Reaction mass of L-Glutamic acid, N-coco acyl derivs., monosodium salts and sodium hydrogen N-(1-oxooctadecyl)-L-glutamate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

The study was performed between 08 July 2008 and 10 July 2008.

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study conducted in line with good scientific principles and reported to a high standard using a non-validated in vitro method.

Remarks:

Study conducted on read-across material

Justification for type of information:

A RAAF report will shortly be provided.

Cross-reference

Reason / purpose for cross-reference:

other: read across target

Reference

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study conducted on read-across material

Justification for type of information:

A RAAF report will shortly be provided.

Reason / purpose for cross-reference:

read-across source

Irritation parameter:

other: relative mean tissue viability

Run / experiment:

Mean

Value:

68.8

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

valid

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Qualifier:

no guideline followed

Principles of method if other than guideline:

Performed using the standard protocol with the SkinEthic Model (SkinEthic Reconstituted Human Corneal model (HCE, SkinEthic Laboratories, Nice, France))

GLP compliance:

yes (incl. QA statement)

Remarks:

Date of Signature: 15/10/2007; Date of Inspection: 21/08/2007

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of sodium hydrogen N-(1-oxooctadecyl)-L-glutamate and sodium hydrogen N-(1-oxohexadecyl)-L-glutamate

EC Number:

936-610-7

Molecular formula:

See remarks

IUPAC Name:

Reaction mass of sodium hydrogen N-(1-oxooctadecyl)-L-glutamate and sodium hydrogen N-(1-oxohexadecyl)-L-glutamate

Constituent 2

Reference substance name:

Reaction mass of sodium hydrogen N-(1-oxooctadecyl)-L-glutamate and sodium hydrogen N-(1-oxohexadecyl)-L-glutamate

IUPAC Name:

Reaction mass of sodium hydrogen N-(1-oxooctadecyl)-L-glutamate and sodium hydrogen N-(1-oxohexadecyl)-L-glutamate

Test animals / tissue source

Details on test animals or tissues and environmental conditions:

In-vitro study, so no test animals used.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: Triplicate tissues treated with 30 µl of Solution A served as the negative control and triplicate tissues treated with 30 µl of 1% w/v Sodium Dodecyl Sulphate served as the positive control.

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 30 mg
- Concentration (if solution): not applicable


VEHICLE
- Amount(s) applied (volume or weight with unit): not applicable
- Concentration (if solution): not applicable
- Lot/batch no. (if required): not applicable
- Purity: not applicable

Duration of treatment / exposure:

10 minutes

Observation period (in vivo):

Not applicable

Number of animals or in vitro replicates:

Triplicate SkinEthic tissues

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Rinsed with water.
- Time after start of exposure: 10 minutes


SCORING SYSTEM: relative mean viability


TOOL USED TO ASSESS SCORE: optical density was measured (quantitative measurement of tissue viability) at 540nm (OD540) using the Anthos 2001 microplate reader.

Results and discussion

In vitro

Results

Irritation parameter:

other: Relative mean tissue viability

Run / experiment:

Mean

Value:

ca. 68.8

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Any other information on results incl. tables

Assessment of Eye Irritation Potential – Viability of HCE Tissues

Material	Individual Tissue Viability	Mean OD540	% Viability
Negative ControlÅ	1.034	1.008	100*
	0.981
Positive ControlÅ	0.525	0.488	48.4
	0.451
Test Material	0.793	0.693	68.8
	0.593

* = The mean viability of the negative tissues is set at 100%

Å = Control group shared with another study

* = The mean viability of the negative tissues is set at 100%

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

According to the protocol followed the test material was considered to be a Non-Irritant (NI).

Executive summary:

Introduction. 

The purpose of this study was to determine the eye irritation potential of the test materials using the SkinEthic Reconstituted Human Corneal model (HCE, SkinEthic Laboratories) after a treatment period of 10 minutes. The test is based on the hypothesis that irritant chemicals are able to penetrate the corneal epithelial tissue and are sufficiently cytotoxic to cause cell death.

Methods. 

The experimental design of the study consists of a test for Direct Reduction of MTT by the test materials, followed by the main test.

For the main test, triplicate SkinEthic tissues were treated with 30 mg of the test material for 10 minutes. Triplicate tissues treated with 30 µl of Solution A served as the negative control and triplicate tissues treated with 30 µl of 1% w/v Sodium Dodecyl Sulphate served as the positive control.

At the end of the exposure period each SkinEthic tissue was rinsed. The rinsed tissues (two per group) were taken for MTT loading. The remaining tissues were retained for possible histopathology. Following MTT loading the reduced MTT was extracted from the tissues.

After extraction the absorbency of triplicate aliquots of the extracted MTT solution for each SkinEthic tissue was measured (OD₅₄₀). Data are presented in the form of % viability (MTT conversion relative to negative controls).

The test material was classified according to the following criteria:

i)                   If the % relative mean tissue viability was ≥ 60% the test material was considered to be Non-Irritant (NI).

ii)                  If the % relative mean tissue viability was < 60% the test material was considered to be Irritant (I).

Results.

The relative mean viability of the test material treated tissues after a 10 minute exposure was 68.8%.

It was considered unnecessary to proceed with tissue histopathology.

Quality criteria.

The quality criteria required for acceptance of results in the test were satisfied.

Conclusion.

According to the protocol followed the test material was considered to be a Non-Irritant (NI).