Chromatography columns need to be stored in appropriate storage solvents. If the filling material in the chromatography column dries out due to various reasons, it needs to be activated by low-flow wetting (i.e. activation) with the chromatography column storage solvent before normal use. Otherwise, at best, normal peaks cannot be obtained, and detection work cannot be performed. At worst, improper use may cause damage to the chromatography column. Therefore, if the filling material dries out, the chromatography column needs to be activated. So, what are the situations that can cause filling material to dry out and require activation of the chromatography column?
What situations can cause the packing material to dry out and require activation of the chromatography column?
New chromatographic columns may experience loosened end plugs during storage and transportation, leading to evaporation of the storage solution and subsequent drying of the column ends. In this case, the column needs to be activated according to the instructions, typically for 1-4 hours (refer to the instructions for specific guidance). However, if the peak shape is still abnormal after short-term activation, does that mean the column is damaged? Not necessarily, as this could be due to severe drying of the filling material that cannot be fully wetted by short-term activation. In this case, it is necessary to extend the activation time of the chromatographic column, typically by overnight activation at a flow rate of 0.2-0.3 mL/min using the activation reagent specified in the instructions.
Does an old chromatography column never need to be activated? No, for chromatography columns that have been used, if they have not been used for more than a month, the storage solution may also be lost, especially for chromatography columns that have not been removed from the instrument for more than a week, they need to be reactivated before use.
Does an old chromatography column never need to be activated? No, for chromatography columns that have been used, if they have not been used for more than a month, the storage solution may also be lost, especially for chromatography columns that have not been removed from the instrument for more than a week, they need to be reactivated before use.
What should be noted when activating chromatography columns?
● For chromatography columns that have been unused for a long time or are newly opened, it is generally not recommended to directly activate them using the detection flow rate. Instead, the flow rate should be maintained at a low level for a period of time before slowly increasing it. Otherwise, it is easy to damage the filler due to the sudden high flow rate, resulting in damage to the chromatography column.
● When taking chromatography columns out of the refrigerator or when the instrument is in a high-temperature state, it is important to allow the column to return to room temperature and the column temperature box to be lowered to room temperature before connecting the column. Otherwise, there may be a sudden temperature difference that could damage the chromatography column.
● If abnormal peak shapes are observed during detection and activation is required, it is important to first fully replace the solvents in the chromatography column with a transitional mobile phase before using the activation reagent. For reverse-phase chromatography columns, if the mobile phase contains salt, it is necessary to completely replace the salt with a low proportion of organic mobile phase before activation using the activation reagent.
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