Operation and Tips for Management of HPLC Columns
The HPLC column uses the liquid as the mobile phase and the generalized solid phase as the stationary phase for sample separation. Therefore, the following points should be noted in operation:
It is advisable for all analytical samples to adjust the pH value to a suitable range for the HPLC column. Sampling overload should be avoided, which is one of the important methods to maintain the performance of the HPLC column for a long time.
If the manual injector is used for injection, the action should be fast and accurate. When the composition of the analyte is complex, the HPLC column should be protected by a protective column in front of the HPLC column.
At the beginning of each analytical work, the HPLC column should be balanced. The column and pipeline should be exhausted and flushed with an aqueous solution containing 10% organic solvent in the mobile phase.
The column should be flushed for about 30 minutes. The pure organic solvent in the column should be replaced. Then the column should be balanced by the mobile phase with at least 10 times the volume of the column until the baseline equilibrium is reached. Samples can be taken at the rear.
Operation of HPLC Columns
In daily separation analysis, it is significant to properly use a column or the service life of the column efficiency of it can be directly affected. In the course of column loading, unloading, or replacement, strong mechanical vibration should be prevented so as not to produce gaps in the column bed.
If the column is used for routine analysis, a special column may be arranged for each kind of conventional analysis to prolong the life of the column.
Additionally, avoid sharp changes in pressure and temperature, because the sudden temperature will affect the filling condition of the column and urge the packing in the column. And the rotation of the valve should not be too slow when the valve is injected.