System Condition: The analysis was performed on a Metrohm IC system. Separation of all standard mixture was achieved with the help of Hamilton RCX-30 column. An isocratic system was employed using sodium hydroxide (100 mM) solution in HPLC grade milli-Q water as a mobile phase including injection volume of 300 lL with a flow rate of 1 mL min-1. Different standards of sugars including Inositol, Rhamnose, Arabinose, Glucose, fructose, Sucrose and raffinose were analysed.
Problem: In total run time of 45min- Inositol(4.73min), Rhamnose(9.17min), Arabinose(10.78min), Glucose(12.13min), Fructose(13.41min), Sucrose(20.81min), Maltose(30.65min), and Raffinose(37.56min).
Apart from these in my plant sample there is another peak found at 5.58min just after Inositol. As there is no literature available regarding my plant so I'm having trouble in identifying that sugar. Please help me regarding this what standard sugar should I use to identify that peak?. What can be the possibility to identify that sugar?. Thank you
Check the application notes on sugars separation on your column or other similar columns from other vensors. Maybe you will find the answer on that chromatograms.
Regards
G
Glusoaminitol or fucitol are possible compounds which may elute shortly after inositol by application of NaOH.
Best wishes, Peter
Try to inject the mobile phase as a sample, if that peak appears so it is not a sugar. Another solution is that try to increase the flow rate to 1.5 mL min-1 and watch the number of peaks.
Dear Fathy the peak which I'm getting is a sugar because in some sample its area is low and in other samples its higher than other sugars. It is not the error of method or system. It is a sugar and I'm not able to identify that sugar. Thank you for your suggestion.
Dear Roos, thank you so much I will try Glusoaminitol or fucitol. Hope so it will help me.
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