Disodium sebacate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2016-06-22 to 2016-08-10

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

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:

26 July 2013

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

BASF SE, Experimental Toxicology and Ecology, 67056 Ludwigshafen, Germany

Test material

Test material information

Constituent 1

Reference substance name:

Disodium sebacate

EC Number:

241-300-3

EC Name:

Disodium sebacate

Cas Number:

17265-14-4

Molecular formula:

C10H18O4.2Na

IUPAC Name:

disodium sebacate

Test material form:

solid

Specific details on test material used for the study:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test-substance preparation was solved in H2O per weight per volume (w/v) and stirred with a magnetic stirrer shortly before application.

Test animals / tissue source

Species:

cattle

Strain:

not specified

Details on test animals or tissues and environmental conditions:

SOURCE OF COLLECTED EYES
- Source: Schlachthof Mannheim, Schlachthofstr. 21, 68165 Mannheim, Germany
- Characteristics of donor animals: age min. 12 months, max. 60 months
- indication of any existing defects or lesions in ocular tissue samples: Only Corneas free of defects were used

Test system

Vehicle:

water

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Concentration (if solution):
Treatment: 750 µL, 20 % (w/v) test substance in water
Negative control: 750 µL de-ionized water
Positive control: 750 µL, 20 % (w/v) imidazole/de-ionized water

Duration of treatment / exposure:

Corneal opacity: 4 h at 32 °C

Duration of post- treatment incubation (in vitro):

Permeability: replacement of the test substance with 1 mL sodium fluorescin solution (5 mg/mL) and 90 ±5 min. incubation at 32 °C

Number of animals or in vitro replicates:

3 corneas in each group (Treatment, NC, PC)

Details on study design:

SELECTION AND PREPARATION OF CORNEAS
Corneas free of defects (opacity, scratches, pigmentation etc.) were dissected with a 2 to 3 mm rim of sclera. Isolated corneas were mounted in cornea holders that consists of anterior and posterior chambers. Both chambers were filled to excess with pre-warmed Eagle’s MEM (without phenol red) and then equilibrated in a vertical position at about 32 °C for at least 1 hour.

QUALITY CHECK OF THE ISOLATED CORNEAS
After the equilibration period the medium in both chambers was replaced with fresh prewarmed medium and initial corneal opacity readings were taken for each cornea with an opacitometer. Any corneas that showed macroscopic tissue damage or an opacity value < 548 opacity units were discarded.

NUMBER OF REPLICATES
3

NEGATIVE CONTROL USED
Yes, 750 µL de-ionized water

POSITIVE CONTROL USED
Yes, 20% (w/v) solution of imidazole in de-ionized water

POST-INCUBATION PERIOD
no

REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period: at least 3 times with Eagle’s MEM (containing phenol red) and once with Eagle’s MEM (without phenol red).

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: corneal opacity readings were taken for each cornea with an opacitometer
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of a 96-well microtiter plate reader (OD490)

SCORING SYSTEM:
In Vitro Irritancy Score (IVIS)
Evaluation of the test results based on to the criteria given in OECD 437 (July 2013).

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Run / experiment:

BCOP Test

Value:

0.1

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

OTHER EFFECTS:
- Visible damage on test system: Not reported

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Yes
- Acceptance criteria met for positive control: Yes

Any other information on results incl. tables

BCOP Test results:

Test substance identification	Mean Opacity Value	Mean Permeability Value	Mean In Vitro Irritancy Score
Test substance	0.0	0.004	0.1
NC	10.6	0.003	10.6
PC	84.5	2.259	118.4

Applicant's summary and conclusion

Interpretation of results:

other: not severely irritating

Conclusions:

Based on the results for BCOP and EpiOcular Test and applying the evaluation criteria described, the test item shows an eye irritation potential in the in vitro eye irritation test strategy under the test conditions chosen but is not severely irritating or corrosive.

Executive summary:

The potential of the test tem to cause ocular irritation or serious damage to the eyes was assessed by a single topical application of 750 μL of a 20% test-substance preparation to the epithelial surface of isolated bovine corneas. Three corneas were treated with the test-substance preparation for an exposure period of 4 hours. In addition to the test substance a negative control (NC; de-ionized water) and a positive control

(PC; 20% imidazole in de-ionized water) were applied to three corneas each. Corneal opacity was measured quantitatively as the amount of light transmitted through the cornea. Permeability was measured quantitatively as the amount of sodium fluorescein dye that passes across the full thickness of the cornea. Both measurements were used to calculate an In Vitro Irritancy Score of the test substance.

Based on the results for BCOP and EpiOcular Test and applying the evaluation criteria described, the test item shows an eye irritation potential in the in vitro eye irritation test strategy under the test conditions chosen but is not severely irritating or corrosive.