Registration Dossier

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
Jul 2019
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2019
Report date:
2019

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
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)
Version / remarks:
version Oct 2017
Qualifier:
according to guideline
Guideline:
EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
3a,4,7,7a-tetrahydro-1H-indene
EC Number:
221-260-3
EC Name:
3a,4,7,7a-tetrahydro-1H-indene
Cas Number:
3048-65-5
Molecular formula:
C9H12
IUPAC Name:
3a,4,7,7a-tetrahydro-1H-indene
Constituent 2
Chemical structure
Reference substance name:
4-vinylcyclohexene
EC Number:
202-848-9
EC Name:
4-vinylcyclohexene
Cas Number:
100-40-3
Molecular formula:
C8H12
IUPAC Name:
4-vinylcyclohexene
Constituent 3
Chemical structure
Reference substance name:
Cyclopentadiene
EC Number:
208-835-4
EC Name:
Cyclopentadiene
Cas Number:
542-92-7
Molecular formula:
C5H6
IUPAC Name:
cyclopenta-1,3-diene
Constituent 4
Chemical structure
Reference substance name:
cis-1,2-Divinylcyclobutane
Cas Number:
2422-85-7
Molecular formula:
C8H12
IUPAC Name:
cis-1,2-Divinylcyclobutane
Constituent 5
Chemical structure
Reference substance name:
Ethylbenzene
EC Number:
202-849-4
EC Name:
Ethylbenzene
Cas Number:
100-41-4
Molecular formula:
C8H10
IUPAC Name:
ethylbenzene
Constituent 6
Chemical structure
Reference substance name:
5-vinylnorborn-2-ene
EC Number:
221-259-8
EC Name:
5-vinylnorborn-2-ene
Cas Number:
3048-64-4
Molecular formula:
C9H12
IUPAC Name:
5-vinylbicyclo[2.2.1]hept-2-ene
Constituent 7
Chemical structure
Reference substance name:
Cycloocta-1,5-diene
EC Number:
203-907-1
EC Name:
Cycloocta-1,5-diene
Cas Number:
111-78-4
Molecular formula:
C8H12
IUPAC Name:
cycloocta-1,5-diene
Constituent 8
Chemical structure
Reference substance name:
3a,4,7,7a-tetrahydro-4,7-methanoindene
EC Number:
201-052-9
EC Name:
3a,4,7,7a-tetrahydro-4,7-methanoindene
Cas Number:
77-73-6
Molecular formula:
C10H12
IUPAC Name:
3a,4,7,7a-tetrahydro-4,7-methanoindene,
Constituent 9
Chemical structure
Reference substance name:
3a,4,7,7a-tetrahydro-4-methyl-1H-indene
EC Number:
231-021-5
EC Name:
3a,4,7,7a-tetrahydro-4-methyl-1H-indene
Cas Number:
7413-02-7
Molecular formula:
C10H14
IUPAC Name:
4-methyl-3a,4,7,7a-tetrahydro-1H-indene
impurity 1
Chemical structure
Reference substance name:
Combination of: 6-(cyclohex-3-en-1-yl)bicyclo[2.2.1]hept-2-ene 2-vinyl-1,2,3,4,4a,5,8,8a-octahydro-1,4-methanonaphthalene 6-vinyl-1,4,4a,5,6,7,8,8a-octahydro-1,4-methanonaphthalene 2-vinyl-1,2,3,4,4a,5,8,8a-octahydro-1,4:5,8-dimethanonaphthalene 2,2'-bi(bicyclo[2.2.1]heptane-5,5'-diene) 3a,4,4a,5,8,8a,9,9a-octahydro-1H-5,8-methanocyclopenta[b]naphthalene 4,4a,4b,5,8,8a,9,9a-octahydro-1H-1,4-methanofluorene
Molecular formula:
C13H18 to C14H18
IUPAC Name:
Combination of: 6-(cyclohex-3-en-1-yl)bicyclo[2.2.1]hept-2-ene 2-vinyl-1,2,3,4,4a,5,8,8a-octahydro-1,4-methanonaphthalene 6-vinyl-1,4,4a,5,6,7,8,8a-octahydro-1,4-methanonaphthalene 2-vinyl-1,2,3,4,4a,5,8,8a-octahydro-1,4:5,8-dimethanonaphthalene 2,2'-bi(bicyclo[2.2.1]heptane-5,5'-diene) 3a,4,4a,5,8,8a,9,9a-octahydro-1H-5,8-methanocyclopenta[b]naphthalene 4,4a,4b,5,8,8a,9,9a-octahydro-1H-1,4-methanofluorene
Constituent 10
Chemical structure
Reference substance name:
4,4a,4b,5,8,8a,9,9a-octahydro-1H-1,4-methanofluorene OR 4,4a,4b,5,8,8a,9,9a-octahydro-1H-1,4:5,8-dimethanofluorene
Molecular formula:
C14H18 to C15H18
IUPAC Name:
4,4a,4b,5,8,8a,9,9a-octahydro-1H-1,4-methanofluorene OR 4,4a,4b,5,8,8a,9,9a-octahydro-1H-1,4:5,8-dimethanofluorene
Constituent 11
Chemical structure
Reference substance name:
3a,4,4a,5,8,8a,9,9a-octahydro-4,9:5,8-dimethano-1H-benz[f]indene
EC Number:
230-512-1
EC Name:
3a,4,4a,5,8,8a,9,9a-octahydro-4,9:5,8-dimethano-1H-benz[f]indene
Cas Number:
7158-25-0
Molecular formula:
C15H18
IUPAC Name:
3a,4,4a,5,8,8a,9,9a-octahydro-1H-4,9:5,8-dimethanocyclopenta[b]naphthalene
Constituent 12
Reference substance name:
Tetramers of cyclopentadiene and 1,3-butadiene
Molecular formula:
C16H24 to C20H24
IUPAC Name:
Tetramers of cyclopentadiene and 1,3-butadiene
Test material form:
liquid
Details on test material:
Light brown liquid.
Batch number 776809

