How to Use HPLC Column Correctly?
Sample preparation and operation:
1. The sample should be dissolved as much as possible in a solvent that is compatible with the mobile phase. Unless otherwise specified, the resolution of a sample column using a strong solvent may be reduced.
2. The sample solution should be pre-filtered with a syringe filter before injection. Frequent changes in the composition of the mobile phase will accelerate the reduction of efficiency.
3. The HPLC column is filled by high-pressure homogenization method and can withstand higher pressure. For best separation, please do not exceed 200kgf when using, and avoid sudden rise or change of pressure, otherwise, it will cause damage to the silica filler and reduce the service life of the liquid chromatography column. The maximum operating temperature should not exceed 60 °C unless otherwise specified.
Save the HPLC column:
1. If the mobile phase contains acid or inorganic salts, rinse it first with deionized water (20 column volumes). The column was then stored in 100% acetonitrile or methanol. Finally, seal it with the joint of the column and store it in a stable environment.
2. The HPLC column should be protected from direct mechanical impact or fall, to avoid the performance of the liquid chromatography column.
Regeneration of HPLC column:
Rinse the column in the following order with a solvent equivalent to 20 column volumes. It is recommended to rinse in the direction of the arrow of the column, as far as possible without backflushing: the reference solvent used in the rinse is water, methanol, chloroform, isopropanol, methanol.