LCMS Method for Analysis of Glucosamine-1-Phosphate and Glucosamine-6-Phosphate on Primesep S  Column

HPLC Method for Analysis of Glucosamine 6-phosphate, Glucosamine-1-Phosphate on Primesep S by SIELC Technologies

HPLC Method for Analysis of Glucosamine 6-phosphate, Glucosamine-1-Phosphate on Primesep S Column by SIELC Technologies


 High Performance Liquid Chromatography (HPLC) Method for Analysis of Glucosamine 6-phosphate, Glucosamine-1-Phosphate

Glucosamine-1-phosphate and Glucosamine-6-phosphate are two different phosphorylation forms of glucosamine, a naturally occurring amino sugar that is part of the structure of certain glycosaminoglycans like chondroitin sulfate and hyaluronic acid.

Glucosamine-1-Phosphate:

It is the phosphorylated form of glucosamine at the first carbon atom.
It is often involved in metabolic pathways that lead to the synthesis of UDP-glucose, which is a precursor in the synthesis of glycogen and glycosaminoglycans.
It can also be an intermediate in the synthesis of amino sugars.
Glucosamine-6-Phosphate:

This form is phosphorylated at the sixth carbon atom.
It is an important intermediate in the hexosamine biosynthetic pathway, which is involved in the formation of UDP-N-acetylglucosamine (UDP-GlcNAc), a key substrate for the biosynthesis of glycosaminoglycans and glycoproteins.
It plays a role in cellular signaling and regulation, especially in the context of insulin resistance and other metabolic processes.
Both forms are critical in various biochemical pathways, particularly in the synthesis of complex carbohydrates and in maintaining cellular functions, including joint and connective tissue health.

Glucosamine-1-phosphate and Glucosamine-6-phosphate can be retained, separated and analyzed using an Primesep S mixed-mode stationary phase column. The analysis employs a gradient method with a simple mobile phase consisting of water, acetonitrile (MeCN), and formic acid as a buffer. Detection is achieved using LC MS.

ColumnPrimesep S, 3.2 x 100 mm, 5 µm, 100 A
Mobile PhaseGradient MeCN/H2O – 80/20 – 50/50 %, 10 min
BufferFormic Acid – 1.0%
Flow Rate0.5 ml/min
DetectionSIM 258 -, SIM 260 +
Sample0.5 mg/ml
Injection volume1 µl
LOD*150 ppb
* LOD was determined for this combination of instrument, method, and analyte, and it can vary from one laboratory to another even when the same general type of analysis is being performed.

Class of Compounds
Amino sugars
Analyzing CompoundsGlucosamine 6-phosphate, Glucosamine-1-Phosphate

Application Column

Primesep S

Column Diameter: 3.2 mm
Column Length: 100 mm
Particle Size: 5 µm
Pore Size: 100 A

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Application Analytes:
Glucosamine 6-phosphate
Glucosamine-1-Phosphate

Application Detection:
LC MS Detection
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.