Tall-oil fatty acids salts of condensation products of tall-oil fatty acids with diethanolamine and triethanolamine

Eye irritation

Administrative data

Endpoint:

eye irritation

Remarks:

other: in vitro

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Well documented and reported study fully adequate for assessment. The study was conducted according to an internationally accepted technical guideline and in compliance with GLP in a recognized contract research organization.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: OECD Guideline 437 of July 2013: 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

Test material form:

liquid: viscous

Test animals / tissue source

Species:

other: in vitro test

Strain:

other: bovine eyes

Test system

Vehicle:

physiological saline

Amount / concentration applied:

750 µl of test substance, positive control solution (ethanol) or negative control solution (physiol. saline) was introduced into the anterior part of each cornea holder. Following application, turning of the holder into horizontal position and its slight rotation ensured uniform distribution of the test substance over the surface of the cornea.

Duration of treatment / exposure:

10 min (± 30 sec.).
Each cornea holder was incubated in a horizontal position at 32°C ± 1°C in a waterbath. Following incubation, the test substance, positive and negative controls were removed and the epithelial surface of the cornea washed, at least three times or until the wash medium (EMEM* with phenol red) was clear and there was no discolouration.

* EMEM, Eagles Minimal Essential Medium

Details on study design:

Eyes were maintained and transported to the laboratory and isolated corneas stored in sufficient HBSS* containing 1% penicillin-streptomycin solution (10’000 IU/ml and 10 mg/ml, respectively) until being used. The eyes were used within 4 hours of slaughter. Each treatment group (test substance, negative/positive controls) comprised 3 corneas from bovine eyes. Each cornea surrounded by ca. 2 to 3 mm of sclera was mounted to a holder. Adequate exposure of the epithelial surface of the cornea to the test substance was provided. Opacity of the cornea was determined by measurement of light transmission through the centre of each mounted cornea using a calibrated opacitometer. Permeability of the cornea was determined by spectrophotometric measurement at 490 nm (OD490) of medium from the posterior compartment of the cornea holder after addition of 1 ml of sodium fluorescein solution (4 mg/ml) to the anterior compartment, thus determining the degree of penetration of sodium fluorescein through all corneal cell layers. Prior to the spectrophotometric measurement, incubation with fluorescein solution had happened in horizontal position of the cornea holder at 32°C ± 1°C for 90 ± 5 minutes in a water bath. Then the medium in the posterior compartment was mixed by drawing approximately 2.5 ml gently up and down a 5 ml syringe, with a needle attached, three times. Any solution giving an OD490 value above 1.8 was diluted 1 in 5 with cMEM.

Effects to the cornea (corresponding to possible eye damage) were measured as opacity and permeability, which when combined gave an In Vitro Irritancy Score (IVIS) for each treatment group. A substance that induces an IVIS greater than 55 is defined as a corrosive or severe irritant.

*HBSS Hanks Balanced Salt Solution containing Ca++ and Mg++ without phenol red

Results and discussion

In vivo

Results

Irritation parameter:

other: in vitro irritancy score (IVIS)

Basis:

mean

Remarks:

calculated from the IVIS of each of 3 isolated bovine corneas

Score:

0.5

Remarks on result:

other: IVIS = Corrected Opacity Value + (15 x Corrected permeability OD490 Value)

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information