Test animals / tissue source

Species:
cattle
Strain:
not specified
Details on test animals or tissues and environmental conditions:
SOURCE OF COLLECTED EYES
- Source: Adult cattle from local abattoir
- Number of animals: not specified
- Characteristics of donor animals (e.g. age, sex, weight): 12-60 months old
- Storage, temperature and transport conditions of ocular tissue (e.g. transport time, transport media and temperature, and other conditions): Eye transported same day in Hanks' balanced salt solution (HBBS) in ice packs
- Time interval prior to initiating testing: Corneas prepared immediately on arrival
- indication of any existing defects or lesions in ocular tissue samples: Examined macroscopically and only corneas free of damage used.
- Indication of any antibiotics used: HBBS augmented with penicillin and streptomycin.

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): 0.75mL
Duration of treatment / exposure:
10 mins
Duration of post- treatment incubation (in vitro):
After rinsing, 120 mins at 32C.
Number of animals or in vitro replicates:
3
Details on study design:
SELECTION AND PREPARATION OF CORNEAS : All eyes were macroscopically examined before and after dissection. Only corneas free of damage were used. The cornea from each selected eye was removed leaving a 2 to 3 mm rim of sclera to facilitate handling. The iris and lens were peeled away from the cornea. The isolated corneas were immersed in a dish containing HBSS until they were mounted in Bovine Corneal Opacity and Permeability (BCOP) holders. The anterior and posterior chambers of each BCOP holder were filled with complete Eagle’s Minimum Essential Medium (EMEM) without phenol red and plugged. The holders were incubated at 32 ± 1 ºC for 60 minutes. At the end of the incubation period each cornea was examined for defects. Only corneas free of damage were used.

NUMBER OF REPLICATES : 3

NEGATIVE CONTROL USED : Yes, 0.9% saline

POSITIVE CONTROL USED : Yes, pure ethanol

APPLICATION DOSE AND EXPOSURE TIME 0.75mL for 10 mins

REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period: 3

- POST-EXPOSURE INCUBATION: 120 mins at 32C.

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: not specified
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of microtiter plate reader
- Other: corneas retained for histopathology but this was not performed.

SCORING SYSTEM: In Vitro Irritancy Score (IVIS)

DECISION CRITERIA: As per UN GHS (<=3 not classified, >55 category 1)

Results and discussion

In vitro

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Run / experiment:
Average of 3 corneas
Value:
ca. 1.3
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Remarks:
corrected score
Irritation parameter:
in vitro irritation score
Run / experiment:
Average of 3 corneas
Value:
ca. 1.5
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Other effects / acceptance of results:
OTHER EFFECTS:
- Visible damage on test system: No

DEMONSTRATION OF TECHNICAL PROFICIENCY:

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
Executive summary:

In a guideline and GLP ex-vivo bovine corneal opacity eye irritation study, the substance THI-TM produced minimal effects on the cattle corneas used. The results indicated that the substance does not need to be classified as an eye irritant