MadSci Network: Chemistry
Query:

Re: How do you find a quantative analysis of glucose?

Date: Sun Oct 22 17:11:43 2000
Posted By: Kevin Ramsey, Staff, Speciality Sweeteners, Chr. Hansen, Inc.
Area of science: Chemistry
ID: 971277467.Ch
Message:

One of the most accurate methods of the determination of a carbohydrate is 
through the use of High Performance Liquid Chromatography.
High Performance Liquid Chromatography, HPLC, is an efficient way to 
separate macromolecules, ionic species, natural products, polymers, and 
other high-molecular weight groups of molecules.  HPLC achieves this by 
pumping a liquid mixture through a column at high pressure.  An 
interactive liquid mobile phase carries the mixture through the column, 
which contains a stationary phase. Components of the mixture interact 
differently with both the mobile and stationary phases as they are being 
pushed through the column at high pressure.  
Chromatographic separation in HPLC is the result of specific interactions 
between sample molecules with both the stationary and mobile phases so, 
components of a mixture elute from the column at different times.  
Separated fractions are collected by placing an open vessel at the end of 
the column.  Recovery is usually quantitative and separated sample 
components are readily isolated from the mobile-phase solvent.  A typical 
chromatograph will provide the amount of sample (eluent) vs. the retention 
time.  The area of a peak is proportional to the concentration of the 
component.  Retention time can be used as a qualitative analysis for 
describing an analyte because analytes elute through at different times.
Therefore you are able to detect qualatively the retention time of glucose 
by the use of a standard.  And quantitativly by the measurement of the 
peak area.


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