Registration Dossier

Diss Factsheets

Administrative data

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

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report date:
2017

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)
Version / remarks:
2013
Deviations:
no
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
-
EC Number:
439-510-7
EC Name:
-
Cas Number:
149048-48-6
Molecular formula:
Constituent 1: C14H26N2O3Si Constituent 2: C13H22N2O2Si
IUPAC Name:
1-(3-{13-[3-(aminomethyl)phenyl]-6,6,8,8-tetramethoxy-7-oxa-2,12-diaza-6,8-disilatridecan-1-yl}phenyl)methanamine; 1-[3-({[3-(trimethoxysilyl)propyl]amino}methyl)phenyl]methanamine
Test material form:
liquid
Specific details on test material used for the study:
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature container flushed with nitrogen
- Stability under test conditions: stable
- Solubility and stability of the test substance in the solvent/vehicle: The test material was applied undiluted.
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: not applicable

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test material was applied undiluted.
- Preliminary purification step (if any): No correction was made for the purity/composition of the test item.
- Final dilution of a dissolved solid, stock liquid or gel: not applicable
- Final preparation of a solid: not applicable

Test animals / tissue source

Species:
cattle
Strain:
not specified
Details on test animals or tissues and environmental conditions:
SOURCE OF COLLECTED EYES
- Source: Vitelco, 's Hertogenbosch, The Netherlands
- Number of animals:
- Characteristics of donor animals (e.g. age, sex, weight): young cattle
- Storage, temperature and transport conditions of ocular tissue (e.g. transport time, transport media and temperature, and other conditions): Eyes were collected and transported in physiological saline in a suitable container under cooled conditions.
- Time interval prior to initiating testing:
- indication of any existing defects or lesions in ocular tissue samples: The eyes were checked for unacceptable defects, such as opacity, scratches, pigmentation and neovascularization by removing them from the physiological saline and holding them in the light. Those exhibiting defects were discarded.
- Indication of any antibiotics used: none specified

Test system

Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 750µl
- Concentration (if solution): not applicable

Duration of treatment / exposure:
10 +/- 1 minutes at 32 +/- 1°C
Duration of post- treatment incubation (in vitro):
120 +/- 10 minutes at 32 +/- 1°C
Number of animals or in vitro replicates:
Three corneas per treatment group.
Details on study design:
SELECTION AND PREPARATION OF CORNEAS: The eyes were checked for unacceptable defects, such as opacity, scratches, pigmentation and neovascularization by removing them from the physiological saline and holding them in the light. Those exhibiting defects were discarded. The corneas were incubated with cMEM for the minimum of 1 hour at 32 +/- 1°C.

QUALITY CHECK OF THE ISOLATED CORNEAS: After the incubation period, the medium was removed from both compartments and replaced with fresh cMEM. Opacity determinations were performed on each of the corneas using an opacitometer. The opacity of each cornea was read against a cMEM filled chamber, and the initial opacity reading thus determined was recorded. Corneas that had an initial opacity reading higher than 7 were not used. Three corneas were selected at random for each treatment group.

NUMBER OF REPLICATES: Three corneas per group

NEGATIVE CONTROL USED: Yes, physiological saline

SOLVENT CONTROL USED (if applicable): not applicable

POSITIVE CONTROL USED: Yes, ethanol

APPLICATION DOSE AND EXPOSURE TIME: 750µl for 10 +/- 1 minutes at 32 +/- 1°C

TREATMENT METHO: not specified

POST-INCUBATION PERIOD: No

REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period: After the incubation the solutions were removed and the epithelium was washed with MEM with phenol red (Earle’s Minimum Essential Medium) and thereafter with cMEM
- POST-EXPOSURE INCUBATION: 120 +/- 10 minutes at 32 +/- 1°C

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: diminution of light passing through the cornea
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of microtiter plate reader (OD490)
- Others (e.g, pertinent visual observations, histopathology):

SCORING SYSTEM: In Vitro Irritancy Score (IVIS)

DECISION CRITERIA: The decision criteria as indicated in the TG was used.

Results and discussion

In vitro

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Run / experiment:
treatment with 100% test material for 10 min
Value:
> 186 - <= 304
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Remarks:
ranged from -2.2 to -1.0
Positive controls validity:
valid
Remarks:
ranged from 51 to 71
Remarks on result:
positive indication of irritation
Irritation parameter:
in vitro irritation score
Run / experiment:
treatment with 100% test material for 10 min
Value:
228
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
positive indication of irritation
Other effects / acceptance of results:
OTHER EFFECTS:
- Visible damage on test system: none reported

DEMONSTRATION OF TECHNICAL PROFICIENCY: 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 61 and within two standard deviations of the current historical positive control mean (Appendix 3). It was therefore concluded that the test conditions were adequate and that the test system functioned properly.

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Yes, within historical control ranges
- Acceptance criteria met for positive control: Yes, within historical control ranges
- Range of historical values if different from the ones specified in the test guideline: Not different

Applicant's summary and conclusion

Interpretation of results:
Category 1 (irreversible effects on the eye) based on GHS criteria
Conclusions:
In the Bovine Corneal Opacity and Permeability test, conducted according to an appropriate OECD guideline and in compliance with GLP, the test material was reported to induce an IVIS score greater than 55 and to be irritating to bovine corneas.