How to Activate and maintain A Common Used C18 HPLC Column?
The activation of a new HPLC column is actually a balanced process. In addition to using with a mobile phase to balance, it is sometimes especially important to balance the new HPLC column with the sample being measured, especially for peptides with higher molecular weights. Because of the high molecular weight of molecules, the diffusion rate is slow, the time required for the balance is correspondingly longer.
The specific balance way is very simple as well, just do more times for HPLC sample injection, and stop until the peak area and retention time is stable, and then the formal injection determination can be proceed.
If you want to speed up the equilibration time, you need to make the previously used to balance the injection sample concentration increased or continuous injection of multiple needles before unequal elution completed. The analyte is equilibrated to the new column in order to saturate the adsorptive capacity of the site on the silica matrix packing that has nonspecific adsorption.