If a student is purifying a compound using a reverse phase HPLC column, and the compound cannot bind to the column, what would be the possible solution to improve the interaction b... If a student is purifying a compound using a reverse phase HPLC column, and the compound cannot bind to the column, what would be the possible solution to improve the interaction between the column and compound?
Understand the Problem
The question is asking about potential solutions to improve the interaction between a compound and a reverse phase HPLC column when the compound cannot initially bind to the column. It explores options related to polarity and column characteristics.
Answer
Increase the polarity of the mobile phase during sample injection.
The final answer is to increase the polarity of the mobile phase during sample injection.
Answer for screen readers
The final answer is to increase the polarity of the mobile phase during sample injection.
More Information
In reverse-phase HPLC, a non-polar compound interacts more with non-polar stationary phases under less polar mobile phases. Adjusting mobile phase polarity affects solubility and retention.
Tips
A common mistake is confusing the polarity roles, thinking decreasing polarity helps. In reverse-phase, increasing mobile phase polarity can reduce compound solubility, promoting interaction with the column.
Sources
- Why is my protein not binding to my reversed-phase column? - researchgate.net
- Ionizable compound purification using reversed-phase flash column - biotage.com
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