Registration Dossier

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Test accoding to GLP and the study procedures described in this report were based on the most recent OECD and EC guidelines.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report Date:
2015

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)
Qualifier:
according to
Guideline:
EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
Identification: V173520
Appearance: White powder (determined by WIL Research Europe)
Batch: AN
Purity/Composition: Not indicated
Test substance storage: At room temperature
Stable under storage conditions until: 20 April 2017 (expiry date)

Purity/composition correction factor: No correction factor required
Test substance handling: No specific handling conditions required
Chemical name (IUPAC), synonym or trade name: 1,1,2,2-tetramethyl-1H-Benz(e)indolium 4- methylbenzenesulfonate
CAS Number: 141914-99-0
Molecular formula: C23H25NO3S
Molecular weight: 395.53
pH (1% in water, indicative range) : 4.1 – 4.03 (determined by WIL Research Europe B.V. )
Solubility in vehicle: Physiological saline: Not indicated
Stability in vehicle: Physiological saline: Not indicated

Test animals / tissue source

Species:
other: Bovine eyes
Details on test animals or tissues and environmental conditions:
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.
Eyes were collected and transported in physiological saline in a suitable container under cooled conditions.

Test system

Vehicle:
physiological saline
Remarks:
A 20% (w/v) solution of V173520 was prepared in physiological saline (Eurovet Animal Health, Bladel, The Netherlands).
Controls:
yes
Amount / concentration applied:
V173520 was tested in a 20% (w/v) solution.
Duration of treatment / exposure:
240 min
Details on study design:
Treatment of corneas and opacity measurements

750 μl of either the negative control, positive control (20% (w/v) Imidazole solution) or 20% (w/v) 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 ± 1°C. 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 Life Technologies). Possible pH effects of the test substance on the corneas were recorded. Each cornea was inspected visually for dissimilar opacity patterns. The medium in the posterior compartment was removed and both compartments were refilled with fresh cMEM and the opacity determinations were performed.

Results and discussion

Results of in vivo study
Irritation parameter:
other: In vitro irritancy scores
Basis:
other: range
Time point:
other: after 240 minutes
Score:
394 - 415
Max. score:
415
Irritant / corrosive response data:
The corneas treated with V173520 showed opacity values ranging from 383 to 385 and permeability values ranging from 0.720 to 2.018. The corneas were black after the 240 minutes of treatment with V173520. No pH effect of the test substance was observed on the rinsing medium. Hence, the in vitro irritancy scores ranged from 394 to 415 after 240 minutes of treatment with V173520.

Applicant's summary and conclusion

Interpretation of results:
Category 1 (irreversible effects on the eye) based on GHS criteria
Remarks:
Migrated information
Conclusions:
Since V173520 induced an IVIS ≥ 55, it is concluded that V173520 induces serious eye damage in the Bovine Corneal Opacity and Permeability test under the experimental conditions described in this report and should be classified category 1 according to the Globally Harmonized System of Classification and Labeling of Chemicals (GHS) of the United Nations.