Tetrasodium [3-[(3-chloro-2-hydroxy-5-sulphophenyl)azo]-5-[[4-chloro-6-(3-sulphoanilino)-1,3,5-triazin-2-yl]amino]-4-hydroxynaphthalene-2,7-disulphonato(6-)]cuprate(4-)

Skin irritation / corrosion

Administrative data

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

adopted 28th July 2015

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Tetrasodium [3-[(3-chloro-2-hydroxy-5-sulphophenyl)azo]-5-[[4-chloro-6-(3-sulphoanilino)-1,3,5-triazin-2-yl]amino]-4-hydroxynaphthalene-2,7-disulphonato(6-)]cuprate(4-)

EC Number:

274-977-9

EC Name:

Tetrasodium [3-[(3-chloro-2-hydroxy-5-sulphophenyl)azo]-5-[[4-chloro-6-(3-sulphoanilino)-1,3,5-triazin-2-yl]amino]-4-hydroxynaphthalene-2,7-disulphonato(6-)]cuprate(4-)

Cas Number:

70880-03-4

Molecular formula:

C25H11Cl2CuN7O14S4.4Na

IUPAC Name:

tetrasodium [3-[(3-chloro-2-hydroxy-5-sulphophenyl)azo]-5-[[4-chloro-6-(3-sulphoanilino)-1,3,5-triazin-2-yl]amino]-4-hydroxynaphthalene-2,7-disulphonato(6-)]cuprate(4-)

Test material form:

solid

Details on test material:

Batch No.: 7004 Composition of test substance: - Reactive Violet 1 (CAS No. 70880-03-4) 85.0 % w/w - NaCl (CAS No. 7647-14-5) 15.0 % w/w

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Vehicle:

water

Details on test system:

The reconstructed human epidermal model EpiDerm (EPI-200, MatTek, Ashland, USA) consists of normal human epidermal keratinocytes which have been cultured to form a multilayered highly differentiated model of the human epidermis. The EpiDerm System is manufactured according to defend quality assurance procedures.The EpiDerm tissues (surface 0.63 cm2) are cultured on specially prepared cell culture inserts and shipped as kits containing 24 tissues on shipping agarose together with the necessary amount of culture media.

Control samples:

yes, concurrent vehicle

yes, concurrent positive control

Amount/concentration applied:

25 mg of the test substance was placed directly atop to the previously moistened tissue with 25 µL water. The material was spread on the tissue surface.

Duration of treatment / exposure:

3 min, 60 min

Duration of post-treatment incubation (if applicable):

3 hours

Number of replicates:

3

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

3 min

Value:

92.1

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Results 2

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

60 min

Value:

90.6

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Applicant's summary and conclusion

Interpretation of results:

other: not corrosive to skin

Conclusions:

Under experimental conditions, the test substance, Reactive Violet 1, is not corrosive in In vitro Skin Corrosion Test on EpiDerm tissues.

Executive summary:

The test substance, Reactive Violet 1,was assayed for the in vitro skin corrosion in human epidermal model EpiDerm. The test was performed according to OECD Test Guideline 431, In vitro Skin Corrosion: Human Skin Model Test, adopted in July 28^th, 2015.

Direct reduction test in test tubes was performed before MTT test. As the test substance is coloured violet, the test gave no information about direct MTT reduction. Potential direct reduction caused by rests of the test substance in tissues was excluded after the test in frozen tissues.

In MTT test, the test substance (25 mg) was placed directly atop to the previously moistened tissue. Length of exposition was 3 and 60 minutes. Nine tissues were used for the experiment in each time, three per test substance, three for positive control, three for negative control. After rinsing tissues were incubated with MTT for three hours and then extracted with isopropyl alcohol for two hours at room temperature with shaking. OD(570) of isopropyl alcohol extracts was measured on a spectrophotometer. Relative cell viability was calculated for each tissue as per cent of the mean viability of the negative control tissues.

Under experimental conditions, average reactivity of tissues treated by the test substance, Reactive Violet 1, was 92.1 % of the negative control after 3 minutes treatment and 89.6 % (after correction) after 60 minutes treatment. According the test conditions, the test substance is not corrosive in In vitro Skin Corrosion Test on EpiDerm tissues.