Fatty acids, sunflower-oil, conjugated, maleated,reation products with diethanolamine, maleated tall-oil fatty acids and triethanolamine

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

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:

2017

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 i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)

Version / remarks:

July 2013

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:

Dec. 2010

Test guideline 3

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:

July 2015

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Fatty acids, sunflower-oil, conjugated, maleated,reation products with diethanolamine, maleated tall-oil fatty acids and triethanolamine

EC Number:

800-003-4

Cas Number:

1415316-96-9

IUPAC Name:

Fatty acids, sunflower-oil, conjugated, maleated,reation products with diethanolamine, maleated tall-oil fatty acids and triethanolamine

Test material form:

liquid: viscous

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 1089651-lab
- Expiration date of the lot/batch: March 14, 2020
- Purity test date: May 24th, 2017

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature
- Stability under test conditions: guaranteed by sponsor

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: For better handling, the test substance was heated for about an hour at 37°C. Before application, it was cooled down to room temperature.
- Preliminary purification step (if any): Removal of water content to get a pure, i.e., solvent free sample of the registered substance for testing (Residual water content in the final sample was app. 2%)

Test animals / tissue source

Species:

other: Isolated bovine cornea (BCOP) and human-derived, epidermal keratinocytes (EpiOcular)

Details on test animals or tissues and environmental conditions:

BCOP: SOURCE OF COLLECTED EYES
- Source: Schlachthof Mannheim, Schlachthofstr. 21, 68165 Mannheim, Germany
- Characteristics of donor animals: Age: 12 - 60 months
- indication of any existing defects or lesions in ocular tissue samples: Corneas were free of defects (scratches, opacity, pigmentation, etc.)

Epioclular
The EpiOcularTM model (OCL-200) is a three-dimensional, non-keratinized tissue construct composed of normal human-derived, epidermal keratinocytes used to model the human corneal epithelium (compare Figure 1). The EpiOcularTM tissues (surface 0.6 cm²) are cultured on cell culture inserts (MILLICELLs, 10 mm diameter) and are commercially available as kits (EpiOcular™ 200) containing 24 tissues on shipping agarose.

Tissue Model: OCL-200
Lot number: 23789 (test run 1), 23797 (test run 2)
Supplier: MatTek In Vitro Life Science Laboratories, Bratislava, Slovakia
The supplier demonstrated intakt barrier function (ET50 values between 12.2 and 37.5min)

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

The test substance was heated for 1 hour prior to application to 37°C
- Amount(s) applied: 750µL (BCOP), 50µL (EpiOcular)

Duration of treatment / exposure:

10 minutes (BCOP)
30min (EpiOcular)

Duration of post- treatment incubation (in vitro):

2h (BCOP and EpiOcular)

Number of animals or in vitro replicates:

3 (BCOP), 2 (EpiOcular)

Details on study design:

BCOP
METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: Before measurement, each cornea was visually observed and observations were recorded. Final corneal opacity readings were taken for each cornea with an opacitometer.
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of a microtiter plate reader (OD490)

SCORING SYSTEM: In Vitro Irritancy Score (IVIS)

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

cornea opacity score

Run / experiment:

BCOP

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

fluorescein retention score

Run / experiment:

BCOP

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

in vitro irritation score

Run / experiment:

BCOP

Value:

0

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Results 4

Irritation parameter:

other: viability

Run / experiment:

EpiOcular, 1st run

Value:

62.3

Negative controls validity:

valid

Positive controls validity:

valid

Results 5

Irritation parameter:

other: viability

Run / experiment:

EpiOcular, 2nd run

Value:

95.4

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