N,O-Bis(trimethylsilyl)trifluoroacetamide is used as a derivatization reagent for gas chromatography and mass-spectrometry analysis of phenolic acids in fruits. It is also used as a reactant in the preparation of pyrimidinone ribosides and analogs which shows anti-tumor properties. It plays as an important role as a reagent for the regioselective desulfation of polysaccharide sulfates. Furthermore, it is used to prepare trimethylsilyl ethers of carbohydrates and alcohols.

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