HPLC Separation of Sugar Phosphates

Sugar phosphates occur in biological systems. Sugar phosphates are parts of two metabolic pathways: glycolisis and pentose phosphate pathways. One of the purposes of sugar phosphates is to store and transfer energy in cells. Because of the presence of sugar fragment and phosphate moiety these molecules are very polar in nature. Mixed-mode chromatography was applied for separation of glucose and fructose phosphates and diphosphates. Compounds are separated based on anion-exchange properties with diphosphates retaining longer than mono phosphates. Monitoring is done by ELSD.    


Column Primesep SB, 4.6x150 mm, 5 µm, 100A
Mobile Phase MeCN/H2O - 20/80%
Buffer AmFm - pH 3.0
Flow Rate 1.0 ml/min
Detection ELSD


Class of Compounds Sugars
Analyzing Compounds Sodium, Glucose 6- phosphate, Fructose 6 –phosphate, Fructose 1.6 biphosphate, Ribulose 1.5 biphosphate
Application Analytes:
Fructose 1,6 Bisphosphate
Fructose 6-Phosphate
Glucose 6-Phosphate
Ribulose 1,5 Bisphosphate