I want to further fractionate my water fraction for isolating active molecule.
HP-20 is a polymeric reverse phase, so it will behave pretty much like C18 in terms of a separation mechanism. Start with water, and run a gradient with methanol or ethanol as described above. One can use acids (trifluoro, formic, or acetic acid) if needed to sharpen peaks.
For loading the sample, sounds like your sample is soluble in water so a liquid load is appropriate.
Pack your column in water and elute with water followed by ethanol or methanol.
HP-20 is a polymeric reverse phase, so it will behave pretty much like C18 in terms of a separation mechanism. Start with water, and run a gradient with methanol or ethanol as described above. One can use acids (trifluoro, formic, or acetic acid) if needed to sharpen peaks.
For loading the sample, sounds like your sample is soluble in water so a liquid load is appropriate.
Can we use simple column which we use for silica gel column chromatography for Packing?
Thank you very much sir. One more question is there if you can help then it would be great. Can we fractionate all type of molecules ( non polar to highly polar) with HP20? or it is only be useful for polar compounds?
I think go for polar molecules with HP-20. Use silica(60-120) for separation of non-polar molecules.
Yes, I've often run CHP20 in open glass columns. You may want a little air pressure to help keep the flow rate up.
Generally, yes, it can be reused. Wash with methanol or acetonitrile, then water. As this is a polymeric reverse phase (styrene divinyl benzene) I would _not_ recommend using solvents like dichloromethane to wash it since it may swell. it will eventually become discolored, but still work for some time.
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