4-nitrotoluene-2-sulphonic acid

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:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)

Version / remarks:

2009

Principles of method if other than guideline:

Bovine Corneal Opacity and Permeability Test (BCOP-Test)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

4-nitrotoluene-2-sulphonic acid

EC Number:

204-445-3

EC Name:

4-nitrotoluene-2-sulphonic acid

Cas Number:

121-03-9

Molecular formula:

C7H7NO5S

IUPAC Name:

2-methyl-5-nitrobenzenesulfonic acid

Test system

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 750 µl of a 20% test-substance preparation

Details on study design:

Corneal opacity was measured quantitatively as the amount of light transmission through the cornea. Permeability was measured quantitatively as the amount of sodium fluorescein dye that passes across the full thickness of the cornea. Both measurements were used to calculate an In Vitro Irritancy Score (IVIS) of the test substance, which was used for prediction of serious eye damage.

Each treatment group (test substance, NC and PC) consisted of 3 corneas.

Before application the medium in the anterior chamber was removed using a syringe.

750 µL of the 20% (w/v) test-substance preparation (non-surfactant) was applied into the anterior chamber using a pipette.

Control tissues were concurrently applied into the anterior chamber with 750 µL of highly de-ionized water (negative control, NC) or with 750 µL of 20% (w/v) solution of Imidazole in highly de-ionized water (positive control, PC) using a pipette.

The corneas were incubated in a horizontal position at about 32 °C for approximately 4 hours (non-surfactant solids). The test substance, NC and PC were then removed from the anterior chamber using a syringe and the epithelium was washed at least 3 times with Eagle’s MEM (containing phenol red) and once with Eagle’s MEM (without phenol red). Both chambers were then refilled with fresh Eagle’s MEM (without phenol red).

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

in vitro irritation score

Value:

143.6

Vehicle controls validity:

not examined

Negative controls validity:

not specified

Positive controls validity:

not specified

Results 2

Irritation parameter:

cornea opacity score

Value:

143.4

Vehicle controls validity:

not examined

Negative controls validity:

not specified

Positive controls validity:

not specified

Any other information on results incl. tables

The BCOP test showed the following results: The mean opacity change of the test-substance treated corneas was 143.4. The mean OD490 of the test-substance treated corneas determined as permeability parameter was 0.013. The In Vitro Irritancy Score (IVIS) of the test-substance treated corneas was calculated to be 143.6.

Based on the observed results and applying the evaluation criteria it was concluded, that 4-Nitrotoluene-2-sulphonic acid causes serious eye damage in the Bovine Corneal Opacity and Permeability Test (BCOP Test) under the test conditions chosen.

Applicant's summary and conclusion

Interpretation of results:

other: classified in Category 1 according to the CLP Regulation EC No. 1272/2008

Conclusions:

The substance is an eye irritant.

Executive summary:

The corneal damage potential of the substance was assessed according to OECD guideline 437.
Corneal opacity was measured quantitatively as the amount of light transmission through the cornea. Permeability was measured quantitatively as the amount of sodium fluorescein dye that passes across the full thickness of the cornea. The mean Opacity change of the test-substance treated corneas was 143.4. The mean OD490 of the test-substance treated corneas determined as permeability parameter was 0.013. The In Vitro Irritancy Score (IVIS) of the test-substance treated corneas was calculated to be 143.6.