Separation of Nucleobases Using High-performance Liquid Chromatography Coupled with Voltammetric Scanning

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 0.05M borate buffer (pH9.0) as the mobile phase, at isocratic flow rate 1mLmin(-1), and at separation column temperature of 30 degrees C. The accurate timing of the cyclic voltammetry enabled us to quantify the concentrations of individual nucleobases by oxidation on glassy carbon electrode.