Hello Everyone,
Please let me know, if there is any single or different methods available for detection of glucose, xylose and xylitol by HPLC equipped with C18 column and UV detector. If not please let me know the best methods for detection of the above with HPLC in detail (conditions like run time, mobile phase, columnan and its conditions and detector or suggest an alternative columns for HPX 87H.
Thank you
Try using a carbohydrate ES column from Prevail™ to detect dextrose, Xylose and Xylitol. Use a gradient method for this analysis.
Mobile Phase Solvents: A: Acetonitrile B: Water.
Flow rate: 1.0 - 2.0 mL/min
Detector: ELSD
Hello Golla : in what media do you want to detect your samples ? Are you isolated from natural sources or other sources .
With your equipment the best I can see is trying to separate xylose from glucose with only distilled water as mobile phase at 1ml/min or less with UV detection at 190nm.
Using a NH2 phase column and a RI detetor will be a more convenient, or onther carbohydrate coolumn.
Column HPX-87P, with water (HPLC grade) as eluent,0.6 mL/min, RI detector, sample: 20 microliters
These sugars mentioned above can not be detected by UV for their are no UV absorption groups.
You can either try derivation method - thus can still use C18 column and UV detector, or change to NH2 column and use ELSD or RI detector.
Agree with Chunhua Yang, you will spend a lot of energy trying and money but you will never have something correct with that equipement.
Uv detector is not selective for monosaharides, for that detector you will need to do derivatisation of analityts and then analyze it on HPLC.
Column: Aminex HPX 87 H
Mobile phase: 5 mM Sulfuric acid @ 0.6 min/ml
Detector : RI
Temperature: 45 C
See the attached file for actual chromatogram
Dear Golla, you can find procedure in detail in the National Renewable Energy Laboratory method "Determination of Sugars, Byproducts, and Degradation Products in Liquid Fraction Process Samples".
http://www.nrel.gov/docs/gen/fy08/42623.pdf
Best regards
Frantisek
Is there any other method to find xylose concentrations other than HPLC
Dear Golla, you can use either Aminex HPX-87P (water as mobile phase) or some NH2 column (acetonitrile/water mobile phase). Detection is possible via RI or ELSD detector. Carbohydrates can be analysed also by GC. Details are in attached files.
Hi dear
In addition to Aminex HPX-87P, the shadoex sugar Sp0810 can be used with sulfuric acid 0.005 M as a mobile phase and flow rate is 0.6 ml/min and column temp. is 55- 60 c and run time 50 min
good luck
Amino-silica column with acetonitril:water (60:40), 0.6ml/min flow rate
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