Effect of Acid on Separation of Xanthines

Primesep B separates xanthines such as theobromine, dimethylxanthine, theophylline by a combination of reversed-phase and ion-exchange mechanisms. The anion-exchange properties of the column allow retention of hydrophilic xanthines, and the reversed-phase properties retain hydrophobic compounds. The HPLC separation uses a mobile phase of water, acetonitrile (MeCN, ACN) and either trifluoroacetic acid (TFA), perchloric acid (HClO4), or sulfuric acid (H2SO4) and UV detection.


Column Primesep B
Mobile Phase MeCN/H2O
Buffer TFA , HClO4, H2SO4
Flow Rate 1.0 ml/min


Class of Compounds Drug, Acid, Hydrophilic, Ionizable, Hormone
Analyzing Compounds Theobromine, 1,7 - Dimethylxanthine, Theophyline
Application Analytes:
Theophylline monohydrate