Amides, from branched and linear C16-18 and C18-unsatd. fatty acids, diethylenetriamine and tall-oil fatty acids, citrates

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

15-05-2012 to 17-05-2012

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: not formally validated study method

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Principles of method if other than guideline:

The SkinEthic reconstructed human corneal epithelium assay is used

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Fatty acids, tall-oil, reaction products with fatty acids C16-C18 and C18-unsatd, branched and linear, diethylenetriamine and citric acid

EC Number:

700-795-0

Cas Number:

1393571-43-1

Molecular formula:

NA

IUPAC Name:

Fatty acids, tall-oil, reaction products with fatty acids C16-C18 and C18-unsatd, branched and linear, diethylenetriamine and citric acid

Test material form:

other: extremely viscous liquid

Details on test material:

- Name of test material (as cited in study report): Fatty acids, tall oil, reaction products with fatty acids C16-18 and C18-unsatd, branched and linear, diethylenetriamine and citric acid
- Substance type: UVCB
- Physical state: extremely viscous liquid
- Storage condition of test material: room temperature in dark

Test system

Details on study design:

The SkinEthic reconstructed human corneal epithelium assay uses corneal epithelial tissue to assess penetration and concomittant cytotoxicity of the test substance.

In a pre-test direct interaction of the test substance with the detection chemical MTT is assessed.
In the main study triplicate SkinEthic tissues were treated with 30 uL of the test substance during 10 minutes. In addition a negative control (Na2HPO4 0.142 g/L; glucose 1.802 g/L; HEPES 7,149 g/L; KCl 0.224 g/L; NaCl 7.597 g/L) and a positive control (2% SDS) were included. After treatment tissues (2 per treatment) were rinsed and treated with MTT. Optical density of extracted MTT solutions was measured at 540 nm. Viability was expressed as percentage MTT conversion versus negative control.
Tissue viability ≥ 60% means non-irritant
Tissue viability < 60% means irritant
The third tissue per treatment was preserved for histopathology.

Results and discussion

In vitro

Results

Irritation parameter:

other: % tissue viability

Value:

79

Negative controls validity:

valid

Positive controls validity:

valid

Remarks:

25.5% tissue viability

In vivo

Other effects:

The test substance did not interfere with the reduction of MTT to blue formazan salt.
In the main test after rinsing the test substance was found to remain on the tissue surface due to the extreme viscosity.

Any other information on results incl. tables

	Mean OD540	Relative mean viability
Negative control:	0.729	100%
Test substance:	0.626	79%
SDS:	0.202	25.5%

No histopathology was performed.

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: other: < 60% viability is indicative for positive result

Conclusions:

The test substance is non-irritant in the SkinEthic reconstructed human corneal epithelium assay.

Executive summary:

In this assay triplicate SkinEthic tissues were treated with 30 uL of the test substance during 10 minutes. After treatment tissues (2 per treatment) were rinsed and treated with MTT. Optical density of extracted MTT solutions was measured at 540 nm. Viability was expressed as percentage MTT conversion versus negative control. The viability was 79%, which is indicative for non-irritant substances.

As the assay is not formally validated, the results need to be regarded as preliminary.