Reaction mass of Fatty acids, tallow, Guerbet reaction Products, Ca salts and Fatty acids, tallow, Guerbet reaction Products, Na salts and Fatty acids, tallow, Guerbet reaction Products

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

14 September 2015 - 15 September 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

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

Test material

Test material information

Constituent 1

Reference substance name:

73138-58-6, 73138-59-7, 73138-60-0

IUPAC Name:

73138-58-6, 73138-59-7, 73138-60-0

Details on test material:

- Name of test material (as cited in study report): LICOWAX R 21 S FL
- Physical state: Slightly yellow flakes
- Analytical purity: 99.6 % (w/w)

Fatty acids, tallow,
Guerbet reaction Products 73138-58-6 21% (w/w)
Fatty acids, tallow,
Guerbet reaction Products, Ca salts 73138-59-7 48% (w/w)
Fatty acids, tallow,
Guerbet reaction Products, Na salts 73138-60-0 31% (w/w)

- Lot/batch No.: DEF2084336
- Expiration date of the lot/batch: 2018-11-13
- Storage condition of test material: Room temperature, protected from light in the tightly closed original container
- Solubility : ≤ 0.01 g/L (20 °C)

Test animals / tissue source

Species:

cattle

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Since no workable suspension of the test item could be obtained, it was added pure on top of the corneas.
- Amount applied: 309.8 to 386.0 mg per cornea, ensuring complete coverage

NEGATIVE CONTROL
- Amount applied: 750 µl of physiological saline per cornea

POSITIVE CONTROL
Amount applied: 750 µl 20% (w/v) Imidazole solution

Duration of treatment / exposure:

240 (+/- 10 minutes)

Number of animals or in vitro replicates:

Test item: 3
Negative control: 3
Positive control: 3

Details on study design:

TEST SITE
- Isolated bovine cornea
- Source: Bovine eyes from young cattle were obtained from the slaughterhouse (Vitelco, -'s Hertogenbosch, The Netherlands), where the eyes were excised by a slaughterhouse employee as soon as possible after slaughter. Eyes were collected and transported in physiological saline in a suitable container under cooled conditions.

REMOVAL OF TEST SUBSTANCE
- Washing: yes, at least three times with MEM (with phenol red)
- Time after start of exposure: 240 minutes
- Determination of permeability: After exposure the corneas (anterior compartments) were incubated in sodium-fluorescein (5 mg Na-fluorescein/ml cMEM solution) for 90 ± 5 minutes (both incubations at 32 ± 1°C). The optical density at 490 nm (OD490) was measured in triplicate using a microplate reader.

SCORING SYSTEM:
- The mean opacity and mean permeability values (OD490) were used for each treatment group to calculate an in vitro score:
In vitro irritancy score (IVIS) = mean opacity value + (15 x mean OD490 value).

TOOL USED TO ASSESS SCORE:
- opacitymeter (OP-KIT) and microplate reader (TECAN Infinite® M200 Pro Plate Reader)

DATA EVALUATION:
A test substance that induces an IVIS ≤ 3 is not classified for eye irritancy (UN GHS: no category);
A test substance that induces an IVIS > 55 is defined as a corrosive or severe irritant (UN GHS: catgegory 1);
For a test substance that induces an IVIS >3 and ≥ 55, no prediction on irritant potency can be made.

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Run / experiment:

Mean of 3 replicates

Value:

-1.3

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

- The individual in vitro irritancy scores for the negative controls ranged from -2.7 to -0.4. The individual positive control in vitro irritancy scores ranged from 96 to 149. The corneas treated with the positive control were turbid after the 240 minutes of treatment.
- The corneas treated with test item showed opacity values ranging from -2.1 to -1.2 and permeability values ranging from 0.007 to 0.032. The corneas were clear after the 240 minutes of treatment. No pH effect of the test item was observed on the rinsing medium. Hence, the in vitro irritancy scores ranged from -2.0 to -0.7 after 240 minutes of treatment.
- Negative and positive control responses were within the historical data range, therefore the acceptability criteria were met and the system was considered to function properly.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Remarks:

(also not classified according to CLP criteria)

Conclusions:

Based on the outcome of a Bovine Corneal Opacity and Permeability test (BCOP) performed according to OECD guideline 437 and GLP principles, it is concluded that Licowax R 21 S FL is not irritant or corrosive for the eye under the experimental conditions described in this report. Therefore, no classification is required according to GHS and CLP criteria.

Executive summary:

The eye damage of Licowax R 21 S FL was tested through topical application for approximately 240 minutes.

Since no workable suspension in physiological saline could be obtained, the test item was used as delivered and added pure on top of the corneas (309.8 to 386.0 mg).

The negative control responses for opacity and permeability were less than the upper limits of the laboratory historical range indicating that the negative control did not induce irritancy on the corneas. The mean in vitro irritancy score of the positive control (20% (w/v) Imidazole) was 124 and within two standard deviations of the current historical positive control mean. It was therefore concluded that the test conditions were adequate and that the test system functioned properly.

Licowax R 21 S FL did not induce ocular irritation through both endpoints, resulting in a mean in vitro irritancy score of -1.3 after 240 minutes of treatment.

Since Licowax R 21 S FL induced an IVIS ≤ 3, no classification is required for eye irritation or serious eye damage.