Fatty acids, tall-oil, polymers with glycidyl neodecanoate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From June 29, 2015 to July 8, 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

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

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: OECD Guideline for the testing of chemicals, series on testing and assessment no. 160: “Guidance document on “the bovine corneal opacity and permeability (bcop) and isolated chicken eye (ice) test methods.

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

2-hydroxy-3-[(2,2,3,5-tetramethylhexanoyl)oxy]propyl (9Z)-octadec-9-enoate; 2-hydroxy-3-[(2,2,3,5-tetramethylhexanoyl)oxy]propyl (9Z,12Z)-octadeca-9,12-dienoate; 2-hydroxy-3-[(2,2,3,5-tetramethylhexanoyl)oxy]propyl octadecanoate

EC Number:

617-001-2

Cas Number:

80207-00-7

Molecular formula:

not applicable

IUPAC Name:

2-hydroxy-3-[(2,2,3,5-tetramethylhexanoyl)oxy]propyl (9Z)-octadec-9-enoate; 2-hydroxy-3-[(2,2,3,5-tetramethylhexanoyl)oxy]propyl (9Z,12Z)-octadeca-9,12-dienoate; 2-hydroxy-3-[(2,2,3,5-tetramethylhexanoyl)oxy]propyl octadecanoate

Test material form:

liquid

Remarks:

Brown

Test animals / tissue source

Species:

other: Bos primigenius taurus (fresh bovine corneas)

Details on test animals or tissues and environmental conditions:

- Species: Bos primigenius Taurus (fresh bovine corneas).
- Source: Slaughterhouse Müller Fleisch GmbH, Enzstr. 2-4, 75217 Birkenfeld, Germany.
- Age of cattles: Between 12 and 60 months old.
- Transport of eyes: In Hank’s balanced salt solution (supplemented with 0.01% streptomycin and 0.01% penicillin).

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: Negative control: 0.9% NaCl; positive control: Dimethylformamide, DMF

Amount / concentration applied:

TEST SUBSTANCE
- Amount applied: 750 μL

Duration of treatment / exposure:

10 min

Observation period (in vivo):

2 h

Number of animals or in vitro replicates:

3 replicates

Details on study design:

PREPARATION OF TEST MATERIAL
The test substance was tested directly, without dilution or preparation of a solution.

NEGATIVE CONTROL
Sodium chloride solution: 0.9% NaCl (CAS No. 7647-14-5), dissolved in demineralised water.

POSITIVE CONTROL
Dimethylformamide, DMF, CAS no. 68-12-2, undiluted.

PREPARATIONS
After having carefully cleaned and sterilized the cornea holders, they were kept in the incubation chamber at 32±1°C. On the day of the assay, the MEM without phenol red was supplemented with sodium bicarbonate, L-glutamine and 1% fetal calf serum (complete MEM) and stored in a water bath at 32±1°C. The same was performed with the MEM with phenol red but without addition of sodium bicarbonate. After the arrival of the corneas, they were examined and only corneas which were free from damages were used. The corneas were excised with a scalpel and cut from the globe with a 2 - 3 mm ring of sclera around the outside. Each cornea was transferred to a cornea holder in which pre-warmed cMEM without phenol red was filled. The holders were then incubated for 1 h in the incubation chamber at 32±1°C.

METHOD
After the initial incubation of 1 h, the medium was changed and the baseline opacity for each cornea was recorded at 570 nm. For each treatment group (negative control solution, test substance and positive control), three replicates were used. After removal of the pre-incubation medium, 750 μL were applied to each replicate by closed chamber-method. An exposure period of 10 min was given. After thorough rinsing with cMEM with phenol red and final rinsing with cMEM without phenol red, corneas were stored for additional 2 h at 32±1°C. Then, the final opacity value and permeability of each cornea was recorded.

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Remarks:

mean IVIS

Value:

0.62

Remarks on result:

no indication of irritation

In vivo

Irritant / corrosive response data:

The test substance showed no effects on the cornea of the bovine eye. The calculated IVIS (in vitro irritancy score) was 0.62 under the conditions of this test.

Any other information on results incl. tables

The calculated IVIS for each replicate and the corresponding means are presented in the following table:

Table 1: Calculated and mean IVIS

Test group	IVIS	Mean IVIS	Relative standard deviation IVIS
Negative Control 0.9% NaCl	-0.88	-0.75	-14.8%
	-0.70
	-0.67
Test substance	1.08	0.62	91.7%
	0.80
	-0.02
Positive control DMF undiluted	94.18	105.62	35.3%
	75.43
	147.25

 

The test was considered as valid if the positive control causes an IVIS that falls within two standard deviations of the current historical mean. The negative control has to show an IVIS ≤3. Values for negative and positive controls were within the range of historical data of the test facility. Therefore, the test system was acceptable.

Applicant's summary and conclusion

Interpretation of results:

not irritating

Conclusions:

The test substance is not irritating to eyes.

Executive summary:

An in vitro study was conducted to assess the corneal damage potential of the test substance by quantitative measurements of changes in opacity and permeability in a bovine cornea, according to OECD Guideline 437 and EU Method B.47, in compliance with GLP. The test substance was brought onto the cornea of a bovine eye which previously had been incubated with cMEM without phenol red at 32±1°C for 1 h and whose opacity had been determined. The test substance was incubated on the cornea for 10 min at 32±1°C. After removal of the test substance and 2 h post incubation, opacity and permeability values were measured and compared with the initial value. The negative and positive controls met the validity criteria. The test substance showed no effects on the cornea of the bovine eye. The calculated IVIS (in vitro irritancy score) was 0.62. Under the test conditions, the test substance was found to be not irritating to eyes (Andres, 2015).