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EC number: 911-351-2 | CAS number: -
Direct MTT Reduction:
The MTT solution containing the test item did not turn blue. This was taken to indicate the test item did not reduce MTT.
The relative mean tissue viability for the positive control treated tissues was 8.3% relative to the negative control treated tissues following the 240-minute exposure period. The positive control acceptance criterion was therefore satisfied. The mean OD540for the negative control treated tissues was 0.217. The negative control acceptance criterion was therefore satisfied.
Mean OD540of duplicate tissues
Relative mean viability (%)
Negative Control Item
Positive Control Item
*= The mean viability of the negative control tissues is set at 100% Diisobutyl hydrogen phosphate and Isobutyl dihydrogen phosphate (IBAP)
In an in vitro skin corrosion study performed according to the OECD guideline 431 and in compliance with the GLP, Reaction Mass of "Diisobutyl hydrogen phosphate and Isobutyl dihydrogen phosphate" (purity 96%) was applied undiluted to Reconstructed Human Epidermis (RHE) model (Episkin TM model kit 0.38cm²).
Duplicate tissues were treated with the test item for exposure periods of 3, 60 and 240 minutes. At the end of the exposure period each tissue was rinsed from the test item before each tissue was taken for MTT-loading. After MTT loading a total biopsy of each epidermis was made and placed into micro tubes containing acidified isopropanol for extraction of formazan crystals out of the MTT-loaded tissues. At the end of the formazan extraction period each tube was mixed thoroughly and duplicate 200 µl samples were transferred to the appropriate wells of a pre‑labelled 96‑well plate. The optical density (OD) was measured at 540 nm (OD540). 0.9 % (w/v) Sodium Chloride Solution was used as the negative control. Glacial Acetic Acid was used as the positive control.
Percentage of cell viability (MTT reduction in the test item treated tissues relative to negative control tissues) was determined for each exposure period. The relative mean viability of the test item treated tissues was 103.7; 11.1 and 4.1% for exposure period of 3, 60 and 240 min, respectively.
As viability of cells is higher than 35% after 3 minutes exposure but lower than 35% after 1 hour exposure the Reaction Mass of "Diisobutyl hydrogen phosphate and Isobutyl dihydrogen phosphate" is classified as corrosive to the skin, Skin Corr. 1B (H314) according to the Regulation (EC) 1272/2008 (CLP) and as C, R34 according to the Directive 67/548/EEC.
This study is considered as acceptable as it satisfies the main criteria of OECD guideline No. 431.
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