Ashes (residues), vanadium-contg.

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

28.04.2010 - 05.05.2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GPL guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: OECD guideline 431

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Ashes (residues), vanadium-contg.

EC Number:

282-208-3

EC Name:

Ashes (residues), vanadium-contg.

Cas Number:

84144-87-6

Molecular formula:

Not available

Constituent 2

Reference substance name:

Vanadium concentrate

IUPAC Name:

Vanadium concentrate

Details on test material:

- Name of test material (as cited in study report): vanadium concentrate
- Preparation date: 07/2009
- Expiry date: 30. Sept. 2011
- Batch No.: 14-45810

Test animals

Species:

other: in vitro, human skin modell

Strain:

other: not applicable (in vitro membrane test)

Details on test animals or test system and environmental conditions:

Not applicable: in vitro membrane test with human skin model.

Cell line:
Reconstructed Human Epidermis (EPISKIN). Kit of 12 units, age day 17 (supplied by SkinEthics)

Preperation of skin model:
At arrival, human skin was inspected for color or agar medium: orange color was verified. The temperature indicator was also inspected to verify that kit has not been exposed to a temperature above 40 °C. After arrival, inserts of human skin model were transferred in 1 ml of Maintenance Medium and incubated for 3 h at 37 +- 1 °C at 5 % CO2.
All culture media and reagents are in balance with room temperature when in contact with human skin.

Test system

Type of coverage:

other: not applicable (in vitro membrane test)

Preparation of test site:

other: not applicable (in vitro membrane test)

Vehicle:

unchanged (no vehicle)

Controls:

yes

Amount / concentration applied:

Substance is used as it is.
For solid materials, sufficient test substance was applied evenly to cover the skin, and it was moistened with deionised or distilled water to ensure good contact with the skin.

Duration of treatment / exposure:

3 min and 1 h at 37+- 1 °C and 5 % CO2

Observation period:

not applicable

Number of animals:

not applicable (in vitro membrane test)

Details on study design:

Positive control: KOH 8 N (CAS: 1310-58-3); reference substance is used as it is.
Negative control: sterile distilled water

Treatment:
Three tissue replicates are used for each sample treatment, two for positive control and one for the negative one. For solid materials, sufficient test substance was applied evenly to cover the skin, and it was moistened with deionised or distilled water to ensure good contact with the skin. The plates were incubated for 3 min and 1 h at 37 +- 1 °C at 5% CO2

MTT cell viability evaluation:
At the end of the exposure period, the test material was washed with DPBS for twenty times from the skin surface. Every tissue replicates was treated in 0.3 ml of MTT solution 1 mg/ml. The plate was incubated for 3h at 37 +- 1 °C at 5 % CO2. At the end of incubation each tissue insert was treated with 2 ml of Isopropyl alcohol (dark, T room, overnight). Two replicates of extract solution (200 µl) were put into microplates's wells and the optical density (OD) of the extracted (solubilised) dye at 570 nm was evaluated.

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

other: other: MTT cell viability evaluation

Value:

98.54

Remarks on result:

other:

Remarks:

Basis: mean. Time point: 3 min treatment. Reversibility: other: not applicable. (migrated information)

Results 2

Irritation / corrosion parameter:

other: other: MTT cell viability evaluation

Value:

98.46

Remarks on result:

other:

Remarks:

Basis: mean. Time point: 1 h treatment. Reversibility: other: not applicable. (migrated information)

Any other information on results incl. tables

Table 1: Cell viability (%)

Contact time	3 min	1 h
KOH 8N	9.03	0.28
Sample	98.54	98.46

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

According to OECD 431 "In vitro Skin Corrosion Human Skin Model test" sample resulted not corrosive to the skin.

Executive summary:

Skin corrosive properties of Vanadium concentrate were assessed in an human skin model (EpiSKIN) according to OECD 431. Sufficient test substance was applied evenly to cover the skin, and it was moistened with distilled water to ensure good contact. The plates were incubated for 3 min and 1 h at 37 °C. Afterwards, a MTT cell viability test evaluation was conducted.

It was verified that the mean viability (respect negative control) of the two replicates of positive control was ≤ 20 % after 1h. The mean OD value of the two replicates of negative control was ≥ 0.8.

Vanadium concentrate resulted to be not corrosive to human skin.

This in vitro dermal study on human skin models is acceptable and satisfies the guideline requirement for a guideline study according to OECD 431 "In vitro Skin Corrosion Human Skin Model test".