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

Separation type: Liquid Chromatography Mixed-mode


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.

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

Condition
Column Newcrom B, 4.6×150 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

Description
Class of Compounds Hydrophilic,  Sugar
Analyzing Compounds Glucose 6- Phosphate,  Glucose 1- Phosphate

Application Column

Newcrom B

The Newcrom columns are a family of reverse-phase-based columns. Newcrom A, AH, B, and BH are all mixed-mode columns with either positive or negative ion-pairing groups attached to either short (25 Å) or long (100 Å) ligand chains. Newcrom R1 is a special reverse-phase column with low silanol activity.

Select options
Application Analytes:
Glucose 1- Phosphate
Glucose 6-Phosphate
SIELC Technologies usually develops more than one method for each compound. Therefore, this particular method may not be the best available method from our portfolio for your specific application. Before you decide to implement this method in your research, please send us an email to research@sielc.com so we can ensure you get optimal results for your compound/s of interest.