Profiling of urinary amino-carboxylic metabolites by in-situ heptafluorobutyl chloroformate mediated sample preparation and gas chromatography-mass spectrometry

0458358 - BC 2017 RIV NL eng J - Journal Article
Hušek, Petr - Švagera, Z. - Hanzlíková, D. - Řimnáčová, Lucie - Zahradníčková, Helena - Opekarová, Iva - Šimek, Petr
Profiling of urinary amino-carboxylic metabolites by in-situ heptafluorobutyl chloroformate mediated sample preparation and gas chromatography-mass spectrometry.
Journal of Chromatography A. Roč. 1443, April 22 (2016), s. 211-232. ISSN 0021-9673. E-ISSN 1873-3778
R&D Projects: GA ČR GA13-18509S
Institutional support: RVO:60077344
Keywords : metabolite profiling * metabolomics * urine
Subject RIV: CE - Biochemistry
Impact factor: 3.981, year: 2016
http://www.sciencedirect.com/science/article/pii/S0021967316302692

A novel heptafluorobutyl chloroformate reagent (HFBCF) was examined for in-situ derivatization of amino-carboxylic metabolites in human urine. The sample preparation protocol provides cleaner extracts than other urinary GC–MS based methods. Reaction products of 153 urinary metabolites with HFBCF and two internal standards were investigated in detail by GC–MS and LC–MS. The new method was validated for 132 metabolites in artificial urine with special attention to potential disease biomarker candidates. The sample preparation protocol was finally evaluated by means of a certified organic acid standard mixture in urine and by GC–MS analysis of 100 morning urines obtained from healthy patients, where 112 physiological metabolites were quantified in a 25 µL sample aliquot. Most metabolites were found within the range reported in the reference human metabolome (HMDB) database and literature. The described method has been a promising tool for targeted GC–MS based metabolomic analysis in urine.

Permanent Link: http://hdl.handle.net/11104/0258630