Reaction products of fatty acids, tall oil and fatty acids, C18 unsaturated, trimers and fatty acids, C18 unsaturated, dimers with (9Z)-octadec-9-en-1-amine

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study meeting generally accepted scientific standards and documented in detail

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Qualifier:

no guideline followed

Principles of method if other than guideline:

The present test is based on the experience that irritant chemicals produce cytotoxicity in human reconstructed cornea after a short term topical exposure. After application of 50µl of the test material to the surface of the three dimensional human cornea model "EpiOcular" for 30minutes, the test substance is removed. After a 2h post-exposure period, the induced cytotoxicity (= loss of viability) is measured as the reduction of mitochondrial dehydrogenase activity using MTT. A chemical is considered as irritant, if the mean viability of smaller or equal 50%, and as not-irritant, if the mean viability is above 60%. In the range between 50% and 60% no prediction can be made.

Though there are no official national or international guidelines for the EpiOcularTM test yet, the study was performed according to the methods described in the following publications:
- MatTek Corporation, Ashland, MA 01721, USA: EpiOcularTM human cell construct: Procedure details, Version 3.1a of February 10, 2010.
- Harbell J.W. et al. (2009): COLIPA Program on Optimization of Existing In Vitro Eye Irritation Assays for Entry into Formal Validation: Technology Transfer and Intra/Inter Laboratory Evaluation of EpiOcular Assay for Chemicals. Poster # 378, Society of Toxicology, March 2009.
In addition the study follows the testing strategy for determination of eye irritation/corrosion as given in the following OECD guideline:
- OECD Guideline for Testing of Chemicals No. 405, April 24, 2002 (“Acute Eye Irritation/Corrosion”)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Reaction products of Fatty acids, tall-oil, compds. with oleylamine and Fatty acids, C18-unsatd., trimers, compds. with oleylamine

EC Number:

942-330-6

Molecular formula:

Not applicable (UVCB substance)

IUPAC Name:

Reaction products of Fatty acids, tall-oil, compds. with oleylamine and Fatty acids, C18-unsatd., trimers, compds. with oleylamine

Details on test material:

- Name of test material (as cited in study report): Reaction mass of Fatty acids, tall-oil, compds. with oleylamine and Fatty acids, C18-unsatd., trimers, compds. with oleylamine
- Physical state: liquid
- Analytical purity:
- Purity test date:
- Lot/batch No.: D987-AM-202011
- Expiration date of the lot/batch:
- Storage condition of test material:

Test animals / tissue source

Species:

other: human cornea model

Details on test animals or tissues and environmental conditions:

Three dimensional human cornea model EpiOcular OCL-200
- Source: MatTek corp., Ashland MA, USA
- Culture Medium: DMEM
- Cell death detection agent: 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) (MatTek corp., Ashland MA, USA / Sigma,Germany)
- duplicate cultures
Since the test substance is able to reduce MTT directly, a freeze killed tissue sample each was treated with the test substance or the negative control in the same way as all other tissue samples. However, a difference below 0.1 in MTT reduction between test sample and negative control was obtained in this experiment. Thus, the results were not used for correction of the OD750 values obtained in the viable cultures.

Test system

Vehicle:

unchanged (no vehicle)

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 50µl

Duration of treatment / exposure:

30min

Details on study design:

Negative control: De-ionized water
Positive control: Methyl acetate (98%+)

Incubation times:
Pre-incubation in culture medium: 16-24h
Pre-treatment with PBS to wet the tissue surfact for 30min
Exposure time: 30min
Post-incubation period: 2h (after washing)
MTT incubation: 3h

REMOVAL OF TEST SUBSTANCE
- Washing (if done): with PBS and medium subsequently at the end of the exposure time. After soaking for 12minutes in medium, the tissues were dried on absorbant paper.

Results and discussion

In vitro

Results

Irritation parameter:

other: cell viability

Run / experiment:

mean (2.5 h)

Value:

83

Remarks on result:

other: not irritating

Remarks:

value: 83 %

Applicant's summary and conclusion

Interpretation of results:

not irritating