1H-Benz[de]isoquinoline-1,3(2H)-dione, 6-amino-, N,N'-bis[mixed 2-ethylhexyl and 3-[(2-ethylhexyl)oxy]propyl] derivs

Endpoint summary

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

16/05/2017-18/06/2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)

Qualifier:

according to guideline

Guideline:

other: Council regulation 440/2008, Method B.40 BIS:"In vitro skin Corrision:Human skin model Test", May 30, 2008

Qualifier:

according to guideline

Guideline:

other: MatTek Corporation protocol for: In Vitro EpiDerm TM Skin Corrosion Test (EPI-200-SCT) For Use with MatTek Corporation's reconstructed Human Epidermal Model EpiDerm; Version 07/11/2014.

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- batch No.of test material: 78-242-15
- Expiration date of the batch: 01/02/2018

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature, protected from light
- Stability under test conditions: not applicable

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
The test item was applied undiluted. 50 µL of the test item was dispensed directly atop the EpiDerm TM tissue. The test item was spread to match size of the tissue. A nylon mesh was used as spreading aid.

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

foreskin from multiple donors

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

after 3 minutes

Value:

>= 50

Vehicle controls validity:

not specified

Negative controls validity:

valid

Positive controls validity:

not specified

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

after 60 minutes

Value:

>= 15

Vehicle controls validity:

not specified

Negative controls validity:

valid

Positive controls validity:

not specified

The test item showed no non-specific MTT-reducing potential but it showed colouring after mixture with isopropanol. Therefore, NSC_livingwas determined. No correction of results was necessary because NSC_livingwas below 5% (3 min treatment: 0.3%; 60 min treatment: 0.6%).

The test item showed no corrosive effects. The mean relative tissue viability (% negative control) was³ 50% (104.7%) after 3 min treatment and³ 15% (93.8%) after 60 min treatment.

Interpretation of results:

GHS criteria not met

Conclusions:

In this study under the given conditions the test item showed no corrosive effects. the test item is classified as "non-corrosive".

Executive summary:

In the present study the skin corrosivity potential of the test item was analysed. Since corrosive chemicals are cytotoxic after a short time exposure to the stratum corneum of the epidermis the cytotoxic effects of the test item on EpiDermÔ, a reconstituted three-dimensional human epidermis model, were determined. Hereby, the test item was applied topically. Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT after a 3 min and 60 min exposure period and compared to those of the concurrent negative controls.

The test item showed no non-specific MTT-reducing potential but it showed colouring after mixture with isopropanol. Therefore, NSC_livingwas determined. No correction of results was necessary because NSC_livingwas below 5% (3 min treatment: 0.3%; 60 min treatment: 0.6%).

The test item showed no corrosive effects. The mean relative tissue viability (% negative control) was³ 50% (104.7%) after 3 min treatment and³ 15% (93.8%) after 60 min treatment.

Reference 2

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

22/11/2017-15/12/2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Qualifier:

according to guideline

Guideline:

EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)

Qualifier:

according to guideline

Guideline:

other: MatTek Corporation Protocol for: In Vitro EpiDermTM Skin Irritation Test (EPI-200-SIT) For use with MatTek Corporation’s Reconstructed Human Epidermal Model EpiDerm (EPI-200-SIT); Version 07-Nov-2014

GLP compliance:

yes (incl. QA statement)

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

foreskin from multiple donors

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

after 60 minutes treatment and 42 h post-incubation

Value:

> 50

Vehicle controls validity:

not specified

Negative controls validity:

valid

Positive controls validity:

not specified

The test item showed no irritant effects. The mean relative tissue viability (% negative control) was > 50% (104.2%) after 60 min treatment and 42 h post-incubation.

Interpretation of results:

GHS criteria not met

Conclusions:

In this study under the given conditions the test item showed no irritant effects. The relative mean tissue viability after 60 min of exposure and 42 h post-incubation was > 50%. The test item is therefore classified as “non-irritant” in accordance with UN GHS “No Category”.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

21/06/2017-21/06/2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Species:

other: Bovine

Strain:

not specified

Vehicle:

physiological saline

Amount / concentration applied:

TEST MATERIAL
It was not possible to get a 20% concentration of solution/suspension with the test item in physiological saline 0.9% NaCl. Therefore the test item was administered directly and moistened with physiological saline 0.9% NaCl.

VEHICLE
Physiological saline 0.9% NaCl (AlleMan Pharma, lot no. 609709, expiry date: 21/06/2017)

Number of animals or in vitro replicates:

3

Irritation parameter:

cornea opacity score

Value:

13.09

Vehicle controls validity:

not specified

Negative controls validity:

valid

Positive controls validity:

valid

Conclusions:

No prediction can be made regarding the classification of the test substance YELLOW 2747 according to the evaluation criteria. Further testing in another suitable method is required.

Reference 2

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

11/12/2017-13/12/2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 492 (Reconstructed Human Cornea-like Epithelium (RhCE) Test Method for Identifying Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage)

Qualifier:

according to guideline

Guideline:

other: EURL ECVAM DB-ALM Method Summary No. 164: EpiOcular™ Eye Irritation Test - Summary, 22 July 2015

Qualifier:

according to guideline

Guideline:

other: EpiOcular™ Eye Irritation Test (OCL-200-EIT) For the prediction of acute ocular irritation of chemicals For use with MatTek Corporation’s Reconstructed Human EpiOcular™ Model, 29 June 2015

GLP compliance:

yes (incl. QA statement)

Irritation parameter:

in vitro irritation score

Run / experiment:

The mean relative tissue viability

Value:

<= 60

Vehicle controls validity:

not specified

Negative controls validity:

valid

Positive controls validity:

valid

The controls confirmed the validity of the study. The mean absolute OD₅₇₀of the two negative control tissues was > 0.8 and < 2.5 (1.661). The mean relative tissue viability (% negative control) of the positive control was < 50% (18.6%). The inter tissue difference of replicate tissues of all dose groups was < 20% (18.6%).

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

In this study under the given conditions the test item showed irritant effects.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Justification for classification or non-classification

Skin irritation

The In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) test and the In Vitro Skin Irritation: Reconstructed Human Epidermis Test has been performed.

The test item showed no corrosive or irritant effects.

Eye irritation

The eye irritancy potential of YELLOW 2747 was investigated in the bovine corneal opacity and permeability assay.

The result ( Mean in vitro irritation score) has been 13.09 .

The IVIS cut-off values for identifying test substances as inducing serious eye damage (UN GHS Category 1) and test substances not requiring classification for eye irritation or serious eye damage (UN GHS No Category) are:

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

The substance does not require classification for serious eye damage (UN GHS No Category), but no prediction can be made and further testing is required.

The In vitro Eye Irritation: Ocular Irritation Assay using the EpiOcular^TMHuman Tissue Model has been performed.

In this study under the given conditions the test item showed irritant effects. The test item is classified as “irritant“ in accordance with UN GHS “Category 2”.