Fatty acids, vegetable-oil, Me esters, sulfurized

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

24 Oct. 2017

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:

2018

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:

Fatty acids, vegetable-oil, Me esters, sulfurized

EC Number:

276-337-4

EC Name:

Fatty acids, vegetable-oil, Me esters, sulfurized

Cas Number:

72102-30-8

IUPAC Name:

Fatty Acids, vegetable-oil, Me-Esters, sulfurized

Test material form:

liquid

Details on test material:

- Density: 0.95 g/mL
- Batch no.: 71012767

Test animals / tissue source

Species:

cattle

Strain:

other: Bos primigenius Taurus

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

750µL

Duration of treatment / exposure:

10 minutes

Duration of post- treatment incubation (in vitro):

2 hours

Number of animals or in vitro replicates:

3

Details on study design:

SELECTION AND PREPARATION OF CORNEAS
The corneas 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 (32 ± 1 °C) without phenol red was filled. The holders were then incubated for 1 hour in the incubation chamber at 32 ± 1 °C.

QUALITY CHECK OF THE ISOLATED CORNEAS
After the initial incubation, the medium was changed and the baseline opacity for each cornea was recorded. None of the corneas showed tissue damage; therefore, all corneas were used.

NUMBER OF REPLICATES
For each treatment group (negative control solution, test item and positive control), three replicates were used.
NEGATIVE CONTROL USED
HBSS: Hank’s Balanced Salt Solution (HBSS) 10-fold concentrated, diluted in demin. water (1:10), batch no.: 20171024

POSITIVE CONTROL USED
Dimethylformamide, DMF, CAS-No. 68-12-2, undiluted, batch no.: 475235719

APPLICATION DOSE AND EXPOSURE TIME
750 μL negative control solution, test item and positive control were applied to each replicate.

TREATMENT METHOD:
closed chamber

POST-INCUBATION PERIOD:
yes; 2 hours

REMOVAL OF TEST SUBSTANCE
After thorough rinsing with cMEM with phenol red and final rinsing with cMEM without phenol red, the anterior chamber was filled with cMEM without phenol red. After post-incubation time, the cMEM without phenol red was renewed in both chambers. Then, the final opacity value of each cornea was recorded.

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity:
The change of opacity value of each treated cornea with test item, positive control and negative control was calculated by subtracting the initial basal opacity from the post treatment opacity reading for each cornea. The average change in opacity of the negative control cornea was calculated and this value was subtracted from the change in opacity of each treated cornea with test item and positive control to obtain a corrected opacity.

- Corneal permeability:
Passage of sodium fluorescein dye measured with the aid of microtiter plate photometer (OD492)

SCORING SYSTEM:
In Vitro Irritancy Score (IVIS)

DECISION CRITERIA:
According to OECD Guideline no. 437 (Jul. 2013), a substance with an IVIS ≤ 3 requires no classification for eye irritation or serious eye damage.

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Run / experiment:

For each treatment group (negative control solution, test item and positive control), three replicates were used. Duration of each exposure: 10 minutes

Value:

0.79

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of this study, the test item Fatty Acids, vegetable-oil, Me-Esters,
sulfurized showed no effects on the cornea of the bovine eye. The calculated IVIS is 0.79.
According to OECD Guideline no. 437 (Jul. 2013), a substance with an IVIS ≤ 3 requires
no classification for eye irritation or serious eye damage.