4-fluoro-1,3-dioxolan-2-one

Eye irritation

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:

2018

Report date:

2018

Materials and methods

Test guideline

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)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

483-360-5

EC Name:

-

Cas Number:

114435-02-8

Molecular formula:

C3H3FO3

IUPAC Name:

4-fluoro-1,3-dioxolan-2-one

Test material form:

solid

Test animals / tissue source

Species:

cattle

Strain:

not specified

Details on test animals or tissues and environmental conditions:

SOURCE OF COLLECTED EYES
- Source: Slaughterhouse (Kumamoto Chuou Meat Centre Co., Ltd, 548, Subayashi, Toyono-machi, Uki-shi, Kumamoto 861-4307, Japan
- Number of animals: Fifteen eyes excised from animals
- Characteristics of donor animals (e.g. age, sex, weight): 23 to 25 months old.
- Storage, temperature and transport conditions of ocular tissue (e.g. transport time, transport media and temperature, and other conditions): Transported in a cooler with Hanks' balanced salt solution.
- Time interval prior to initiating testing: Used on day of receipt
- indication of any existing defects or lesions in ocular tissue samples: There were no eyes exhibiting defects.
- Indication of any antibiotics used: None

Test system

Vehicle:

physiological saline

Remarks:

Isotonic sodium chloride solution

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 750 µL (test substance heated so tested in liquid form)
- Concentration (if solution): NA used as supplied

VEHICLE
- Amount(s) applied (volume or weight with unit): 750 µL
- Concentration (if solution): NA used as supplied

Duration of treatment / exposure:

Corneas were incubated for 10 minutes

Duration of post- treatment incubation (in vitro):

2 hours in a low temperature incubator

Number of animals or in vitro replicates:

Three corneas were selected for each group

Details on study design:

SELECTION AND PREPARATION OF CORNEAS : The corneal holder, MEM and cMEM were warmed in a water batch until use. Corneas were dissected with a 2 to 3 mm rim of sclera remaining. Isolated corneas were mounted in corneal holders with anterior and posterior compartments which interface with epithelial and endothelial slides of the cornea, respectively. Both compartments were filled with cMEM. Corneas were incubated for 1 hour in a low temperature incubator (set to 32°C). 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 was calculated according to the equation in the test guideline. Corneas that had an initial opacity reading higher than 7 were not used. Nine corneas were chosen for the experiment excluding those with high opacity.

QUALITY CHECK OF THE ISOLATED CORNEAS: The eyes were checked for unacceptable defects such as opacity, scratches and neovascularization before use.

NUMBER OF REPLICATES : Three corneas were selected for each group

NEGATIVE CONTROL USED: Physiological saline

POSITIVE CONTROL USED : N,N-dimethylformamide

APPLICATION DOSE AND EXPOSURE TIME : 700 µL (10 minutes exposure time)

TREATMENT METHOD: closed chamber

POST-INCUBATION PERIOD: After the exposure incubation corneas were washed three times with MEM after which the corneas were rinsed a final time with cMEM. These were then incubated for a further 2 hours in a low temperature incubation (set at 32°C).

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

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: Opacity determinations were performed using an opacitometer
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of microtiter plate reader (OD490)
- Others (e.g, pertinent visual observations, histopathology): Each cornea was visually observed (e.g. tissue peeling, residual test substance, non-uniform opacity patterns).

SCORING SYSTEM: The final opacity for each cornea was calculated by subtracting the initial opacity reading from the post-treatment reading. The mean of the final opacity value of each treatment group was calculated. To calculate the IVIS score, values obtained by subtracting the mean final opacity of the negative control from that of the test substance or positive control were used for the calculation. For permeability each value (OD490) was corrected for the background value (OD490 of cMEM only).

DECISION CRITERIA: Decision criteria as per the test guideline used.

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Run / experiment:

2

Value:

>= 10.9 - <= 11.3

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Remarks:

IVIS scores of 0.2 to 1.1

Positive controls validity:

valid

Remarks:

IVIS scores of 92.9 to 99.7

Other effects / acceptance of results:

OTHER EFFECTS:
- Visible damage on test system: None noted

DEMONSTRATION OF TECHNICAL PROFICIENCY: Results as expected based on historical control data for BCOP studies

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Yes
- Acceptance criteria met for positive control: Yes
- Range of historical values if different from the ones specified in the test guideline: NA

Applicant's summary and conclusion

Interpretation of results:

study cannot be used for classification

Conclusions:

Since the test substance induced an IVIS score > 3 and < 55, no prediction could be made for eye irritation or serious eye damage.