Reaction mass of hexadecyl acrylate and octadecyl acrylate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2012-05-31 to 2012-03-29

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

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)

Version / remarks:

September 07, 2009

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)

Version / remarks:

Commission Regulation (EU) No. 1152/2010 of 8 December 2010

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

BASF SE

Test material

Test material information

Constituent 1

Reference substance name:

2-Propenoic acid, C16-18-alkyl esters

EC Number:

292-060-1

EC Name:

2-Propenoic acid, C16-18-alkyl esters

Cas Number:

90530-21-5

IUPAC Name:

2-Propenoic acid, C16-18-alkyl esters

Test material form:

other: solid

Test animals / tissue source

Species:

other: in vitro (bovine eyes)

Strain:

other: in vitro

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Isolated bovine cornea: The test system (target tissue) 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).
- Source: Schlachthof Bensheim, Am Schlachthof 7-9, 64625 Bensheim
- Age at study initiation: minimum 12 months, maximum 60 months

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 750 µL undiluted test substance (after heating at ca. 30 °C)

Duration of treatment / exposure:

10 minutes

Duration of post- treatment incubation (in vitro):

2-hour post-incubation period

Number of animals or in vitro replicates:

3 corneas

Details on study design:

Corneal opacity was measured quantitatively as the amount of light transmission through the cornea. Permeability was measured quantitatively as the amount of sodium fluorescein dye that passes across the full thickness of the cornea. Both measurements were used to calculate an In Vitro Irritancy Score of the test substance relative to the control corneas.
Corneas free of defects (opacity, scratches, pigmentation etc.) were dissected with a 2 to 3 mm rim of sclera. Isolated corneas were mounted in specially designed corneal holders that consists of anterior and posterior chambers. Both chambers were filled to excess with prewarmed
Eagles’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 show macroscopic tissue damage or an opacity value < 507 opacity units were discarded. Corneas with opacity values close to the median value of all corneas were selected as negative control (NC). The remaining corneas were then distributed into treatment and positive control 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 uL of the undiluted liquid test substance was applied into the anterior chamber using a pipette. Control tissues were concurrently applied into the anterior chamber with 750 uL of de-ionized water (negative control, NC) or with 750 μL of 1% (w/v) solution of sodium hydroxide in deionized water (positive control, PC) 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.

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

cornea opacity score

Value:

1.1

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Results 2

Irritation parameter:

other: Permeability

Value:

0.002

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Results 3

Irritation parameter:

in vitro irritation score

Value:

1.2

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

Test Substance	Mean Opacity Value	Mean Permeability Value	In Vitro Irritation Score
12/0073-1	1.1	0.002	1.2
NC	2.8	-0.010	2.6
PC	121.7	3.204	169.7

Applicant's summary and conclusion

Interpretation of results:

other: not severely eye damaging