Formaldehyde, reaction products with m-phenylenediamine, sodium sulfide (Na2S) and sulfur

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

05-Jun-2012 to 11 June 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Formaldehyde, reaction products with m-phenylenediamine, sodium sulfide (Na2S) and sulfur

EC Number:

274-986-8

EC Name:

Formaldehyde, reaction products with m-phenylenediamine, sodium sulfide (Na2S) and sulfur

Cas Number:

70892-34-1

Molecular formula:

not applicable

IUPAC Name:

Formaldehyde, reaction products with m-phenylenediamine, sodium sulfide (Na2S) and sulfur

Test material form:

semi-solid (amorphous): gel

Remarks:

migrated information: paste

Details on test material:

- Name of test material (as cited in study report): C.I. Leuco Sulphur Brown 37
- Substance type: Dark brown paste
- Physical state: Solid
- Storage condition of test material: At room temperature in the dark

Test system

Amount / concentration applied:

TEST MATERIAL
- Amount applied: C.I. Leuco Sulphur Brown 37 was applied and incubated with a cotton swab directly on top of the skin tissue

NEGATIVE CONTOL:
- Amount applied: 25 µL Phosphate buffered saline

POSITIVE CONTROL
- Amount applied: 25 µL
- Concentration: 5% (aq) Sodium dodecyl sulphate

Duration of treatment / exposure:

Exposure: 15 minutes
Post incubation period: 42 hours

Details on study design:

TEST SITE
- Area of exposure: human epidermis model
- % coverage: 0.38 cm2

REMOVAL OF TEST SUBSTANCE
- Washing: phosphate buffered saline
- Time after start of exposure: 15 minutes

POST INCUBATION PERIOD
- 42 hours

SCORING SYSTEM:
- After a 42 hour incubation period, determination of the cytotoxic (irritancy) effect was performed. Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT at the end of the treatment. Cell viability was calculated for each tissue as a percentage of the mean of the negative control tissues.

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Remarks:

percentage of control / time point: 15 minutes

Run / experiment:

1

Value:

102

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

C.I. Leuco Sulphur Brown 37 is non-irritant in the in vitro skin irritation test

Executive summary:

The study was performed to investigate the skin irritation properties of the test substance according to OECD 439. The in vitro tissue model EPISKIN-SM was used as the test system. 5% (aq) Sodium dodecyl sulphate served as positive control and phosphate buffered saline as the negative control. The mean tissue viability was determined at 15 min after application of test item or the controls.

The positive control had a mean cell viability of 4% after 15 minutes exposure. The absolute mean OD570 (optical density at 570 nm) of the negative control tissues was within the laboratory historical control data range. The standard deviation value of the percentage viability of three tissues treated identically was less than 5%, indicating that the test system functioned properly.

Skin irritation is expressed as the remaining cell viability after exposure to the test substance. The relative mean tissue viability obtained after 15 minutes treatment with C.I. Leuco Sulphur Brown 37 compared to the negative control tissues was 102%. Since the mean relative tissue viability for C.I. Leuco Sulphur Brown 37 was above 50% after 15 minutes treatment C.I. Leuco Sulphur Brown 37 is considered to be non-irritant.