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Glycan
Glycans - Analysis of Oligo- and Polysaccharides, Glycoproteins and other related Substances
Glycans are polysaccharides, i.e. large organic molecules that consist of many individual glycosidically linked monosaccharide building blocks. In the literature, the term "glycan" also refers to certain carbohydrate parts of glycoconjugates, e.g. glycoproteins or glycolipids. Well-known substances that are glycans include cellulose, chitin and heparin. Carbohydrates and their conjugates are of enormous physiological importance in plants, animals and humans.
Glycans are often analysed using HPLC-(MS), whereby different separation techniques can be used depending on the analyte.
Applications
Separation of procainamide-labelled dextran standards on AMT HALO Glycan
The HALO® GLYCAN core-shell columns from AMT are HPLC columns specially developed for analysing glycans. They are ideal for analysing oligosaccharides and other carbohydrate residues derived from glycoproteins and proteoglycans, for example. The functional group is a novel ligand containing 5 hydroxyl groups, which are bound to the fused-core silica particles via a special linker. Ideally, these columns are used in HILIC separation mode.
Test conditions
- Column: 2.1 x 150 mm, HALO 2.7 μm glycan (92922-705)
- Mobile phase A: 50 mM ammonium formate, pH 4.45
- Mobile phase B: acetonitrile
- Gradient: 80-55% B in 25 min
- Flow rate: 0.6 mL/min.
- Pressure: 190 bar
- Temperature: 60°C
- Detection: UV 300 nm
- Injection volume: 3 μL sample
- Sample solvent: 70/30 ACN/water
- Response time: 0.5 sec.
- Data rate: 3.3 Hz
- Flow cell: 2.5 μL semi-micro
- LC system: Shimadzu Nexera
Glycan separation with Sepax Proteomix SAX-NP3
Sepax offers the Proteomix SAX HPLC columns for the separation of various glycans, glycoproteins and other substances using ion exchange chromatography.
Test conditions
Column: Proteomix SAX-NP3 50x4.6mm (403NP3-4605)
Mobile phase A: 80/20 water/acetonitrile
Mobile phase B: 80/20 250 mM ammonium formate, pH 4.5/acetonitrile
Gradient:
Time / min | %B |
1 | 0 |
5.55 | 65 |
5.65 | 100 |
7.65 | 100 |
7.75 | 0 |
14 | 0 |
Flow rate: 1 mL/min
Detector: Fluorescence, excitation 260 nm, emission 430 nm
Injection volume: 3 µL
Sample: N-Linked Glycans Bovine Fetuin (6.7 pmol/µL)
Downloads
Manufacturer overview
AMT
HPLC - HILIC:
- HALO Glycan
Sepax
HPLC - IEX:
- Proteomix SAX-NP10
- Proteomix SAX-NP3
- Proteomix SAX-NP5
HPLC - SEC:
- Zenix SEC-150, 3µm
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