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Toxicological information

Skin irritation / corrosion

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Administrative data

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2009
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study performed under GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2009
Report date:
2009

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
other: OECD 431 in vitro Skin Corrosion: Human Skin Model Test
Deviations:
no
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Reference substance name:
Slags, steelmaking, elec. furnace (carbon steel production - EAF C)
IUPAC Name:
Slags, steelmaking, elec. furnace (carbon steel production - EAF C)
Constituent 2
Reference substance name:
Slags, steelmaking, elec. furnace
EC Number:
294-410-9
EC Name:
Slags, steelmaking, elec. furnace
Cas Number:
91722-10-0
Molecular formula:
not applicable
IUPAC Name:
294-410-9
Details on test material:
Name Slags, steelmaking, elec. furnace (carbon steel production - EAF C)
Appearance solid
Molecular formula not applicable (UVCB)
Molecular weight not applicable (UVCB)
Purity 100 w/w % slag
Homogeneity homogenous
Vapour pressure extremely low (melting point >300°C)
Stability solid slag is stable at room temperature
Solubility slightly soluble in water
Production date not stated (2009)
Expiry date 12/2024
Storage Room Temperature: (20 ± 5°C)

Test animals

Species:
other: Homo sapiens (skin)
Strain:
other: not applicable
Details on test animals or test system and environmental conditions:
not applicable since Human Skin Model Test was performed. This test uses commercially available Epi-200-Kit.
The EpiDermTM tissue consists of normal, human-derived epidermal keratinocytes which have been cultured to form a multi-layered, highly differentiated model of the human epidermis. It consists of organized basal, spinous, and granular layers, and a multi-layered stratum corneum containing intercellular lamellar lipid layers arranged in patterns analogous to those found in vivo. The EpiDermTM tissues are cultured on specially prepared cell cultures inserts.
Epi-200-Kit and MTT-100 assays diluent were obtained from MatTek Corporation in Ashland, USA.

Test system

Type of coverage:
other: in vitro
Preparation of test site:
other: not applicable
Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
25 mg of the test item/well
Duration of treatment / exposure:
1 h
Observation period:
3 min or 1 h
Number of animals:
not applicable
Details on study design:
The following media were obtained from MatTek Corporation (description from LAUS GmbH)
MTT Medium
Contains 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide, which can be re-duced to a blue formazan
PBS-Buffer
Solution for the rinsing of the tissues
Assay Medium
Serum-free DMEM medium
Test was performed in several well-plates to account for positive controls (KOH) and negative controls (deionised water) (The exact conduct of the test is described in the original study report)
For each experiment (3 min, 1 h), one 24-well-plate is prepared as holding plate. 12 wells of each plate are filled with 300 µL assay medium, the other 12 with 300 µL MTT medium. One additional plate is left empty. The plates are stored in the incubator.
After pre-incubation, the assay medium is replaced by fresh assay medium and the test is started, using two wells as negative control with 50 µL H2O demineralised, two wells as positive controls with 50 µL potassium hydroxide solution and two other wells for testing the test item.
After the respective incubation time, the inserts are removed from the plates and are thoroughly rinsed with PBS, blotted and set into the respective holding plate, using the wells containing assay medium. After transfer of all inserts, they are immediately moved to the wells containing MTT solution, blotting the bottom again before setting the insert into the MTT well. The tissues are incubated with MTT medium for three hours and the MTT medium is replaced by PBS buffer. This is then replaced several times. At last, each insert is thoroughly dried and set into the empty, pre-warmed 24-well-plate. Into each well, 2 mL isopropanol are pipetted. The plate is then left to stand over night at room temperature. The inserts in which formazan has been produced over night are extracted.
From each well, three replicates with 200 µL solution (each) are pipetted into a 96-well-plate which is read in a plate spectral photometer at 570 nm.

Results and discussion

In vitro

Resultsopen allclose all
Irritation / corrosion parameter:
other: other: % formazan formation
Value:
92.9
Remarks on result:
other:
Remarks:
Basis: other: Human Skin Model. Time point: 3 min. Max. score: 100.0. Reversibility: other: not applicable. (migrated information)
Irritation / corrosion parameter:
other: other: % formazan formation
Value:
97.5
Remarks on result:
other:
Remarks:
Basis: other: Human Skin Model. Time point: 1 h. Max. score: 100.0. Reversibility: other: not applicable. (migrated information)

In vivo

Irritant / corrosive response data:
no corrosive response
Formazan Production by skin in slag and controls
Slag Positive Control Incubation period
92.9% 26.9% 3 min
97.5% 14.0% 1 hour
Other effects:
no effects caused by slag

Any other information on results incl. tables

Absorption Values

Negative Control

Test Item

Positive Control

Incubation

Tissue 1

Tissue 2

Tissue 1

Tissue 2

Tissue 1

Tissue 2

 

1.850

1.855

1.769

1.712

0.508

0.493

3 min

1.838

1.865

1.753

1.701

0.498

0.494

1.894

1.875

1.758

1.687

0.528

0.490

1.748

1.819

1.703

1.782

0.233

0.252

1 hour

1.751

1.803

1.700

1.781

0.253

0.260

1.757

1.812

1.681

1.779

0.251

0.252

Mean

Mean

Mean

 

1.863

1.730

0.502

3 min

1.782

1.738

0.250

1 hour

  Calculated formazan production (% of controls)

Test Item

Positive Control

Incubation

92.9%

26.9%

3 min

97.5%

14.0%

1 hour

Validity criteria were fulfilled:

The negative control (deionised water) was within the normal range (compared with the historical data of the laboratory).

The positive control (8 mol/L KOH) showed corrosive effects on the test system.

Historical controls

Parameter

Optical Density Negative Control

Optical Density Negative Control

Formazan production
Positive Control

Formazan production
Positive Control

Incubation Time

3 minutes

1 hour

3 minutes

1 hour

Mean

1.823

1.761

25.5%

13.8%

Standard
Deviation

0.261

0.300

4.3%

1.6%

Range

1.512 – 2.134

1.377 – 2.133

20.5 – 34.4%

11.3 – 15.8%

Applicant's summary and conclusion

Interpretation of results:
other: not corrosive
Remarks:
Criteria used for interpretation of results: OECD GHS
Conclusions:
EAF C: Human Skin Model Test (in vitro): not corrosive
Executive summary:

To determine the skin corrosion potential of slags, steelmaking, elec. furnace (carbon steel production - EAF C), fine ground EAF C was tested in the Human Skin Model Test following OECD 431. One valid experiment was performed.

Two tissues of the human skin model EpiDermTM were treated with fine-ground EAF S for 3 min and 1 h, respectively. 25 mg of the solid test item were applied to each tissue, wetted with H2O and spread to match the tissue size. Deionised water was used as negative control. 8 mol/L KOH was used as positive control.

After treatment with the negative control, the absorbance values were well above the required acceptability criterion of mean OD > 0.8 for both treatment intervals thus demonstrating the metabolic activity of the tissues. The positive control showed clear corrosive effects for both treatment intervals.

After three minutes treatment with the slag, the relative absorbance values were reduced to 92.9 %. This value is well above the threshold for corrosion potential (50%). After one hour treatment, relative absorbance values were decreased to 97.5 %. This value, too, is well above the threshold for corrosion potential (15%).

Therefore, fine-ground slag, steelmaking, elec. furnace (carbon steel production - EAF C), is not corrosive in the Human Skin Model Test.