Study of disulfide reduction and alkyl chloroformate derivatization of plasma sulfur amino acids using gas chromatography–mass spectrometry

0376118 - BC 2013 RIV DE eng J - Journal Article
Švagera, Z. - Hanzlíková, D. - Šimek, Petr - Hušek, Petr
Study of disulfide reduction and alkyl chloroformate derivatization of plasma sulfur amino acids using gas chromatography–mass spectrometry.
Analytical and Bioanalytical Chemistry. Roč. 402, č. 9 (2012), s. 2953-2963. ISSN 1618-2642. E-ISSN 1618-2650
R&D Projects: GA MZd NS9755; GA ČR GAP206/10/2401; GA ČR GA203/09/2014
Institutional research plan: CEZ:AV0Z50070508
Keywords : plasma amino acids * disulfide-reducing agents * trichloroacetic acid deproteinization
Subject RIV: CB - Analytical Chemistry, Separation
Impact factor: 3.659, year: 2012
http://www.springerlink.com/content/781wln3085q3v21r/

Four disulfide-reducing agents, dithiothreitol (DTT), 2,3-dimercaptopropanesulfonate (DMPS), and the newly tested 2-mercaptoethanesulfonate (MESNA) and tris(hydroxypropyl)phosphine (THP), were investigated in detail for release of sulfur amino acids in human plasma. After protein precipitation with trichloroacetic acid (TCA), the plasma supernatant was treated with methyl, ethyl, or propyl chloroformate and the products were subjected to GC–MS analysis. All the tested agents proved to be rapid and effective reducing agents for the assay of plasma thiols. When compared with DTT, the novel reducing agents DMPS, MESNA, and THP provided much cleaner extracts and improved analytical performance. Validation performed with the THP agent and propyl chloroformate derivatization demonstrated the robustness and reliability of this simple sample-preparation methodology.
Permanent Link: http://hdl.handle.net/11104/0208610