Spectral Unmixing-Based Reaction Monitoring of Transformations between Nucleosides and Nucleobases.
UV/Vis spectroscopy
nucleobases
nucleoside phosphorylase
nucleosides
spectral unmixing
Journal
Chembiochem : a European journal of chemical biology
ISSN: 1439-7633
Titre abrégé: Chembiochem
Pays: Germany
ID NLM: 100937360
Informations de publication
Date de publication:
14 09 2020
14 09 2020
Historique:
received:
01
04
2020
revised:
21
04
2020
pubmed:
24
4
2020
medline:
8
7
2021
entrez:
24
4
2020
Statut:
ppublish
Résumé
The increased interest in (enzymatic) transformations between nucleosides and nucleobases has demanded the development of efficient analytical tools. In this report, we present an update and extension of our recently described method for monitoring these reactions by spectral unmixing. The presented method uses differences in the UV absorption spectra of nucleosides and nucleobases after alkaline quenching to derive their ratio based on spectral shape by fitting normalized reference spectra. It is applicable to a broad compound spectrum comprising more than 35 examples, offers HPLC-like accuracy, ease of handling and significant reductions in both cost and data acquisition time compared to other methods. This contribution details the principle of monitoring reactions by spectral unmixing, gives recommendations regarding solutions to common problems and applications that necessitate special sample treatment. We provide software, workflows and reference spectra that facilitate the straightforward and versatile application of the method.
Identifiants
pubmed: 32324971
doi: 10.1002/cbic.202000204
pmc: PMC7540295
doi:
Substances chimiques
Nucleosides
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
2604-2610Informations de copyright
© 2020 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.
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