Two gradients to use for analysis of hydrophilic and or hydrophobic molecules in the same Aqueous Normal Phase ANP method - Tips & Suggestions
April 14, 2020
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Some of the usefulness of a Cogent TYPE-C™ column is that you have options in selecting the gradient for a mixture of compounds that have either or both polar and non-polar compounds in it.
It might be best to try both a Reversed-Phase gradient (from 80% to 20% aqueous) and then an ANP gradient (from 20% to 80% aqueous) as an initial screening process. From the preliminary results you can determine which method is most likely to produce the best separation.
Once the direction has been selected the gradient can be modified to reach the desired retention and separation of the components in your mixture. Even an isocratic method can be learned from this quick and easy exercise.
It might be best to try both a Reversed-Phase gradient (from 80% to 20% aqueous) and then an ANP gradient (from 20% to 80% aqueous) as an initial screening process. From the preliminary results you can determine which method is most likely to produce the best separation.
Once the direction has been selected the gradient can be modified to reach the desired retention and separation of the components in your mixture. Even an isocratic method can be learned from this quick and easy exercise.