Barium(2+) 12-hydroxyoctadecanoate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

from 2018-01-09 to 2018-01-18

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

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:

09 October 2017

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Barium(2+) 12-hydroxyoctadecanoate

EC Number:

244-469-1

EC Name:

Barium(2+) 12-hydroxyoctadecanoate

Cas Number:

21598-22-1

Molecular formula:

C18H36O3.1/2Ba

IUPAC Name:

barium bis(12-hydroxyoctadecanoate)

Test material form:

solid

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Batch No.of test material: Lab NP_20171034-003
- Expiration date of the lot/batch: 2022-08-18

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: closed vessel at room temperature (20±5°C).

OTHER SPECIFICS:
white solid

Test animals / tissue source

Species:

human

Details on test animals or tissues and environmental conditions:

- Justification of the test method and considerations regarding applicability: The test system was chose according to the OECD guideline 492.
- Description of the cell system used, incl. certificate of authenticity and the mycoplasma status of the cell live: The EpiOcularTM tissue consists of normal, human-derived keratinocytes which have been cultured to form a stratified squamous epithelium similar to that found in the human cor¬nea. It consists of highly organized basal cells. These cells are not transformed or trans-fected with genes to induce an extended life span. The EpiOcularTM tissues are cultured in specially prepared cell culture inserts with a porous membrane through which nutrients can pass to the cells. The tissue surface is 0.6 cm². For further information see "Any other information on materials and methods".

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount applied: ca. 50 mg

Duration of treatment / exposure:

6 h

Duration of post- treatment incubation (in vitro):

18 h

Number of animals or in vitro replicates:

2

Details on study design:

- Details of the test procedure used
- RhCE tissue construct used, including batch number: EpiOcular™ tissues were procured from MatTek In Vitro Life Science Laboratories, Mylnske Nivy 73, 82105 Bratislava, Slovakia; Batch No: 27019
- Doses of test chemical and control substances used
Test item: ca. 50 mg
positive and negative control: 50 µL each
- Duration and temperature of exposure, post-exposure immersion and post-exposure incubation periods:
exposure: 6 h
post treatment: 18 h
- Number of tissue replicates used per test chemical and controls (positive control, negative control): 2
- Wavelength used for quantifying MTT formazan: 570 nm
- Description of the method used to quantify MTT formazan: MTT assay via 96-well plate reader

Results and discussion

In vitro

Results

Irritation parameter:

other: % cell viability

Value:

108.9

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

OTHER EFFECTS:
- Visible damage on test system: No

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

Any other information on results incl. tables

Table 5: Validity of the Experiment

Criterion	Demanded	Found
OD of negative control	> 0.8 and < 2.5	1.7
% mean relative viability of positive control	< 50% of negative control	31.5%
Variation within replicates	< 20%	3.8% (negative control) 6.4% (positive control) 3.4% (test item)

Table 6: Absorbance Values Blank Isopropanol (OD at 570 nm)

Replicate	1	2	3	4	5	6	7	8	Mean
Absorbance	0.036	0.038	0.037	0.038	0.037	0.038	0.038	0.039	0.038

Table 7: Absorbance Values Negative Control, Positive Control and Test Item (OD at 570 nm)

Designation	Measurement	Negative Control	Positive Control	Test Item
Tissue 1	1	1.755	0.603	1.779
	2	1.738	0.638	1.893
Tissue 2	1	1.669	0.500	1.886
	2	1.697	0.525	1.900

Table 8: Mean Absorbance Negative Control, Positive Control and Test Item

Designation	Negative Control	Positive Control	Test item
Mean - blank (Tissue 1)	1.709	0.583	1.798
Mean - blank (Tissue 2)	1.645	0.475	1.855

Table 9: % Viability Positive Control and Test Item

Designation	Positive Control	Test Item
% Viability (Tissue 1)	34.7%	107.2%
% Viability (Tissue 2)	28.3%	110.6%
% Viability Mean	31.5%	108.9%
standard deviation	4.5%	2.4%

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

In the in vitro eye irritation assay (RhCE) according to OECD guideline 492, a mean tissue viability of 108.9% was determined.

Executive summary:

The eye irritating potential of the test item was determined in an in vitro eye irritation assay (RhCE) according to OECD guideline 492. The test item was applied to a three-dimensional human cornea tissue model in duplicate for an exposure time of 6 hours. The solid test item was applied to two tissue replicates. After treatment, the respective substance was rinsed from the tissue and a post treatment incubation of 18 h followed. Cell viability of the tissues was evaluated by addition of MTT, which can be reduced to formazan. The formazan production was evaluated by measuring the optical density (OD) of the resulting solution. Demineralised water was used as negative control and methyl acetate was used as positive control. After treatment with the negative control, the absorbance values were within the required acceptability criterion of mean OD > 0.8 and < 2.5, OD was 1.7. The positive control showed clear eye irritating effects, mean value of the relative tissue viability was 31.5 % (< 50%). Variation within tissue replicates was acceptable (< 20%). After treatment with the test item, the mean value of relative tissue viability was 108.9 %. This value is above the threshold for eye irritation potential (< 60%).

Under the conditions of the test, the test item is considered non-eye irritant in the EpiOcular Eye Irritation Test.