Registration Dossier

Administrative data

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
30 August 2010 - 4 October 2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted under GLP conditions and OECD Guideline 431

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: OECD Guideline 431: In Vitro Skin Corrosion: Human Skin Model Test (adopted April 13, 2004)
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): MTDID 21721, TFEE-5
- Substance type: clear colorless liquid
- Physical state: liquid
- Lot/batch no.: TFEE5-276/12/09-1
- Expiration date of the lot/batch: 12 Dec 2012
- Stability under test conditions: stable
- Storage condition of test material: room temperature in the dark under nitrogen

Test animals

Species:
human
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: EpiDerm Skin Model, MatTek Corporation, Ashland, MA, USA
- Age at study initiation:
- Weight at study initiation:
- Fasting period before study:
- Housing:- Diet (e.g. ad libitum):
- Water (e.g. ad libitum):
- Acclimation period:
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 37.0 +/- 1.0
- Humidity (%): 73 - 86
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light):
IN-LIFE DATES: From: 30 August 2010 To: 3 September 2010

Test system

Type of coverage:
other: undiluted test substance was added to the cell culture plates on top of the skin tissues
Preparation of test site:
other: Skin samples were transferred to plates containing DMEM medium and incubated for 80 min at 37C in 5% CO2, Medium was replaced prior to test article application.
Vehicle:
unchanged (no vehicle)
Controls:
other: One tissue sample was treated with 50 uL of Milli-Q water (negative control) and one was treated with 50 uL of 8N KOH (positive control).
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 50 uL/well
- Concentration (if solution): undiluted
VEHICLE
- Amount(s) applied (volume or weight with unit):
- Concentration (if solution):
- Lot/batch no. (if required):
- Purity:
Duration of treatment / exposure:
3 min (two tissues) and 1 h (two tissues)
Number of animals:
4 tissue samples were treated with the test material
Details on study design:
TEST SITE
- Area of exposure:
- % coverage:
- Type of wrap if used:
REMOVAL OF TEST SUBSTANCE
- Washing (if done):
- Time after start of exposure:
SCORING SYSTEM: Cell viability was calculated as the percentage of the mean optical density at 540 for the negative control tissue. A test substance is considered corrosive if cell viability compared to the negative control is below 50% after 3 min exposure or below 15% after 1 h exposure.

Results and discussion

In vitro

Resultsopen allclose all
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
3 minute exposure
Value:
ca. 100
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other:
Remarks:
Basis: other: The relative mean tissue viability after 3-min exposure to TFEE-5 compared to the negative control tissue was 100%.
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
1 hour exposure
Value:
ca. 97
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other:
Remarks:
Basis: other: The relative mean tissue viability after 1 hour exposure to TFEE-5 compared to the negative control tissue was 97%.

Any other information on results incl. tables

TFEE-5 was checked for possible direct MTT reduction and none was observed. The cell viability (optical density at 540 nm) for the negative control was within the laboratory historical control range. The mean viability for the positive control following 3 min of exposure was 9%. The maximum inter-tissue variability in viability for tissues treated identically was less than 11% and the maximum difference between the mean viability of two tissues and one of the two tissues was less than 6%. It is therefore concluded that the test system functioned properly.

Applicant's summary and conclusion

Interpretation of results:
other: not corrosive
Remarks:
Criteria used for interpretation of results: other: OECD Test Guideline 431 criteria
Conclusions:
TFEE-5 was not corrosive to the skin in this test system
Executive summary:

OBJECTIVE: The skin corrosion potential of TFEE-5 (MTDID 21721, batch TFEE5-276/12/09-1) was evaluated in an in vitro three dimensional human epidermal skin model (EpiDerm EPI-200). METHODS: This study was conducted in accordance with OECD GLP (1997). The study design was based on OECD 431 (2004) and EC 440/2008, Part B.40 (2008). The test material was tested as received. TFEE-5 was tested for possible direct MTT reduction prior to the study and was shown to not interact with MTT. The in vitro skin was placed in 6-well plates with DMEM media underneath and incubated at 37 C in 5% CO2. TFEE-5 (50 uL) was added to two tissues for a 3 minute exposure and two tissues for a 1 hour exposure. Negative control (Milli-Q water) and positive control (8 N potassium hydroxide) were tested concurrently in a similar manner. After the exposure period the tissues were washed with phosphate buffered saline (PBS) to remove residues. The tissues were then incubated in the presence of MTT for 3 hours at 37 C in a 5% CO2 atmosphere. The tissues were washed with PBS and formazan was extracted with isopropanol overnight at room temperature. The amount of extracted formazan was determined spectrophotometrically at 540 nm. Cell viability was calculated as percentage of the mean of the negative control tissues. A test material is considered corrosive if the relative mean tissue viability is <50% following a 3 minute exposure or <15% following a 1 hour exposure. RESULTS: The negative and positive controls performed as expected which indicated the test was valid. Relative mean tissue viability was 100% and 97% following the 3 minute and 1 hour exposures to TFEE-5, respectively. CONCLUSION: Based on the results of this study, TFEE-5 was not corrosive in the in vitro skin corrosion test.