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-)

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:

2016

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)

Version / remarks:

adopted 28th July 2015

Deviations:

no

GLP compliance:

yes (incl. QA statement)

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

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

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

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.

Reference 2

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1991

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: meets generally accepted scientific standards and acceptable for assessment

Qualifier:

no guideline available

Principles of method if other than guideline:

The study report contains the limited information on the test conditions because the study has been performed in 1991. However available information is sufficient to conclude on the classification of the substance. Further the test was performed on the vertebrates and use of results from old experimental studies is one of the options to provide information requested by REACH. New experimental studies with vertebrates must only be conducted if there is no adequate existing information.

GLP compliance:

not specified

Species:

rabbit

Strain:

New Zealand White

Type of coverage:

not specified

Preparation of test site:

not specified

Vehicle:

water

Controls:

no

Amount / concentration applied:

TEST MATERIAL- Amount(s) applied (volume or weight with unit): 0.5 g per animal

Duration of treatment / exposure:

4 h

Observation period:

72 h

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

other: 24 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 48 h

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

other: 48 h

Score:

0

Max. score:

4

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

other: 72 h

Score:

0

Max. score:

4

Interpretation of results:

not classified

Conclusions:

Practically non-irritating to intact skin; very slightly irritating to scarified skin.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1991

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: meets generally accepted scientific standards and acceptable for assessment

Qualifier:

no guideline available

Principles of method if other than guideline:

The study report contains the limited information on the test conditions because the study has been performed in 1991. However available information is sufficient to conclude on the classification of the substance. Further the test was performed on the vertebrates and use of results from old experimental studies is one of the options to provide information requested by REACH. New experimental studies with vertebrates must only be conducted if there is no adequate existing information.

GLP compliance:

not specified

Species:

rabbit

Strain:

New Zealand White

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL- Amount(s) applied: 0.1 g

Duration of treatment / exposure:

no data

Observation period (in vivo):

72 h

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 24 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 h

Remarks on result:

other: 1/10 corneal surface opaque

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 72 h

Score:

0

Max. score:

4

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

other: 24 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 72 h

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

other: 72 h

Score:

0

Max. score:

3

Interpretation of results:

not classified

Conclusions:

Based on level of eye damage, the test substance, Reactive Violet 1, has not been classified.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Based on the skin and eye irritation/corrosion test results, the test substance is not irritating to eyes and skin.