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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.
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.
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