http://www.mdpi.com/1420-3049/15/8/5162/pdf

I think this ll be helpful little bit

This PDF has clear explanation of dextrins

Dextrins are produced by the incomplete hydrolysis of starch or glycogen. There are many of combinations of D-Glc linked by α-(1→4) or α-(1→6) glycosidic bonds, with two, thre, four... units of Glc. So, I don't have an idea of how to measure the concentration of dextrins when that is a mixture, maybe if you have an standard concentration of various of dextrins. You can measure a concentration of glucose after you completely hydrolyse starch.

Can use the library or need to get the standard for comparison

To know accurate concentration of dextrin, we normally hydrolyze dextrin to glucose by several enzymes and estimate by glucose kit. It is quite same after HPLC separation, normally they are separated by the molecular weight (degree of polymerization DP), so you just divide the glucose concentration by DP, it is becoming to the actual concentration of dextrins.

Dear Kiyohiko Igarashi,

i) Can you explain a little bit on why by hydrolyzing dextrin to glucose and estimating the glucose concentration by glucose kit = concentration of dextrins?

ii) Also, why dividing the glucose concentration by DP = actual concentration of dextrins? DP refers to degree of polymerization and I couldn't see how that division can give the dextrin concentration.

Thanks a lot!

Joseph

Because all dextrins are converted to glucose by hydrolysis by different enzymes

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC167871/pdf/621084.pdf

please go through --- http://www.ncbi.nlm.nih.gov/pmc/articles/PMC167871/pdf/621084.pdf

Sir i have studied some article related your question and finally found that my attached article will very help full and exact answer for your question. Thanking you

Thanks all. Let me digest all your inputs for a while. I have been doing computational work in the past, but now I need to do some experimental characterization and this is a fantastic forum to kickstart my work!

http://infoscience.epfl.ch/record/84293/files/91_Andriamboavonjy_JChromatogr.pdf

Hi,

Determination of your sugars by HPLC requires a standard to compare with. This is possible for linear (1-4 linked glucose), as there are standards available up to octaose (8 glucose units 1-4 linked). For longer sugars escpecially if also 1-6 linked it gets complicated. you could isolate (purify) these and detemine the concentration (see below).

As i dont know exactly what your needs are I suggest the following.

How you hydrolyze the starch? Enzyme, acids ? Thereby I could tell you what is ilkly to be produced and see how to quantify it.

In generall. Glucose, maltose and so on up to octaose can be quantified by HPLC easy enough

Glucose, can be quantified enzymatically (coupled to spectroscopy) See here http://www.pbc.ethz.ch/publications/index Hostettler 2011

The total amount of dextrins (or fractions of intrest coming from the HPLC) can be determined the same way just digest all your material to glucose by using a commercial glucosidase.

In theory you could to the same things (instead using the HPLC) by FACE (fluorophore-assisted carbohydrate electrophoresis) analysis. Which could be of advantage if you have larger prouducts which you might not see well by HPLC.

Ask me directly if something is not clear and I will provide you with more informations (references)

Sebastian

You can use a simple method to classify your product into three categories (mono. oligo and poly sacch.). Simply you can measure the amount of glucose by a suitable kit in three stages: 1) In your crude extract to measure the mono sacch. 2) after hydrolysis by HCl 0.05M for 30min at 100C to hydrolyze the oligo to mono, to measure the amount of glucose coming from the oligo, 3) after hydrolysis with HCl 1M for 4hr at 100C, to measure the amont of glucose coming from poly sacch. To calculate the DP you must know the MW of your poly sacch. firstly. This can be performed by using either HPLC-MS or gel filtration column.

I think separation and determination of dextrins can be done by RP-HPLC. :)

Make sure your HPLC equip with Reflective Index Detector, not UV or Fluorescence Detector. Good Luck.

Thank you guys... Is it possible to say using the HPLC to determine the peaks for glucose, maltose and maltotriose. After that, the remaining peaks which are not glucose, maltose nor maltotriose can be classified as "remaining dextrins"? Technically correct to do this? Please advise..

Sebastien Streb sounds like he's got it right; purchase quality standards for comparison of peak location to your unknown sample for identification of your dextrins. Known concentrations of the standards can then be compared to samples for quantitation.

I'm afraid, but I worked only about separation of protein in HPLC. I haven't experience about sugars..good luck!

For this purpose you need specific HPLC column used for separation of carbohydrates and RI detector or ECD detector. There are number of methods available for analysis of carbohydrates in Application section of column manufacturer. Method for analysis of dextrose is also available In different pharmacopeia.

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Dear Yong, be sure that dextrins and mono saccharides can not be separated by one column. The mono sacch. are usually separated by amino column, while dextrins are separated by gel filtartion column. The RP column is very weak to separate any of them. On other hand, if you used a mass detector (HPLC-MS) with both columns (amino and gel filtration) in two separat runs, you will get your target result. Good luck.

I'm afraid, but I worked only about separation of gangliosides in HPLC

Try to use Aminex HPX 87-P for detection of mono and oligosachharides. It worked for me.

Dear everyone,

Now I am doing the research related to analysing oligosaccharide during bioethanol fermentation. However I have some question related to storage the sample. I could store at -20C to inactivate every biological activity. By the way, during analysis with HPLC (auto sampler) the sample will be change due to activity of enzyme. Do you have any suggestion to analysis with high accuracy.

Thank you.