Cleaning of liquid chromatography column

1、 Introduction to the cleaning of liquid chromatography column:
Up to now, reversed phase chromatography is the most widely used technique in high performance liquid chromatography (HPLC), mainly because it is suitable for the analysis of most non-polar substances and many ionizable and ionizable compounds. When the reversed-phase method is used, the possible matrix interaction must be considered. When the chromatographic column is polluted, its chromatographic behavior will be slightly different from that of the unpolluted column The column will have back pressure problem.
The contaminated RP column must be cleaned and rebuilt to restore the original operating conditions. Usually, the sample contains contents that the analyst is not interested in. Salts, lipids, lipid containing substances, humic acids, hydrophobic proteins and other biological substances are substances that can interact with the HPLC column in the process of use, and the retention value of these substances is less than or equal to that of the analyst Objective.
2、 Materials and reagents required for cleaning of HPLC column:
Methanol, chloroethane, tetrahydrofuran
3、 Cleaning steps of liquid chromatography column
The key to regenerate the contaminated HPLC column is to understand the nature of the pollutants and find a suitable solvent to remove them. If contamination is caused by the accumulation of strong retentive substances during repeated injections, the chromatographic behavior can usually be restored by removing these pollutants with simple steps. Sometimes, after several operations, the column is flushed with 90% to 100% solvent B (the stronger solvent in a dual solvent reverse phase system) to flush 20 volumes to remove contaminants.
For example, the remaining lipids in the column can be used with non-aqueous solvents such as methanol, chloroethane and tetrahydrofuran.
4、 Precautions for cleaning liquid chromatography column:
If a buffer system is used, do not switch directly to a strong solvent. A sudden switch to a high concentration of organic solvent may precipitate the buffer in the HPLC flow system, which may cause more problems, such as column head blockage, pipeline blockage and pump blockage, leakage, piston damage or injection valve shaft failure. The buffer free mobile phase should be used first (i.e. changing the buffer to water), After washing 5 to 10 volumes, replace the strong solvent.
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