Analytic separation techniques for carbohydrates - Chromatography of carbohydrates - High performance size exclusion chromatography (HPSEC)
5 important questions on Analytic separation techniques for carbohydrates - Chromatography of carbohydrates - High performance size exclusion chromatography (HPSEC)
What is HPSEC and how can it be used for carbohydrates?
It separates the carbohydrates based on hydrodynamic volume.
On what does the hydrodynamic volume depend?
- Number of sugar building blocks
- Type of linkages present
- Occurrence, type and length of side branches
- Occurrence and type of charge
What kind of information can you monitor with the PHSEC?
- processing
- enzymatic digestion
- fermentation
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How is the detection of carbohydrates done in HPSEC?
- refractive index (RI)
- Multi angles light scattering (MALLS)
- viscosity detectors
What is the conclusion about HPSEC and carbohydrates?
- Rapid technique for screening fate of polysaccharide upon processing, digestion, fermentation, etc
- Take care for aggregation/molecular interaction
Not all columns are suitable for all polymers (e.g. chitosan, xanthan, etc) - Hydrodynamic volume measured rather then Mw!
- Calibration preferably to be done with look-a-like molecules! Or using MALLS and RI detection! Viscosimetric detector aids on functional properties!
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