2-Propenoic acid, (2,2-dimethyl-1,3-dioxolan-4-yl)methyl ester

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

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 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)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

(2,2-dimethyl-1,3-dioxolan-4-yl)methyl prop-2-enoate

EC Number:

814-155-4

Cas Number:

13188-82-4

Molecular formula:

C9H14O4

IUPAC Name:

(2,2-dimethyl-1,3-dioxolan-4-yl)methyl prop-2-enoate

Test material form:

liquid

Test animals / tissue source

Details on test animals or tissues and environmental conditions:

Isolated bovine cornea: The test system is the isolated bovine cornea. Bovine eyes are obtained as a by-product of freshly slaughtered cattle (age of the animals: minimum 12 months, maximum 60 months) Schlachthof Alzey, Germany.

Corneas free of defects (opacity, scratches, pigmentation etc.) were dissected with a 2 to 3 mm rim of sclera. Isolated corneas were mounted in cornea holders that consists of anterior and posterior chambers. Both chambers were filled to excess with pre-warmed Eagle’s MEM (without phenol red) and then equilibrated in a vertical position at about 32°C for at least 1 hour.
After the equilibration period, the medium in both chambers was replaced by fresh pre-warmed medium and initial corneal opacity readings were taken for each cornea with an opacitometer. Any corneas that showed macroscopic tissue damage or an opacity value < 556 opacity units1 were discarded. The remaining corneas were then distributed into negative control, positive control and treatment groups. Each corneal holder was uniquely identified with a number on the chambers.

Each treatment group (test substance, the NC and the PC) consisted of 3 corneas. Before application, the medium in the anterior chamber was removed by using a syringe. 750 μL undiluted liquid test substance were applied into the anterior chamber by using a pipette. For the control tissues, 750 μL deionized water (negative control, NC) or 750 μL 100% ethanol / 100% dimethylformamide (positive controls, PC1 / PC2) were applied into the anterior chamber by using a pipette. The corneas were incubated in a horizontal position at about 32°C for approximately 10 minutes (liquids and surfactants). The test substance, the NC and the PC were then removed from the anterior chamber by using a syringe and the epithelium was washed at least 3 times with Eagle’s MEM (containing phenol red) and once with Eagle’s MEM (without phenol red). Both chambers were then refilled with fresh Eagle’s MEM (without phenol red).

The corneas were incubated for 2 further hours at about 32°C. After the incubation period, the medium was removed and both chambers were then refilled with fresh Eagle’s MEM.

Before measurement, each cornea was visually observed and observations were recorded. Final corneal opacity readings were taken for each cornea with an opacitometer.

For determination of permeability, the medium in the anterior chamber was replaced by 1 mL sodium fluorescein solution (4 mg/mL for liquid test substances and surfactants) and incubated for 90 ± 5 minutes in a horizontal position at about 32°C. The amount of sodium fluorescein that permeated through the corneas into the posterior chamber was spectrophotometrically measured. Three aliquots per cornea were transferred to a 96-well microtiter plate and the optical density (OD490) was determined.

Negative control (NC): Deionized water
Positive control (PC): 100% ethanol (PC1) / 100% dimethylformamide (PC2) for liquid test substances and surfactants

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Run / experiment:

mean

Value:

6.5

Any other information on results incl. tables

Result BCOP:

Test substance	Mean Opacity Value	Mean Permeability Value	Mean In Vitro Irritancy Score
18/0245-1	6.5	0.002	6.5
NC	4.3	0.001	4.4
PC1	24.8	0.751	36.1
PC2	102.1	0.389	107.9

Summary of individual test results and test strategy evaluation

Test Method	Test Result	Test Evaluation	Evaluation Test Strategy
BCOP Test	The mean IVIS of the corneas treated with the test substance was 6.5.	Not identified as corrosive or severe irritant	Ocular irritant
EpiOcular	The final relative mean Ocular irritant viability of the tissues treated with the test substance was 32.5%.	Irritant

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

Based on the results of the BCOP and EpiOcular Tests and by applying the evaluation criteria, it was concluded that Laromer IPGA shows an eye irritation potential in the in vitro eye irritation test strategy under the test conditions chosen.