1-Naphthalenesulfonic acid, 3-diazo-3,4-dihydro-4-oxo-, ester with phenyl(2,3,4-trihydroxyphenyl)methanone 6-diazo-5,6-dihydro-5-oxo-1-naphthalenesulfonate

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

October 9 to 11, 2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2009

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

3‐benzoyl‐2,6‐bis({[3‐(‐lambda5‐diazynylidene)‐4‐oxo‐3,4‐dihydronaphthalen‐1‐yl]sulfonyl}oxy)phenyl 3‐(‐lambda5‐diazynylidene)‐4‐oxo‐3,4‐dihydronaphthalene‐1‐sulfonate

Cas Number:

11562-47-8

Molecular formula:

not applicable as UVCB

IUPAC Name:

3‐benzoyl‐2,6‐bis({[3‐(‐lambda5‐diazynylidene)‐4‐oxo‐3,4‐dihydronaphthalen‐1‐yl]sulfonyl}oxy)phenyl 3‐(‐lambda5‐diazynylidene)‐4‐oxo‐3,4‐dihydronaphthalene‐1‐sulfonate

Test material form:

solid

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: skin model obtained from Episkin/SkinEthic Laboratories, Lyon, France

Justification for test system used:

stansard model

Vehicle:

unchanged (no vehicle)

Details on test system:

CELL CULTURE
- Supplier: Episkin/SkinEthic Laboratories, Lyon, France
- Source: human keratinocytes cultured on a polycarbonate filter in conditions which permit their terminal differentiation
- Format: 24 well plate
- Batch: 19-RHE-146


TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment: room temperature
- Temperature of post-treatment incubation: 37°C

REMOVAL OF THE TEST MATERIAL AND CONTROL
After the end of the treatment interval, the residual test item was removed immediately by gently rinsing with a minimum volume of 25 mL DPBS using a pipette. Excess DPBS was removed by gently shaking the inserts and blotting the bottom with blotting paper.

DYE BINDING METHOD
- Dye used in the dye-binding assay: MTT
- Spectrophotometer:ELx800, BioTek Instruments GmbH, Bad Friedrichshall, Germany at 570 nm

Control samples:

yes, concurrent negative control

Amount/concentration applied:

TEST MATERIAL: 16 mg of solid test material
NEGATIVE CONTROL: 16 µL (Phosphate-Buffered Saline)
POSITIVE CONTROL: 16 µL (5% aqueous solution of sodium dodecyl sulfate in deionised water)

Duration of treatment / exposure:

42 min (± 1 minute)

Duration of post-treatment incubation (if applicable):

42 hours (± 1 hour)

Number of replicates:

3

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

run 1, mean of three tissues

Value:

95.3

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

OTHER EFFECTS:
- Visible damage on test system: none
- Direct-MTT reduction: none
- Colour interference with MTT: none

- Acceptability of the Quality Control Data of the Skin Model with Reference to Historical Batch Data:
The OD values for the negative control shall be in the range of ≥ 0.8 and ≤ 3.0 as given in OECD Guideline 439. The values obtained were: 1.714 to 1.779

- Acceptability of the Positive and Negative Control stated by Episkin/SkinEthic Laboratories:
The negative control data meet the acceptance criteria if the mean OD value of the 3 tissues is ≥ 1.2 at 570 nm. The standard deviation value is considered valid if ≤ 18% of group mean-value.
The results obtained were:
Mean OD = 1.757, SD =2.1%

The positive control data meet the acceptance criteria if the mean viability value, expressed as % of the negative control, is < 40%. The standard deviation value is considered valid if ≤ 18% of group mean-value.
Mean OD = 0.026, SD =6.7%, mean viability: 1.5 %

Any other information on results incl. tables

 

Group	Time / [min]	Mean OD	Mean Relative viability / [%]
Negative Control	42	1.757	100
Positive Control	42	0.026	1.5
Test Material	42	1.675	95.3

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of the present study, the test item is not considered to possess an irritant potential to skin (UN GHS: No Category).