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Toxicological information

Skin irritation / corrosion

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Administrative data

Endpoint:
skin irritation / corrosion
Remarks:
other: in vitro (validated and accepted)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
16 January, 2012 - 20 January, 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study has been performed according to OECD and/or EC guidelines and according to GLP principles.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report date:
2012

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
other: EU Guideline B.40 BIS: "In Vitro Skin Corrosion: Human Skin Model Test".
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: OECD Guidelines for Testing of Chemicals, Guideline no. 431: In Vitro Skin Corrosion: Human Skin Model Test (adopted 13 April 2004).
Deviations:
no
GLP compliance:
yes

Test material

Constituent 1
Details on test material:
- Name of test material (as cited in study report): FAT 40854/A TE
- Substance type: Reddish-brown powder
- Physical state:solid
- Purity: 46.2% (4 main constituents)
- Lot/batch No.: TZ 5719 / BOP 02-11
- Expiration date of the lot/batch: 01 April 2016
- Storage condition of test material: At room temperature in the dark
- Hygroscopic: Yes, store in well-sealed container
- Volatile: No
- pH: 5.4 at concentration of >80g/L
- Stability in Water: Stability for at least 6 hours at room temperature is confirmed over the concentration range 20-200 mg/mL, NOTOX Project 497840
- Solubility in Water: More than 80 g/L

Test animals

Species:
human

Test system

Vehicle:
unchanged (no vehicle)
Amount / concentration applied:
TEST MATERIAL
- Amounts applied: 25 mg solid test substance, 25 μl Milli-Q water to moisten skin.

NEGATIVE CONTOL:
- Amount applied: 50 µl Milli-Q water

POSITIVE CONTROL
- Amount applied: 50 µl
- Concentration: 8N KOH
Duration of treatment / exposure:
3 minutes and 1 hour
Details on study design:
TEST SITE
- EpiDerm Skin Model (EPI-200, Lot no.:16222 kit W). The model consists of normal, human-derived epidermal keratinocytes which have been cultured to form a multilayered, 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 EpiDerm tissues (surface 0.6 cm²) were cultured on polycarbonate membranes of 10 mm cell culture inserts.

REMOVAL OF TEST SUBSTANCE
- Washing: phosphate buffered saline
- Time after start of exposure: 3 minutes and 1 hour

POST INCUBATION PERIOD
- Over night at room temperature

SCORING SYSTEM:
- Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) at the end of the treatment.
The amount of extracted formazan was determined spectrophotometrically at 540 nm in triplicate withthe TECAN Infinite® M200 Pro Plate Reader.
Cell viability was calculated for each tissue as percentage of the mean of the negative control tissues.
Skin corrosion potential of the test substance was classified according to remaining cell viability following exposure of the test substance with either of the two exposure times.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
other: tissue viability
Basis:
other: percentage of control
Time point:
other: 3 minutes
Score:
86
Remarks on result:
other: Negative control = 100%; Positive control = 17%
Irritation parameter:
other: tissue viability
Basis:
other: percentage of control
Time point:
other: 1 hour
Score:
95
Remarks on result:
other: Negative control = 100%; Positive control = 15%

Any other information on results incl. tables

FAT 40854/A TE was checked for possible direct MTT reduction by adding the test substance to MTT medium. Because no colour change was observed it was concluded that FAT 40854/A TE did not interact with MTT.

Applicant's summary and conclusion

Interpretation of results:
other: not corrosive
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
The in vitro skin irritation test was conducted according to OECD 431 guideline and GLP principles.
It is concluded that this test is valid and that the test substance is not corrosive in the in vitro skin corrosion test.
Executive summary:

In an in vitro skin corrosion test using a human skin model (EpiDerm (EPI-200)), the influence of the test substance on the viability of human skin was tested. The test substance was applied directly to 0.6 cm2 cultured skin (25 mg, in presence of 25 μl Milli-Q water). After 3 minutes and 1 hour, the substance was removed and cells were cultured over night. The viability of the cells was tested by reduction of MTT. Survival of unexposed skin was set at 100%, the positive control had a mean cell viability of 17% and 15% for the 3 -minute and 1 hour application respectively whereas the test substance showed cell viability of 86% and 95% for the 3 -minute and 1 hour application respectively.

Since the mean relative tissue viability after exposure to the test substance was not below 50% after 3 minutes treatment and not below 15% after 1 hour treatment, it can be concluded that the test substance is not corrosive in the in vitro corrosion test.