1. Firstly, it should be confirmed whether the joints between the column and the instrument, as well as the pipelines, are compatible. To reduce dead volume, the inner diameter of the connecting pipeline between the injection valve, column, and detector should be as small as possible, while controlling the length of the connecting pipeline between the injector, chromatography column, and detector. Before installing the chromatography column, confirm whether the solvent in the flow path system is normal. It is recommended to install protective columns for samples with complex analysis.

2. In order to achieve optimal connection between the chromatographic column and the instrument system, it is recommended to use nuts and tapered joints that match the interface of the chromatographic column as much as possible. If the original joint matches other types of chromatographic columns, it is recommended to check the matching situation before connecting a new chromatographic column to avoid damage to the chromatographic column or leakage caused by mismatched chromatographic columns.

3. Universal joints made of PEEK material can be tightened by hand without the need for a specific wrench, with a pressure of 5000psi; The operating temperature should not exceed 100 ℃.

mobile phase

1. Before conducting sample testing, use at least 20 times the column volume of mobile phase to fully equilibrate the chromatographic column. The mobile phase must use solvents of chromatographic grade. If using aqueous buffer, it should be prepared on the same day to maintain freshness and avoid bacterial growth.

2. Before using the mobile phase, it is necessary to filter it with a microporous membrane to eliminate the damage of particles in the mobile phase to the chromatographic system and column. After mixing with other mobile phases, the buffer solution should be re filtered to avoid changes in solubility that may cause the formation of new precipitates. Pure water should not be used as the mobile phase to rinse C18 chromatography columns to avoid column performance damage (adding 5% organic solvent to rinse the chromatography column can also achieve buffer salt cleaning and make the chromatography column easier to equilibrate).

3. The mobile phase needs to be degassed before use to avoid abnormal operation of the pump and detector caused by bubbles. If there is a significant difference between the mobile phase used during testing and the mobile phase used for chromatographic column storage, an over distribution form should be used for equilibrium. Avoid damage to the chromatography column and instrument system caused by sudden changes in the mobile phase resulting in excessive increase in column pressure or crystallization of buffer salts in the mobile phase. A normal phase chromatography column requires a longer equilibrium time than a reverse phase chromatography column.

sample preparation

1. The sample should be dissolved as much as possible in a solvent that is miscible with the mobile phase. Unless otherwise specified, using strong solvents to dissolve sample columns may result in a decrease in resolution.

2. The sample solution should be pre filtered using a needle filter before injection. Frequent changes in the composition of the mobile phase will accelerate the reduction of column efficiency.

3. The chromatographic column is packed by high-pressure homogenization method and can withstand high pressure. To achieve separation efficiency, please do not exceed 200kgf when using, and avoid sudden pressure increases or changes, otherwise it will cause damage to the silica gel packing and reduce the service life of the chromatography column. Unless otherwise specified, the maximum operating temperature should not exceed 60 ℃.

Save operation

1. If the mobile phase contains acids or inorganic salts, it should be washed clean with deionized water (20 times the column volume) first. Then store the chromatography column in acetonitrile or methanol. Seal the joint of the pillar and store it in a stable environment.

2. The chromatographic column should be avoided from direct mechanical impact or falling to prevent a decrease in column performance.

Regeneration of chromatographic column

Rinse the column with a solvent equivalent to 20 times the volume of the column in the following order. It is recommended to rinse in the direction of the arrow on the column as much as possible to avoid backflushing. The reference solvent sequence used for flushing is water, methanol, chloroform, and isopropanol.