I want to separate 2 compounds on HPLC, and I have made the preliminary studies on TLC using reversed phase plates and 100% ethanol as mobile phase which achieved complete separation. Now I want to apply this condition on HPLC. Ethanol high viscosity will increase the pressure or it will dissolve the column particles?
Yes you can but provided that to use HPLC grade ethanol. Despite that; if you are still in the development stage I prefer to start with methanol or acetonitrile as the organic modifier.
I think you most use another M. P. For seapration in Rp-HPLC like methanol :water (70:30). You Will get good seapration with no interactions between peaks.
The ethanol-water can be used as RP-HPLC mobile phase by increasing the column temperature moderately.
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