Registration Dossier

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018

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
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Specific details on test material used for the study:
TEST MATERIAL NAME (as stated in study report): Internediate F37

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Valtris Specialty Chemicals Ltd. / Batch/Lot No. W042272
- Expiration date of the lot/batch: November 21, 2019
- Purity: 84.5%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature. Containers kept tightly closed in a dry, cool and well ventilated place.

Test animals / tissue source

Species:
cattle
Strain:
not specified
Details on test animals or tissues and environmental conditions:
Test System : Isolated Cornea from the eyes of freshly slaughtered cattle
Source : Deonar Abattoir slaughter house, Mumbai, Maharashtra
Bovine Age : Between 1 to 5 years (age of the animals was determined based on the teeth count and horn ring count along with Horizontal Diameter of Corneas and Central Corneal Thickness)
Transportation Condition : Transported (in a sealed plastic container) under cold condition in Hanks’ Balanced Salt Solution containing antibiotics [e.g., penicillin at 100 IU/mL and streptomycin at 100 μg/mL]

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): 750 µL
- Concentration (if solution): undiluted
Duration of treatment / exposure:
approximately 10 minutes ± 30 seconds
Duration of post- treatment incubation (in vitro):
approximately 2 hours ± 10 minutes
Number of animals or in vitro replicates:
three corneas per test group
Details on study design:
SELECTION AND PREPARATION OF CORNEAS
The eyes were used within 24 hours from slaughter. Eyes were examined prior to use. Corneas from eyes free of visible defects were used. Corneas that have opacity lesser than seven opacity units or equivalent for the opacitometer were used in the study.

Corneas, free from defects, were dissected to a 2 to 3 mm rim and were transferred to container containing Hanks’ Balanced Salt Solution containing antibiotics [e.g., penicillin at 100 IU/mL and streptomycin at 100 μg/mL].

Selected corneas were mounted on the corneal holders with the endothelial side against the O-ring of the posterior half of the holder. The anterior half of the holder was then placed on the top of the cornea and fixed in place. Both chambers were then filled to excess with pre-warmed phenol red free Eagle's Minimum Essential Medium (EMEM) (posterior chamber first to return the cornea to its natural concave position), ensuring no bubbles were present within the holders. The device was then equilibrated at 32 ± 1°C for at least one hour to allow the corneas to equilibrate with the medium and to achieve normal metabolic activity, to the extent possible. Following the equilibration period, the medium was removed from both the chambers and fresh pre-warmed phenol red free EMEM was added to both chambers and baseline opacity readings were taken for each cornea.

REMOVAL OF TEST SUBSTANCE
At the end of exposure period, the test item, positive control and negative control were removed from their respective anterior chamber and the corneal epithelium washed until no visual evidence of test item, positive control or negative control was observed using EMEM (containing phenol red).

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: light transmission
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of [UV/VIS spectrophotometry

SCORING SYSTEM: In Vitro Irritancy Score (IVIS)

DECISION CRITERIA: per test guideline

Results and discussion

In vitro

Results
Irritation parameter:
in vitro irritation score
Remarks:
IVIS
Value:
0.53
Negative controls validity:
valid
Positive controls validity:
valid
Other effects / acceptance of results:
DEMONSTRATION OF TECHNICAL PROFICIENCY: yes

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Remarks:
Not classified by CLP Criteria
Conclusions:
Under guideline (OECD 437) test conditions, the mean IVIS score for Intermediate F37 in the bovine corneal opacity and permeabilty (BCOP) test was 0.53. Based on these results, Intermediate F37 is not classified for eye irritation or serious eye damage.