C10-14-alkyl-1-(ethenyloxy)-

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

30 December 2017 to 03 January 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Reference

Name:

Unnamed

Type:

Constituent

Test material form:

liquid

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material:
Kyoeisha chemical co., Bx 7080101
- Expiration date of the lot/batch: August 01, 2019
- Purity test date: no data

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature (Ambient), Container was kept tightly closed and away from heat or sunlight
- Stability under test conditions: no data
- Solubility and stability of the test substance in the solvent/vehicle: - Reactivity of the test substance with the solvent/vehicle of the cell culture medium:

Test animals / tissue source

Species:

rat

Strain:

Wistar

Details on test animals or tissues and environmental conditions:

SOURCE OF COLLECTED EYES
- Source: Deonar Abattoir slaughter house, Mumbai, Maharashtra
- Number of animals: no data
- Characteristics of donor animals (e.g. age, sex, weight): 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)
- Storage, temperature and transport conditions of ocular tissue (e.g. transport time, transport media and temperature, and other conditions): 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]
- Time interval prior to initiating testing: The eyes were used within 24 hours from slaughter.
- indication of any existing defects or lesions in ocular tissue samples: 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.
- Indication of any antibiotics used: 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:

not specified

Controls:

yes, concurrent positive control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit):
- Concentration (if solution): 1% aqueous solution in distilled water at room temperature

VEHICLE
- Amount(s) applied (volume or weight with unit): none
- Concentration (if solution):
- Lot/batch no. (if required):
- Purity:

Duration of treatment / exposure:

Corneas were exposed for approximately 10 minutes ± 30 seconds at 32 ± 1°C.

Duration of post- treatment incubation (in vitro):

2 hours ± 10 minutes at 32 ± 1ºC.

Number of animals or in vitro replicates:

12

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.

QUALITY CHECK OF THE ISOLATED CORNEAS ; 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.

NUMBER OF REPLICATES ; 3

NEGATIVE CONTROL USED ; yes

SOLVENT CONTROL USED (if applicable)

POSITIVE CONTROL USED ; yes

APPLICATION DOSE AND EXPOSURE TIME ; 750 μL 10 minutes ± 30 seconds at 32 ± 1 °C.

TREATMENT METHOD: closed chamber

POST-INCUBATION PERIOD: yes. 2 hours ± 10 minutes at 32 ± 1ºC.

REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period: one
- POST-EXPOSURE INCUBATION: 2 hours ± 10 minutes at 32 ± 1ºC.

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: Corneal opacity is measured quantitatively, as the amount of light transmission through the cornea.
- Corneal permeability: Permeability is measured quantitatively as the amount of sodium fluorescein dye that passes across the full thickness of the cornea, as detected in the medium in the posterior chamber. The test item is applied to the epithelial surface of the cornea by addition to the anterior chamber of the corneal holder.
- Others (e.g, pertinent visual observations, histopathology): (please specify)

SCORING SYSTEM: In Vitro Irritancy Score (IVIS) IVIS = mean opacity value + (15 x mean permeability OD490 value)

DECISION CRITERIA: please specify if the decision criteria as indicated in the TG was used.

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

in vitro irritation score

Run / experiment:

17

Value:

1.03

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Results 2

Irritation parameter:

in vitro irritation score

Run / experiment:

23

Value:

-0.77

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Results 3

Irritation parameter:

in vitro irritation score

Run / experiment:

25

Value:

2.89

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Results 4

Irritation parameter:

cornea opacity score

Run / experiment:

17

Value:

1.58

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Results 5

Irritation parameter:

cornea opacity score

Run / experiment:

23

Value:

-0.95

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Results 6

Irritation parameter:

cornea opacity score

Run / experiment:

25

Value:

2.38

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The mean IVIS score for the corneas treated with C10-C14 alkyl ether was found to be 1.05. Based on the results of this study, an indication of the classification for C10-C14 alkyl ether is as follows: not classified as irritating to eyes.