Can anyone suggest a method for determination of sugars (glucose, fructose, maltose, sucrose, mannose) by using RP-HPLC?
For Sugar analysis,
You have to use ACN/Water(78:22), flow rate=1ml/min, ELSD detector. But you have to use NH2 column. Try with C-18. A little optimisation is required with this column.
Gudluck
:)
Thanks....i will try .... I use ca ion exchange column , water as mobile phase , ..... But it need long time washing so not applicable to large no of samples is there's any recommidation to that column
Dear try this combination, i am sure it wl work.....
Solvent system; 5 mM H2SO4, Flow rate 0.5 ml, Column Te
mperature 30 C, Colum Specification Aminex -
HPX-87H Ion Exclusion Coulmn (300 mm*7.8 mm), Detector RID...
Please feel free to inquire other details..
Best of luck...
Pl use latest technology column is HILIC column. Pl find the attached file as application note.
Yes, Hydrophilic-Interaction Chromatography is latest and advanced technology as said by Dr Gupta. Go with this...
go with Muhammad Younus ..i ll advice you the same ...you can play with the retention time by varying the temperature( max 60 degree C) and by adding the organic modifier ( ACN in different proportion 10-60% ) ...conc of H2SO4 can be varied from 5mM to 8mM.
you can also go with Ajai gupta .............but before purchasing the column ,compare the prize of both or other available columns for carbohydrate separation.
It is not necessary that you only buy from a particular vender as I mentioned earlier. Whenever any new technology comes any where in the world. Other manufacturer also starts making the same with some improvements in less price. If need about the venders pl reply.
Do not use HPX-87H to determine sucrose, the H2SO inverts it. Use a HILIC column if you have no Protien of ogliosacc in you samples, if you do use a Calcium resin (Lead for more separation) which is Ligand exchange. or use IC. if you want cheap use FSCE (sometime called CZE), even better.
If you are determined to use RP. here is a good paper: "Liquid chromatographic profiling of monosaccharide concentrations in complex cell-culture media and fermentation broths"
you have used the HPX-87H or HPX-87p to quantify arabinose and mannose?
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