HPLC Separation of Glucose 6- Phosphate and Glucose 1- Phosphate on Newcrom B Column

Separation type: Liquid Chromatography Mixed-mode

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High Performance Liquid Chromatography (HPLC) Method for Analysis of Glucose 6- Phosphate and Glucose 1- Phosphate

Structural isomer separation poses a challenging task when using reverse-phase chromatography. However both glucose variants, which only differ to which carbon the phosphate group is attached to, can be separated in HPLC isocratically on a reverse-phase mixed-mode anion-exchange Newcrom B column. What differentiates, the Newcrom B phase from other silica-based Analysis of glucose phosphate is important because glucose-6-phosphate is the product of the first step in glycolysis metabolic pathway and glucose-1-phosphate is the first step in glycogenolysis. Glucose-6-phosphate is converted to glucose-1-phosphate and vice versa by the phosphoglucomutase enzyme in biological systems.


Column Newcrom B, 4.6x150 mm, 5 µm, 100A
Mobile Phase MeCN/H2O - 50/50%
Buffer Formic Acid -  0.5%
Flow Rate 1.0 ml/min
Detection ELSD, 40 °C  MS- compatible mobile phase


Class of Compounds Hydrophilic,  Sugar
Analyzing Compounds Glucose 6- Phosphate,  Glucose 1- Phosphate
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Application Analytes:
Glucose 1- Phosphate
Glucose 6-Phosphate