How do you regenerate a C18 column?

How to regenerate the C18/C8 HPLC columns?

1. Disconnect the column from the detector.
2. Rinse with a mobile phase without buffer salts (water/organic phase) firstly.
3. Then rinse with 100% organic phase (methanol and acetonitrile).
4. Check that the pressure returns to normal, and if not, proceed to the next step.

What is regeneration of column chromatography?

Column regeneration. Exposure of a column to samples or solvents containing highly adsorptive components will result in increased back- pressure and a change in selectivity. Often the column can be restored to original performance by suitable wash protocols.

How do you clean C18 columns?

Wash the column slowly over to 100% methanol and wash for at least 15 minutes. Wash the column over to 100% acetonitrile and wash for at least 15 minutes. Depending on the type of column you are using you can store the column in 100% organic depending on the manufacturer’s recommendations.

Is C18 reverse phase?

A C18 column is an example of a “reverse phase” column. Reverse phase columns are often used with more polar solvents such as water, methanol or acetonitrile. The stationary phase is a nonpolar hydrocarbon, whereas the mobile phase is a polar liquid.

How do you regenerate a silica column?

Regeneration Procedure for Normal-Phase Silica Column

1. Disconnect the column and reconnect it to the system, with the flow through the column in the reversed direction.
2. Set the flow at 1 mL/min.
3. Flush column with 50 mL of ethyl acetate.
4. Reconnect the column in the proper flow direction.

Is C18 gel polar or nonpolar?

In other terms, C18 has Octadecyl chains which are usually hydrophobic and highly retain nonpolar compounds. The length of its carbon chain is longer. On the contrary, C8 has Octyl chains and hence it is less retained when used with the same compound as C18.