How to Use and Maintain HPLC Columns?

Generally speaking, the reversed-phase high-performance liquid chromatography column cannot be backflushed. Only when the producer indicates that the column can be backflushed, the impurities remaining in the column header can be backflushed. Otherwise, the recoil will quickly reduce the efficiency of the HPLC columns.

Choose to use a suitable mobile phase to avoid damage to the stationary phase. Sometimes a pre-column can be connected in front of the injector. When the analytical column is bonded to silica gel, the pre-column is silica gel, so that the mobile phase can be “saturated” by the silica gel before entering the analytical column to avoid dissolution of the silica matrix in the analytical column.

Avoid directly injecting samples with complex matrices, especially biological samples, into the HPLC column. Pretreatment of the sample is required or a guard column is connected between the injector and the column. The guard column is typically a short column filled with a similar stationary phase. The guard column can and should be replaced frequently.

After the separation and analysis work is completed, it should not be stopped immediately. It is necessary to rinse the chromatographic analysis system in time, generally, 0.5h or more, to remove impurities in the HPLC columns.