How components will seperate in hplc column & what do you mean by polarity?



Answer:
Through the interaction with the resin. If the resin is hydrophobic and you try to separate hydrophobic compounds, the most hydrophobic compound will interact with the resin the strongest so it will be retained on the column the longest; the opposite will be true for a less hydrophobic or polar compound. Also, separation may depend on the size and surface of the resin - small compounds may be retained longer than large ones, because they will partition between the solid phase (the resin) and the mobile phase (the eluent) (size exclusion chromatography). Finally, you may have ionic interactions between the resin and the compounds you want to separate, primarily in ion-exchange chromatography.

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