Sodium 4-vinylbenzenesulphonate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

22 June 1992

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1992

Report date:

1992

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 438 (Isolated Chicken Eye Test Method for Identifying Ocular Corrosives and Severe Irritants)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Sodium 4-vinylbenzenesulphonate

EC Number:

220-266-3

EC Name:

Sodium 4-vinylbenzenesulphonate

Cas Number:

2695-37-6

Molecular formula:

C8H8O3S.Na

IUPAC Name:

.

Test animals / tissue source

Species:

chicken

Strain:

other: ROSS, spring chickens

Details on test animals or tissues and environmental conditions:

Approximately 7 week old, male or female chickens (ROSS, spring chickens), bodyweight range approximately 2.5 -3.0 kg, were used as eye-donors. Twelve heads of these animals were obtained from poultry slaughter house v.d.Bor, Amersfoortseweg 118, Nijkerkerveen, the Netherlands. Heads of the animals were severed immediately after sedation of the animals by electric shock and incision of the neck for bleeding, and before they reached the next station or the process line.
The heads were placed in small plastic boxes (3 heads per box) on a bedding of paper tissues moistened with isotonic saline.
Next, they were transported to the testing facility. During transportation, the heads were kept at ambient temperature.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

The test substance was applied in amounts of 0.03g, by powdering the entire surface of the cornea.

Duration of treatment / exposure:

total exposure period of 10 seconds

Duration of post- treatment incubation (in vitro):

The control eye and test eyes were examined at 30, 75, 120, 180 and 240 minutes after treatment

Number of animals or in vitro replicates:

The test substance was tested on five out of the six eyes; the sixth eye was treated in a similar way with isotonic saline only and served as a control.

Details on study design:

Within 2 hours after kill, eyes were carefully dissected and placed in a superfusion apparatus using the following procedure:
First the eye-lids were carefully removed without damaging the cornea and a small drop of Fluorescein sodium BP 2%w/v was applied to the corneal surface for a few seconds and subsequently rinsed off with isotonic saline of ambient temperature.
Next, the head with the fluorescein-treated cornea was examined with a slit-lamp microscope (Slit-lamp 900BM), to ensure that the cornea was not damaged. If undamaged, the eye was further dissected from the head without damaging the eye or cornea. Care was taken to remove the eye-ball from the orbit without cutting off the optic nerve too short.

The enucleated eye was placed in a stainless steel clamp with the cornea positioned vertically and transferred to a chamber of the superfusion apparatus. The clamp holding the eye was positioned in such a way that the entire cornea was supplied with isotonic saline from a bent, stainless steel tube, at a rate of ca 0.10 -0.15ml/min. The six chambers of the superfusion apparatus as well as the saline were temperature controlled at 32± 1.5°C.

Six eyes were selected and after placing in the superfusion apparatus, the eyes were examined again with the slit-lamp microscope to ensure that they were not damaged. Corneal thickness was measured using the Depth Measuring Attachment no.II for the Haag-Streit slit-lamp microscope. Thickness of the cornea was expressed in instrument units. An accurate measurement was taken at the corneal apex of each eye. Eyes with a corneal thickness deviating more than 10% of the average corneal thickness of the six eyes, or eyes that were unacceptably stained with fluorescein (score higher than 0.5), indicating the cornea to be permeable, or eyes that showed any other signs of damage, were rejected as test eyes and were replaced.

After an equilibration period of 45 -60 minutes, the corneal thickness of the six eyes was measured again to determine the zero reference value for corneal swelling calculations. At time t=0, i.e.immediately after the zero reference measurement, the test substance was applied to the eye. For this purpose, the clamp holding the eye was placed on a paper tissue outside the chamber with the cornea facing upwards. The test substance was applied in amounts of 0.03g, by powdering the entire surface of the cornea. After a total exposure period of 10 seconds, the corneal surface was rinsed thoroughly with 20ml of isotonic saline of ambient temperature. Next, the eye in the holder was returned to its chamber. This procedure was repeated for each test eye. The test substance was tested on five out of the six eyes; the sixth eye was treated in a similar way with isotonic saline only and served as a control. The control eye and test eyes were examined at 30, 75, 120, 180 and 240 minutes after treatment. All examinations were carried out with the slit-lamp microscope.

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

cornea opacity score

Run / experiment:

Timepoint: 30 min

Value:

1

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

not applicable

Results 2

Irritation parameter:

cornea opacity score

Run / experiment:

Timepoint: 75 min

Value:

> 1.3 - < 1.5

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

not applicable

Results 3

Irritation parameter:

cornea opacity score

Run / experiment:

Timepoint: 120 min

Value:

> 1.3 - < 1.5

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

not applicable

Results 4

Irritation parameter:

cornea opacity score

Run / experiment:

Timepoint: 180 min

Value:

> 1.3 - < 1.5

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

not applicable

Results 5

Irritation parameter:

cornea opacity score

Run / experiment:

Timepoint: 240 min

Value:

> 19.4 - < 24.1

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

not applicable

Results 6

Irritation parameter:

percent corneal swelling

Run / experiment:

Timepoint: 30 min

Value:

> 12.1 - < 15.5

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

not applicable

Results 7

Irritation parameter:

percent corneal swelling

Run / experiment:

Timepoint: 75 min

Value:

> 14.5 - < 19

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

not applicable

Results 8

Irritation parameter:

percent corneal swelling

Run / experiment:

Timepoint: 120 min

Value:

> 18 - < 24.1

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

not applicable

Results 9

Irritation parameter:

percent corneal swelling

Run / experiment:

Timepoint: 180 min

Value:

> 18.7 - < 22.4

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

not applicable

Results 10

Irritation parameter:

percent corneal swelling

Run / experiment:

Timepoint: 240 min

Value:

> 19.4 - < 24.1

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

not applicable

Applicant's summary and conclusion

Interpretation of results:

irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

On the basis of the results obtained with this ex vivo bioassay and according to the scheme for EC classification applied it can be concluded that Spinomar NaSS is irritating, but not corrosive to eyes.

Executive summary:

The eye irritation was assessed in vitro (ex vivo) by means of the Chicken Enucleated Eye Test according to a method equivalent to the OECD Guideline 438 Isolated Chicken Eye Test Method) in compliance with GLP. Five enucleated eyes were treated with the undiluted substance and one control eye was treated in the same manner except without the application of test material.

Application of the substance caused moderate coreneal swelling, slight to moderate corneal opacity and moderate fluorescein retention in the test eyes.

On the basis of the results obtained with this ex vivo bioassay and according to the scheme for EC classification applied it can be concluded that Spinomar NaSS is irritating, but not corrosive to eyes.