A Protocol for GC-MS Profiling of Chiral Secondary Amino Acids

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F60077344%3A_____%2F25%3A00604733" target="_blank" >RIV/60077344:_____/25:00604733 - isvavai.cz</a>

Výsledek na webu

<a href="https://link.springer.com/protocol/10.1007/978-1-0716-4334-1_11" target="_blank" >https://link.springer.com/protocol/10.1007/978-1-0716-4334-1_11</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1007/978-1-0716-4334-1_11" target="_blank" >10.1007/978-1-0716-4334-1_11</a>

Jazyk výsledku

angličtina

Název v původním jazyce

A Protocol for GC-MS Profiling of Chiral Secondary Amino Acids

Popis výsledku v původním jazyce

A simple analytical workflow is described for gas chromatographic-mass spectrometric (GC-MS)-based chiral profiling of secondary amino acids (AAs) in biological matrices. The sample preparation is carried out directly in aqueous biological sample extracts and involves in situ heptafluorobutyl chloroformate (HFBCF) derivatization-liquid-liquid microextraction of nonpolar products into hexane phase followed by subsequent formation of the corresponding methylamides from the HFB esters by direct treatment with methylamine reagent solution. The (O, N) HFB-butoxycarbonyl-methylamide AA products (HFBOC-MA) are separated on a Chirasil-L-Val capillary column and quantitatively measured by GC-MS operated in selected ion monitoring (SIM) mode. The protocol includes 12 simple pipetting steps and covers the quantitative analysis of 8L, D pairs of secondary amino acids, including proline, isomeric 3-, 4-hydroxyprolines, pipecolic acid, nipecotic acid, azetidine-2-carboxylic acid, and cis- and trans-5-hydroxy-L-pipecolic acid using 13C5L-proline as an internal standard. The individual analytical steps are commented on and explained, with emphasis on the chiral GC-MS analysis of secondary amino acids in human urine, serum, and peptide hydrolysate samples.

Název v anglickém jazyce

A Protocol for GC-MS Profiling of Chiral Secondary Amino Acids

Popis výsledku anglicky

A simple analytical workflow is described for gas chromatographic-mass spectrometric (GC-MS)-based chiral profiling of secondary amino acids (AAs) in biological matrices. The sample preparation is carried out directly in aqueous biological sample extracts and involves in situ heptafluorobutyl chloroformate (HFBCF) derivatization-liquid-liquid microextraction of nonpolar products into hexane phase followed by subsequent formation of the corresponding methylamides from the HFB esters by direct treatment with methylamine reagent solution. The (O, N) HFB-butoxycarbonyl-methylamide AA products (HFBOC-MA) are separated on a Chirasil-L-Val capillary column and quantitatively measured by GC-MS operated in selected ion monitoring (SIM) mode. The protocol includes 12 simple pipetting steps and covers the quantitative analysis of 8L, D pairs of secondary amino acids, including proline, isomeric 3-, 4-hydroxyprolines, pipecolic acid, nipecotic acid, azetidine-2-carboxylic acid, and cis- and trans-5-hydroxy-L-pipecolic acid using 13C5L-proline as an internal standard. The individual analytical steps are commented on and explained, with emphasis on the chiral GC-MS analysis of secondary amino acids in human urine, serum, and peptide hydrolysate samples.

Druh

C - Kapitola v odborné knize

CEP obor

—

OECD FORD obor

10406 - Analytical chemistry

Projekt

<a href="/cs/project/GA23-06600S" target="_blank" >GA23-06600S: Nové přístupy v cílené metabolomické analýze nestabilních metabolitů s keto skupinou: značení kvarterní amoniovou skupinou a stabilními izotopy</a><br>

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2025

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název knihy nebo sborníku

Metabolomic Profiling: Methods and Protocols, Methods in Molecular Biology

ISBN

978-1-0716-4333-4

Počet stran výsledku

15

Strana od-do

205-219

Počet stran knihy

286

Název nakladatele

Humana Press

Místo vydání

New York, NY

Kód UT WoS kapitoly

001555691800012