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Separation of Nucleobases Using High‐performance Liquid Chromatography Coupled with Voltammetric Scanning
Author(s) -
Zitka Ondrej,
Kudr Jiri,
Labuda Jan,
Xhaxhiu Kledi,
Adam Vojtech
Publication year - 2018
Publication title -
electroanalysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.574
H-Index - 128
eISSN - 1521-4109
pISSN - 1040-0397
DOI - 10.1002/elan.201800175
Subject(s) - nucleobase , guanine , chromatography , cyclic voltammetry , thymine , high performance liquid chromatography , analyte , chemistry , cytosine , voltammetry , analytical chemistry (journal) , electrode , dna , electrochemistry , nucleotide , biochemistry , gene
Abstract In this paper, a method based on chromatographic separation of analytes and their quantification using on‐line cyclic voltammetry is reported. The method based on high performance liquid chromatography on reverse phase column was optimized using free nucleobases‐guanine, adenine, thymine, and cytosine. The optimal separation of nucleobases was detected when using 0.05 M borate buffer (pH 9.0) as the mobile phase, at isocratic flow rate 1 mL min −1 , and at separation column temperature of 30 °C. The accurate timing of the cyclic voltammetry enabled us to quantify the concentrations of individual nucleobases by oxidation on glassy carbon electrode.