Preservation and Regeneration of the HPLC Column

1.The preservation of the HPLC column

(1) Maintenance of the reversed-phase column after the daily experiment:
The buffer or saline mobile phase was used and rinsed with 10% methanol/water for 30 minutes after the experiment was completed. The salt in the column was washed off and rinsed with methanol for 30 minutes.

(2) long-term preservation of the HPLC column:
If the column is to be stored for a long time, it must be stored in a suitable solvent. For the reverse phase column, it can be stored in pure methanol or acetonitrile. The normal phase column can be stored in pure n-hexane after dehydration. The ion exchange column can be stored in water (containing preservative sodium azide or thimerosal). Plug the plug that came with the purchase of the new column. The stored temperature is preferably room temperature.

2. Regeneration of the HPLC column

Because the HPLC column is a consumable, as the time of use or the number of injections increases, the peak height decreases, the peak width increases, or the shoulder peak appears. Generally, the efficiency may be reduced.

(1) Regeneration of the reversed-phase HPLC column: sequentially using 20-30 times the column volume of methanol: water = 10:90 (V/V), acetonitrile, isopropanol as the mobile phase to wash the column, and then Rinse the column in the reverse order. Such as

(2) Regeneration of normal phase HPLC column: The column is washed with n-hexane, isopropanol, dichloromethane, and methanol as the mobile phase in the order of 20-30 times column, and then the column is washed in the reverse order. It is to be noted that the above solvents must be strictly dehydrated.