Simultaneous Determination of Purine and Pyrimidine Bases Using High-Performance Liquid Chromatography with Electrochemical Detection: Application to DNA Assay

Abstract
A sensitive method for simultaneous determination of purine and pyrimidine bases was developed using high performance liquid chromatography with electrochemical detection (ECD). Pyrimidines as well as purines were detectable by ECD under alkaline conditions, and the order of susceptibility to electrochemical oxidation was guanine > adenine > thymine > cytosine. These bases were separated on a vinyl alcohol copolymer gel column and determined by ECD. The detection limit for each base was picomole level. This method was applied to the quantitation of bases in the acid hydrolysates of DNA and oligodeoxy-nucleotides.