Butanedioic acid, sulfo-, C-C16-18-alkyl esters, disodium salt

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2008

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: INVITTOX (UK) protocol no. 98

Version / remarks:

1994

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Butanedioic acid, sulfo-, C-C16-18-alkyl esters, disodium salt

EC Number:

294-268-8

EC Name:

Butanedioic acid, sulfo-, C-C16-18-alkyl esters, disodium salt

Cas Number:

91697-07-3

Molecular formula:

C20H38O7S.2Na – C22H40O7S.2Na

IUPAC Name:

octasodium 4-(hexadecyloxy)-4-oxo-2-sulfonatobutanoate 4-(hexadecyloxy)-4-oxo-3-sulfonatobutanoate 4-(octadecyloxy)-4-oxo-2-sulfonatobutanoate 4-(octadecyloxy)-4-oxo-3-sulfonatobutanoate

Test material form:

solid

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: CS82130005
- Storage: Room temperature
- Expiry Date: 2009-07-30

Test animals / tissue source

Species:

cattle

Strain:

not specified

Details on test animals or tissues and environmental conditions:

- Test system: Freshly isolated bovine cornea
- Source: Odenwaldschlachthof Brensbach, D-64395, Brensbach

Test system

Vehicle:

Hank's balanced salt solution

Controls:

yes

Amount / concentration applied:

The test item was tested as a 5% suspension.

Duration of treatment / exposure:

10 minutes

Duration of post- treatment incubation (in vitro):

2 h

Number of animals or in vitro replicates:

3 corneas per group

Details on study design:

SELECTION AND PREPARATION OF CORNEAS
All eyes were carefully examined macroscopically for defects. Those presenting defects such as vascularization, pigmentation, opacity and scratches were discarded.
The cornea was carefully removed from the eye using scalpel and rounded scissors. A rim of about 2mm of tissue (sclera) was left for stability and handling of the isolated cornea. All corneas used in the experiment were collected in Hank's BSS supplemented with streptomycin / penicillin and checked finally with a view box for defects listed above.
Each cornea was mounted in a cornea holder with the endothelial side against the sealing ring (o-ring) of the posterior part of the holder. The cornea was gently flattened over the O-ring but stretching was avoided. After the anterior part of the holder was positioned on top of the cornea and fixed in place with srews, both compartments of the holder were filled with complete medium. The posterior compartment was filled first to return the cornea to its natural convex position. Care was taken to assure no air bubbles were present within the compartments.

NUMBER OF REPLICATES: 3

NEGATIVE CONTROL USED
0.9 % saline

POSITIVE CONTROL USED
2-Ethoxyethanol

APPLICATION DOSE AND EXPOSURE TIME
5% suspension, 10 min incubation

POST-INCUBATION PERIOD: yes
2h

REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period: minimum 3 times
- POST-EXPOSURE INCUBATION: 2h

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: yes
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of UV/VIS spectrophotometry

SCORING SYSTEM: In Vitro Irritancy Score (IVIS)

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

cornea opacity score

Run / experiment:

1st

Value:

-0.33

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

cornea opacity score

Run / experiment:

2nd

Value:

0.67

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

cornea opacity score

Run / experiment:

3rd

Value:

0.67

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Results 4

Irritation parameter:

other: In vitro Score

Run / experiment:

Mean

Value:

0.86

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Applicant's summary and conclusion

Interpretation of results:

study cannot be used for classification

Conclusions:

In conclusion, it can be stated that under the conditions of this study, the test item is not considered to be an eye irritant.

Executive summary:

The test substance (5% suspension) was tested in the in-vitro BCOP assay for its eye irritation properties. The test item was applied to corneas and incubated for 10 minutes at 32 °C. After incubation the corneas were washed and incubated for another 120 minutes. Opacity was measured before and after treatment and after postincubation time. After the opacity measurements permeability was determined with a fluorescein solution. With the negative control neither an increase of opacity nor permeability of the corneas could be observed. The positive control showed clear opacity and distinctive permeability of the corneas.

The test item did not cause any opacity or permeability of the corneas. In conclusion, it can be stated that under the conditions of this study, the test item is not considered to be an eye irritant.