Fatty acids, tall-oil, esters with ethylene glycol

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

30 Apr - 04 May 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 492 (Reconstructed Human Cornea-like Epithelium (RhCE) Test Method for Identifying Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage)

Version / remarks:

adopted Jun 2018

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Fatty acids, tall-oil, esters with ethylene glycol

EC Number:

269-130-5

EC Name:

Fatty acids, tall-oil, esters with ethylene glycol

Cas Number:

68187-85-9

Molecular formula:

C38H66O4 C38H64O4 C40H68O4 C40H70O4 C38H68O4

IUPAC Name:

2-[(9Z)-octadec-9-enoyloxy]ethyl (9Z)-octadec-9-enoate; 2-[(9Z)-octadec-9-enoyloxy]ethyl (9Z,12Z)-octadeca-9,12-dienoate; 2-[(9Z,12Z)-octadeca-9,12-dienoyloxy]ethyl (9Z,12Z)-octadeca-9,12-dienoate

Test animals / tissue source

Species:

human

Strain:

other: Reconstructed Human EpiOcular™ Model

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 50 µL

NEGATIVE CONTROL
- Amount(s) applied: 50 µL

POSITIVE CONTROL
- Amount(s) applied: 50 µL

Duration of treatment / exposure:

30 ± 2 min at 37.0 ± 1.0°C

Duration of post- treatment incubation (in vitro):

12 ± 2 min at room temperature (post-exposure immersion) and 120 ± 15 min at 37 °C (post-incubation)

Number of animals or in vitro replicates:

Duplicate measurements each of 2 independent tissues for treatment, negative and positive control group, respectively

Details on study design:

- RhCE tissue construct used, including batch number : EpiOcular™ (OCL-200-EIT MatTek Corporation, Lot: 27427 Kit A)

- Indication of controls used for direct MTT-reducers and/or colouring test chemicals: The ability of the test substance to directly reduce MTT and to change the color of the MTT medium was assessed in a pre-experiment. Because no color changes were observed it was concluded that the test item did not interact with the MTT endpoint and did not induce color interference.

- Description of the method used to quantify MTT formazan : The amount of extracted formazan was determined spectrophotometrically at 570 nm in duplicate with the TECAN Infinite® M200 Pro Plate Reader. Cell viability was calculated for each tissue as a percentage of the mean of the negative control tissues. Eye hazard potential of the test item was classified according to remaining cell viability following exposure of the test item.

- Description of evaluation criteria used: The test substance is considered to be not irritating to eye if the tissue viability is > 60%. The test substance is considered to be irritating to eye if the tissue viability is ≤ 60%.

- Positive and negative control means and acceptance ranges: OD in the negative control substance group is > 0.8 and < 2.5. The cell viability in the positive control substance group is < 50%.
- Acceptable variability between tissue replicates for positive and negative controls and for the test substance: Differences of two tissue cell viabilities in each treatment group are < 20%

Results and discussion

In vitro

Results

Irritation parameter:

other: % tissue viability mean value of 2 tissues

Run / experiment:

30 ± 2 min

Value:

102

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

OTHER EFFECTS:
- Visible damage on test system:

DEMONSTRATION OF TECHNICAL PROFICIENCY:

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Yes, the OD of the negative control was 1.9660, 1.9044, 1.6966 and 1.6817, and thus within the acceptibility range (OD > 0.8 and < 2.5).
- Acceptance criteria met for positive control: Yes, the positive control reduced the cell viability at 2.9% and fulfilled the acceptance criteria.
- Acceptance criteria for variabilties: Differences of two tissue cell viabilities in the negative control substance, the positive control substance and the test substance groups were 14%, 4.2% and 1.3%%, respectively, and thus < 20%.

Any other information on results incl. tables

Table 1: Individual absorption values

	Tissue A	Tissue B
	(OD570)	(OD570)
Negative control measurement 1	19.660	16.966
Negative control measurement 2	19.044	16.817
Test item measurement 1	19.030	18.244
Test item measurement 2	18.664	17.970
Positive control measurement 1	0.1035	0.0807
Positive control measurement 2	0.1027	0.0801

OD: Optical density

Table 2: Mean tissue viability

	Mean tissue viability (% of control)	Difference between two tissues  (%)
Negative control	100	14
Test item	102	4.2
Positive control	2.9	1.3

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