HPLC Separation of Adenosine and Adenine Using the Hydrogen Bonding Method

    Application Notes: Nucleosides are glycosylamines consisting of a nucleobase linked to a ribose or a deoxyribose sugar. Nucleoside are building blocks for DNA and RNA. These compounds are very polar in nature and contain groups available for hydrogen bonding interactions. Method for separation of adenine and adenosine were developed using a hydrogen-bonding method. There is a strong correlation between retention time for adenine/adenosine and the mobile phase composition, which consists of acetonitrile and methanol. Order of elution for compounds depends on the amount of acetonitrile and methanol.  Furthermore, ellution of adenine and adenosine can be reversed based on the composition of the mobile phase. Our method is compatible with LC/MS and preparative chromatography.


Column Sharc 1, 3.2x100 mm, 5 µm, 100A
Mobile Phase MeCN/MeOH
Buffer AmFm, Formic acid
Flow Rate 1.0 ml/min
Detection UV, 270 nm


Class of Compounds Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements
Analyzing Compounds Adenosine, Adenine
Application Analytes: