2,3-dihydroxypropyl methacrylate

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:

2017

Report date:

2016

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 (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

2,3-dihydroxypropyl methacrylate

EC Number:

227-642-6

EC Name:

2,3-dihydroxypropyl methacrylate

Cas Number:

5919-74-4

Molecular formula:

C7H12O4

IUPAC Name:

2,3-dihydroxypropyl 2-methylprop-2-enoate

Test animals / tissue source

Species:

cattle

Strain:

not specified

Details on test animals or tissues and environmental conditions:

BCOP
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 Mannheim, Schlachthofstr. 21, 68165 Mannheim, Germany

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Details on study design:

BCOP:
Several test substances were tested in parallel within the present test (test no. 209) using the same control corneas (NC and PC).
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 with fresh prewarmed 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 < 548 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, NC and PC) consisted of 3 corneas. Before application, the medium in the anterior chamber was removed using a syringe. 750 μL of the undiluted liquid test substance was applied into the anterior chamber using a pipette. For the control tissues 750 μL of de-ionized water (negative control, NC) or 750 μL of 100% ethanol / 100% dimethylformamide (positive controls, PC 1 and PC 2), were applied into the anterior chamber 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, NC and PC were then removed from the anterior chamber 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 further 2 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 observed visually 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 min in a horizontal position at about 32 °C.
The amount of sodium fluorescein that permeated through the corneas into the posterior chamber was measured spectrophotometrically. Three aliquots per cornea were transferred to a 96-well microtiter plate and the optical density (OD490) was determined.

Controls BCOP
Negative control (NC): De-ionized water
Positive control (PC): 100% ethanol (PC 1) for liquid test substances and surfactants; 100% dimethylformamide (PC 2) was used additionally to increase the historic database of the PC.

Results and discussion

In vitro

Results

Remarks on result:

other: The mean IVIS of the test-substance treated corneas was 27.3.

Any other information on results incl. tables

Result BCOP:

Test substance	Mean Opacity Value	Mean Permeability Value	Mean In Vitro Irritancy Score
16/0064-1	19.6	0.510	27.3
NC	5.6	0.002	5.6
PC 1	20.0	1.020	35.3
PC 2	90.0	0.670	100.1

Summary table for turnkey test strategy evaluation:

Test Method	Test Result	Test Evaluation	Evaluation Test Strategy
BCOP Test	The mean IVIS of the test-substance treated corneas was 27.3.	Not identified as corrosive or severe irritant	Ocular irritant
EpiOcular	Mean viability of the test-substance treated tissues was 5.5%.	Irritant

 

Applicant's summary and conclusion

Interpretation of results:

other: not identified as corrosive or severe irritant

Conclusions:

Based on the results for BCOP and applying the evaluation criteria, GMMA was not identified as corrosive or severe irritant under the test conditions chosen.

The irritation potential of the test substance was assessed in the BCOP assay. Application of the test substance to bovine corneae resulted in a calculated mean IVIS of 27.3. Thus, the test substance is considered to be non-corrosive, but no prediction on the irritation potential can be made and further evaluation and/or data generation is required.