Octanoic acid, 2-phenoxyethyl ester

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

11 Dec 2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

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)

Version / remarks:

adopted in 2013

Deviations:

yes

Remarks:

the study was conducted prior to the adoption of the test guideline, according to the draft version (dated 14 Aug 2008)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

2-phenoxyethyl octanoate

EC Number:

607-240-0

Cas Number:

23511-73-1

Molecular formula:

C16H24O3

IUPAC Name:

2-phenoxyethyl octanoate

Test animals / tissue source

Species:

cattle

Strain:

not specified

Details on test animals or tissues and environmental conditions:

SOURCE OF COLLECTED EYES
- Source: Slaughterhouse (Vitelco, 's-Hertogenbosch, the Netherlands)
- Storage, temperature and transport conditions of ocular tissue: Eyes were collected and transported in physiological saline in a suitable container

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 750 µL

CONTROLS
- Negative control: 750 µL
- Positive controls: 750 µL
- Concentration of positive control solution: Alkylbenzyldimethylammonium chloride 10% (w/v) in physiological saline and 99.9% ethanol

Duration of treatment / exposure:

10 ± 1 min

Duration of post- treatment incubation (in vitro):

120 ± 10 min

Number of animals or in vitro replicates:

3

Details on study design:

SELECTION AND PREPARATION OF CORNEAS
Corneas were isolated and cultured in cMEM supplemented with 1% (v/v) foetal bovine serum and 1% (v/v) L-glutamine at 32 ± 1 °C for a minimum of 1 h prior to the start of the experiment. Opacity determinations were performed on each of the corneas using an opacitometer. Corneas that had an initial opacity reading higher than 3 were not used. Three corneas were selected at random for each treatment group.

QUALITY CHECK OF THE ISOLATED CORNEAS
Eyes were carefully examined for defects by holding the eyes submersed in physiological saline; those exhibiting unaccetable defects (opacity, scratches, pigmentation, neovascularisation) were discarded

TREATMENT METHOD
Each cornea was mounted in a cornea holder with the endothelial side against the O-ring of the posterior part of the holder. Afterwards, the anterior part of the holder was positioned on top of the cornea and fixed in place with screws. Both compartments of the holder were filled with incubation medium. For equilibration, the corneas in the holder were incubated in at 32 ± 1 °C for at least one hour.

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

METHODS FOR MEASURED ENDPOINTS
- Corneal opacity: Corneal opacity was determined by the amount of light transmission through the cornea via an opacitometer.
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of microtiter plate reader (OD490)

SCORING SYSTEM
In Vitro Irritancy Score (IVIS)
In vitro score range In vitro classification
0 - 3 Non irritant
3.1 - 25 Mild irritant
25.1 - 55 Moderate irritant
55.1 - 80 Severe irritant
> 80 Very severe irritant

DECISION CRITERIA
The mean opacity and mean permeability values (OD490) were used for each treatment group to calculate an in vitro score:
In vitro irritancy score (IVIS) = mean opacity value + (15 x mean OD490 value)

The positive control in vitro irritancy score should be reasonably within the laboratory historical positive control data range.
- The uncorrected negative control in vitro irritancy score is less than 3.1

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

in vitro irritation score

Run / experiment:

mean out of 3 eyes

Value:

0.1

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks:

IVIS score of 157.9 for Alkylbenzyldimethylammonium chloride and 68.3 for ethanol, respectively

Results 2

Irritation parameter:

other: Opacity

Run / experiment:

mean out of 3 eyes

Value:

0

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks:

Mean opacity of 86 for Alkylbenzyldimethylammonium chloride and 24 for ethanol, respectively

Results 3

Irritation parameter:

other: Permeability

Run / experiment:

mean out of 3 eyes

Value:

0.009

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks:

Mean permeability of 4792 for Alkylbenzyldimethylammonium chloride and 2950 for ethanol, respectively

Other effects / acceptance of results:

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: The individual in vitro irritancy scores for the negative controls was 0 for all three corneas.
- Acceptance criteria met for positive control: The individual positive control in vitro irritancy scores ranged from 145 to 165 for benzalkonium chloride and ranged from 60 to 83 for ethanol and were within the range of historical control data. The corneas treated with the positive control substances were opaque after the 10 minutes of treatment. Thus, the test system functioned properly.
- Range of historical values if different from the ones specified in the test guideline:
- Negative control: Range = -1.0 - 2.2; Mean = 0.3; SD = 0.6; n = 41
- Positive control: Range = 113.8 - 202.7; Mean = 163.4; SD = 22.1; n = 42

Any other information on results incl. tables

Table 1: Individual results

	Eye No.	corrected final opacity	corrected final OD 450	in vitro irritancy score
				individual	mean
negative control	1	0	0.003	0	0
	2	0	0	0
	18	0	-0.002	0
positive control A	4	88	3.78	144.7	157.9
	5	89	5.07	165.1
	6	81	5.526	163.9
positive control B	16	26	3.816	83.2	68.3
	17	23	2.49	60.4
	3	23	2.544	61.2
test substance	13	0	0.006	0.1	0.1
	14	0	0.023	0.3
	15	0	-0.002	0

A Benzalkonium chloride

B Ethanol

Applicant's summary and conclusion

Interpretation of results:

other: CLP/EU GHS criteria not met, no classification required according to Regulation(EC) No. 1272/2008