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Maintenance of HPLC Columns

1. Try to protect the HPLC column with a pre-column or guard column;
2. The pH range of most HPLC columns is 2-8, try not to exceed the range;
3. Avoid drastic changes in the composition or proportion of the mobile phase;
4. The mobile phase must be filtered and degassed before use;
5. If a polar or particulate buffer is used as the mobile phase, the column should be rinsed clean after the experiment.

C18 columns cleaning regeneration

Wash with the following 10 column volumes of solvent: (flow rate during forward flush: 0.5 ml/min)
95% water: 5% acetonitrile (or methanol)
(removal of buffer salts)
If the system does not have a gradient function, add 5 times column volume to 50% water: 50% acetonitrile (or methanol) rinse step.
95% acetonitrile (or methanol): 5% water
Finally stored in 35% water: 65% acetonitrile (or methanol).

Regeneration of reversed-phase columns

Sequentially use 20-30 times the column volume of methanol: water = 10:90 (V / V), nitrile, isopropanol as a mobile phase to wash the column, and then rinse the column in reverse order.

2018-12-10T05:25:15+00:00 December 10th, 2018|