Disodium; 4-Amino-3-{4-[4-(2,4-diamino-phenylazo)-benzenesulfonylamino]-phenylazo}-5-hydroxy-6-phenylazo-naphthalene-2,7-disulfonate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2020

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2020

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

disodium 4-amino-3-[2-(4-{4-[2-(2,4-diaminophenyl)diazen-1-yl]benzenesulfonamido}phenyl)diazen-1-yl]-5-hydroxy-6-(2-phenyldiazen-1-yl)naphthalene-2,7-disulfonate

EC Number:

605-104-5

Cas Number:

157577-99-6

Molecular formula:

C34H26N10Na2O9S3

IUPAC Name:

disodium 4-amino-3-[2-(4-{4-[2-(2,4-diaminophenyl)diazen-1-yl]benzenesulfonamido}phenyl)diazen-1-yl]-5-hydroxy-6-(2-phenyldiazen-1-yl)naphthalene-2,7-disulfonate

Test system

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

The test item was tested as suspension prepared from test item at 20% concentration in a 0.9% sodium chloride solution.
1g of the test item was suspended in 5 mL of 0.9% sodium chloride solution

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Remarks on result:

not determinable

Other effects / acceptance of results:

The value of IVIS for the positive control (20% imidazole in 0.9% NaCl) obtained during the study was 73.28. This value is within the acceptance limit (one standard deviation of the current historical mean), so the study is considered acceptable.
The value of opacity for negative control (0.9% NaCl) obtained during the study was 1.89
and value of permeability was 0.050.
The values obtained during this study not exceeded upper limits, so the study is considered acceptable.
The basic values for the IVIS calculation (opacity values) could not be measured because of colouring of corneas which were applied by the test item.
Therefore the In Vitro Irritancy Score (IVIS) for test item, Acid Black 234, could not be calculated.
For this reason the classification was not performed

Any other information on results incl. tables

Group	Cornea No.	Appearance after exposure
Negative control (0.9% NaCl)	1	Without macroscopic damage
	2	Without macroscopic damage
	4	Without macroscopic damage
Positive control (20% Imidazole in 0.9% NaCl)	4	Corneal opacity
	5	Corneal opacity
	6	Corneal opacity
Test item (Acid Black 234)	10	colouring by the test item – marked  (black colour)
	11	colouring by the test item – marked  (black colour)
	12	colouring by the test item – marked  (black colour)

Applicant's summary and conclusion

Interpretation of results:

study cannot be used for classification

Conclusions:

The basic values for the IVIS calculation (opacity values) could not be measured because of colouring of corneas which were applied by the test item.
Therefore the In Vitro Irritancy Score (IVIS) for test item, Acid Black 234, could not be calculated.
For this reason the classification was not performed

Executive summary:

The test item, Acid Black 234, was tested for the potential to cause ocular corrosivity or severe irritancy, as measured by its ability to induce opacity and increased permeability in an isolated bovine cornea. The test was performed according to OECD Test Guideline No. 437 (2017) and Method B.47 Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage, Commission Regulation (EU) 2017/735, Adopted 14th February 2017.

The testing was performed on three groups of corneas: test item treatment group (20% in NaCl), positive control group (20% w/v imidazole in 0.9% NaCl solution) and negative control group (0.9% NaCl solution). Three corneas per group were used.

The closed-chamber method was used, because the test item solution was applicable by micropipette. The opacity and permeability of each cornea were measured. The In Vitro Irritancy Score (IVIS) was calculated from the values of opacity and permeability.

The opacity and permeability of each cornea were measured. The In Vitro Irritancy Score (IVIS) was calculated from the values of opacity and permeability.

The basic values for the IVIS calculation (opacity values) could not be measured because of colouring of corneas which were applied by the test item.
Therefore the In Vitro Irritancy Score (IVIS) for test item, Acid Black 234, could not be calculated.
For this reason the classification was not performed.