage" are not required.
Executive summary:

The aim of the study was to evaluate the possible ocular corrosive or severe irritating effects of the test item after administration on enucleated chickeh eyes.

The test item SEPISOL FAST YELLOW MG-DPG was applied, as supplied, at the dose of 30 mg, to 3 enucleated chicken eyes, during l0 seconds. Then the eyes w ere rinsed twice with l0 mL of physiological saline. As there was remaining test item, the eyes w ere rinsed again twice with l0 mL of physiological saline. Test item remained on the cornea of one eye during all the study and on another eye up to the observation time 2 hours.

Three eyes were treated in the same manner (except for a rinse twice with l0 mL of physiological saline) with a positive control and one eye with a negative control. Damages by the test substance were assessed by determination of corneal swelling, opacity, and fluorescein retention at 30, 75, 120, 180 and 240 minutes post-dose.

The experimental protocol was established in accordance with the O.E.C.D. guideline No 438 adopted 07 September 2009 and the test method B.48 - Commission Regulation (EU) No. 11521201 O dated 08 December 201 O (EU Journal L324) - ATP Council regulation No 440/2008 of 30 May 2008 (E. U Journal Ll42).

The ocular reactions observed in eyes treated with the test item were slight to moderate:

- maximal mean score of corneal opacity: 0.2, corresponding to the ICE class I;

- mean score of fluorescein retention: 0.8, corresponding to the ICE class II;

- maximal mean corneal swelling: +l 0%, corresponding to the ICE class II.

The combination of the three endpoints for the positive control, l0% sodium hydroxide, was 3 x IV. Therefore, the positive control is classified as corrosive/severe irritant, as expected.

The combination of the three endpoints for the negative control, physiological saline solution, was 3 x I. Therefore, the negative control is classified as non corrosive/severe irritant, as expected.

The results obtained, under these experimental conditions, enable to conclude that the test item SEPISOL FAST YELLOW MG-DPG must not be classified R41 "Risk of serious damage to eyes", according to the criteria for the classification, packaging and labelling of dangerous substances in compliance with the E.E.C. Directives 67/548, 2001/59 and 99/45. The symbol "Xi" and the danger label "irritant" are not required.

In accordance with the Regulation (EC) No. 1272/2008, the test item must not be classified in category l "irreversible effects on the eye". The signal word "Danger" and hazard statement H318 "Causes serious eye damage" are not required.