2,7-Naphthalenedisulfonic acid, 4-amino-6-[2-[4-[[[4-[2-(2,4-dihydroxyphenyl)diazenyl]phenyl]sulfonyl]amino]phenyl]diazenyl]-5-hydroxy-3-[2-(4-nitrophenyl)diazenyl]-, sodium salt (1:2)

Endpoint summary

Description of key information

not eye irritant for rabbits

not skin irritant for rabbits

Key value for chemical safety assessment

Skin irritation / corrosion

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Additional information

For skin irritation/corrosion:

Two tests are available on similar substance 1 and 2, both confirming that the substance is not skin irritant.

For eye irritation/corrosion:

Classification for corrosion on the similar EC: 284-407-0 was based on a study on the similar substance 1 presented in the framework of DSD notification of new substances, reporting eye damage not reversible within 28 days (Stahl); few information is available on the study and several attempts to recover the original reports have been made, without success, either by the owner of the notification or the responsible member state (France, presumably).

 

According to article 3.3.2.6 of CLP Regulation the proposed classification is justified in the following cases:

Case 1)

“These observations include animals with grade 4 cornea lesions and other severe reactions (e.g., destruction of cornea) observed at any time during the test, as well as persistent corneal opacity,discoloration of the cornea by a dye substance, adhesion, pannus, and interference with the function of the iris or other effects that impair sight. In this context, persistent lesions are considered those which are not fully reversible within an observation period of normally 21 days.”

Case 2)

Substances are also classified in Category 1 if, when applied to the eye of an animal, a substance produces:

–at least in one animal effects on thecornea, iris orconjunctiva that are not expected to reverse or have not fully reversed within an observation period of normally 21 days;

and/or

– at least in 2 of 3 tested animals, a positive response of:

– corneal opacity ≥ 3 and/or

– iritis > 1.5

calculated as the mean scores following grading at 24, 48 and 72 hours afterinstallation of the test material.

 

Case 1) doesn’t apply since the reports cite the following:

Coloration noire de la membrane nictitante réversible à J14 chez 2 animaux et non réversible à 28J chez 1 animal. Cette coloration a empeché l'appreciation de l'erytheme conjonctival jusque J14.

But discoloration is not referred to cornea.

Case 2) applies since the report indicates for conjunctiva score that for all animals the maximum value of redness has been observed at 28 days, while for all the other end points very slight effect has been reported.

Based on the available information the report is missing of many important details, like the identification of the substance and the related impurities and potential additives in case a commercial sample has been tested. No detail comments on controls and animal housing have been reported.

 

The result is in contradiction with the following complete studies performed on the similar substance 1 and on the other studies on similar substance 2

 

Study 1: Key study – reliability 2 – Read across study from similar substance 2

J. Zapatero (1997). PRIMARY EYE IRRITATION IN RABBITS TEST SUBSTANCE : CARBON DERMA AF 135. Test Substance: Carbon Derma AF 135. Testing laboratory: CENTRO DE INVESTIGATION Y DESAROLLO APLICADO S. A. L. Report no.: CD-96/5303T-1. Owner company: CLARIANT PRODUCTIONS S. A.

All the effects have been properly assessed and described in detail. No effect has been reported for any end point after 6 days in any animal

 

Study 2: Supporting study – reliability 2 – Read across study from similar substance 2

BASF AG (1984a). Report on the acute irritation to the eye of the white rabbit based on OECD 405. Testing laboratory: BASF Department of Toxicology. Report no.: 83/259. Owner company: BASF SE. Report date: 1984-03-01 (Original language: German).

 

The study has been rated Klimish 2 either because it has been performed on the similar substance 2 and it is not a GLP study even if the study is complete and performed according the OECD guideline 405 without deviations.

The substance is well identified and a certificate has been provided for exact composition.

All the effects have been properly assessed and described in detail. No effect has been reported for any end point after 72 hours in any animal.

Study 3: Supporting study – reliability 1 - Read across study from similar substance 1

L. Bisini (2014). ACID BLACK 210 Na, ASSESSMENT OF OCULAR CORROSION/SEVERE IRRITATION, BOVINE CORNEAL OPACITY/PERMEABILITY (BCOP) ASSAY. Testing laboratory: RTC. Report no.: 98470. Owner company: REACH & Colours Kft . (Original language: English).

In order to completely assess the end point the Lead Registrant REACH & Colours Kft performed an in vitro test according to OECD 437, on similar substance 1, which composition has been well defined.

 

The BCOP test method is recommended as an initial step within a tiered-testing strategy to identify chemicals inducing serious eye damage, i.e. chemicals to be classified as UN GHS Category 1, without further testing. A chemical that is not predicted as causing serious eye damage or as not classified for eye irritation/serious eye damage with the BCOP test method would require additional testing (in vitro and/or in vivo) to establish a definitive classification

According to the updated OECD 437 guideline of 17 September 2012, the IVIS cut-off values for identifying test chemicals as inducing serious eye damage (UN GHS Category 1) and test chemicals not requiring classification for eye irritation or serious eye damage (UN GHS No Category) are given hereafter:

  

IVIS	UN GHS
< 3	No Category
>3; < 55	No prediction can be made
> 55	Category 1

 

The test resulted in a medium IVIS of 25.3 and alterations of the mean cornea opacity were observed. However, since the test item was coloured, the mean opacity value is affected by the substance remaining on the corneal surfaces. No relevant increases in corneas permeability were recorded after treatment with the test item when compared to those of negative control.

 

Conclusions

Two Klimish 2 studies (Clariant 1997 and BASF 1984) have been performed on similar substance 2 showing no effect that can trigger to a classification. They are well performed, the substance is well identified and their results are consistent. An in vitro study, according to OECD 437 has been performed on the similar substance 1. The result according to the new update of OECD 437 of the 17 September 2012 cannot lead to a conclusion about the classification, but it can be considered as a weight of evidence, together with the two previous cited study.

Based on read across from similar 1 and 2, Acid Green 68 could be considered as a not-irritating substance.

Read across is discussed more in detail in the Read Across document in section 13.

Justification for classification or non-classification

No classification for skin irritation/corrosion is warranted under Regulation 1272/2008

No classification for eye irritation/corrosion is warranted under Regulation 1272/2008