I prepared the standard solution for all sugars and got the specific RT for each sugar (Raffinose 7.8 and Sucrose 8.4). And I found Raffinose is high in leaves bcz its stress experiment but the problem in case of the sucrose sugar you can't identify the peak. I think this because of the high concentration of raffinose that make it not appear. Crop is Sugar Beet, Salinity stress experiment and sampling age is 30, 45 and 60 days
i suggest you to post the HPLC working condition that you are following,
like flow rate, mobile phase , column stationary phase etc...,
Therefore a proper modifying of procedure can be made in order to remove overlapping.
Method should be further developed. Your question needs further information.
i suggest you to post the HPLC working condition that you are following,
like flow rate, mobile phase , column stationary phase etc...,
Therefore a proper modifying of procedure can be made in order to remove overlapping.
Hi,
I think you have to play with the method you are implementing right now. As previous comments suggested, variations of the column, flow rate can help. Also modifing your gradient, increasing the run time may help to separate the components. Since the difference between these both sugars is a Gal unit you can use that info to play around. Maybe as you mentioned the amount of each sugar in the sample can be something that may affect results. Also are you sure under your experiment conditions (salinity stress) there are no variation on these compounds synthesis? I’m not really familiar with that topic, that is why I’m wondering. You may not see sucrose because somehow may be affected.
Hope you find some answers soon!
Annia mam I am very happy with your interest to answer me. You are right with that point about that raffinose synthesis is increasing under salinity stress, that's why it's overlapping with sucrose. But really I don't know how to solve it as you and my friend Omar mentioned in the previous comments because I am not sure if changing the time or flow rate will effect or not and really it's so costly to process and try many times without knowing about the results ☺️
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