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Derivatize, Racemize, and Analyzean Easy and Simple Procedure for Chiral Amino Acid Standard Preparation for Enantioselective Metabolomics

Horak, Jeannie ; Lämmerhofer, Michael

Analytical chemistry (Washington), 2019-06, Vol.91 (12), p.7679-7689 [Periódico revisado por pares]

United States: American Chemical Society

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  • Título:
    Derivatize, Racemize, and Analyzean Easy and Simple Procedure for Chiral Amino Acid Standard Preparation for Enantioselective Metabolomics
  • Autor: Horak, Jeannie ; Lämmerhofer, Michael
  • Assuntos: Amino acids ; D-Amino acids ; Enantiomers ; High temperature ; High-performance liquid chromatography ; Isomers ; Liquid chromatography ; Metabolomics ; Mining ; N-Terminus ; Racemization ; Stereochemistry ; Theanine ; Urea ; Ureas
  • É parte de: Analytical chemistry (Washington), 2019-06, Vol.91 (12), p.7679-7689
  • Notas: ObjectType-Article-1
    SourceType-Scholarly Journals-1
    ObjectType-Feature-2
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  • Descrição: A simple, controllable, and reproducible stereoisomerization (racemization and epimerization) protocol for the preparation of scalemic α-amino acid mixtures from stereoisomerically pure standards was developed. Simply derivatize your amino acids with a racemization tag that incorporates a urea bond on the N-terminus of the target amino acid and incubate at elevated temperatures up to 95 °C for defined time periods until the targeted d-amino acid levels are obtained. The racemization tags investigated were 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC), aminophenyl-N-hydroxysuccinimidyl carbamate (AC), and 3-aminopyridyl-N-hydroxysuccinimidyl carbamate (APC). Employing this method, it was possible to create a ready-to-use, tailor-made chiral uniformly 13C and 15N labeled [U- 13C15N]-amino acid standard with the desired d-amino acid percentage within minutes or hours without sample cleanup. A racemization time of 30 min at 95 °C will lead to a d-amino acid level of 1–5%, while 6 h at 95 °C provides 15–30% d-amino acids. Racemization occurs due to imine formation at the chiral carbon atom bound to the urea-linking group without decomposition of labile amino acids such as Asn, Gln, Trp, Cit, and theanine. For amino acids possessing two chiral centers such as dl-Ile or dl-Thr, only the epimerization of isomers with different stereochemistry at the second chiral center will produce all four possible isobaric enantiomers. All measurements were performed on the zwitterionic Chiralpak ZWIX­(+) column using a dual hydro-organic flow gradient combined with HPLC-ESI-QTOF-MS analysis. This new racemization method solves the problem of (enantioselective) matrix effects and inaccurate results in LC-MS based enantioselective metabolomics and warrants full MS-compatibility.
  • Editor: United States: American Chemical Society
  • Idioma: Inglês
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  • Realização: ABCD-USP USP   Logos de Redes Sociais: Freepik

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