Carbohydrates / Polysaccharides - Structure elucidation - Sugar composition analysis
8 important questions on Carbohydrates / Polysaccharides - Structure elucidation - Sugar composition analysis
How can you hydrolyse a neutral and water soluble polysacharde?
TFA is a acid that is volitile so it easy to use. If you wand to get rit of it you just let it stand and it will be gone in no time.
How does sugar composition analysis go?
- Converting polysaccharides into a mixture of monosaccharides by acid hydrolysis
- Analyze monosaccharides by:
- Gas chromatography
- HPLC
- Uronic acids analyzed by color assay or by HPAEC after methanolysis
Is a alfa or beta linkage storonger and
is a pyranose or furanose stornger and
is a soluble or insoluble polysacharide?
pyranose is stronger
insoluble is stronger (harder to hydroyse)
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Will a galactose rhamnose or a rhamnose galactose be more strong?
How can you hydrolyse uronic acid cotntaining materials?
2M HCL in mehtanol for 16 hours at 80 degrees
How does do a high performance anion-exchange chromatography (HPAEC) for a sugar?
How can you hydrolyse a fibrous material?
folowed by 1 MH2SO4 for 3 hours at 100 degrees.
Lot of salt is added. This is not always plesen for analysis.
After complete hydrolysis, the obtained monosaccharides can be analyzed by GC or HPLC, what are the limitations and advantages?
- Components must be volatile -> monosaccharides should be made volatile. (mono-sugars are reduced to their corresponding alditols, + acetylation step)
- Preparation is fast compared to HPLC
HPAEC:
- neutral sugars are negatively charged under alkaline condition
- Hydrolysis can be checked in 1 run
- Sample preparation is easy
- Analysis time is fast
- Pulsed amperometric detector (PAD) enables separation and quantification of monosaccharides with low detection limits
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