ver.210629E
SHOWA DENKO K.K. (https://www.shodex.com/)
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7.4 Column Cleaning (Regeneration)
Problems in peak shapes and elution time changes or elevated column pressure etc. are often caused by
insoluble or adsorbing components present in the eluent and reagents being deposited inside the column. These
problems may be resolved by cleaning and/or regenerating the column. When washing and/or regenerate the
column, let the washing solution flow from the column outlet go directly into the waste container and not let the
solution go through the detector.
<Cleaning Method>
Insoluble components that block the column inlet may be removed by reversing the flow direction, i.e.,
introducing the eluent from the column outlet, with flow rate at less than half of the recommended flow rate.
<Regeneration Method>
Cations present in the sample may be adsorbed on the packing material. This can change the separation
patterns and peak shapes. Following regeneration procedure may resume the column performance.
Set the column temperature at 50 °C and the flow rate at 0.5 mL/min. Introduce 20 mL of 25-mM aqueous
sulfuric acid solution.
8. Column Storage
Remove the column from the system after replacing the in-column solvent with the shipping solvent. Securely
tighten the end caps and store the column at a location with stable temperature (a cool and dark space is
recommended). Refer to section 7.3 Solvent Exchange for how to replace the eluent.
Attention!
· Never allow inside of the column to dry. It can damage the column.
9. Column Inspection
Inspection method is described in the Certificate of Analysis (CoA).
Theoretical Plate Number (N) and Asymmetry Factor (Fas) were calculated using the below equations.
(1) Theoretical Plate Number (N)
(2) Asymmetry Factor (Fas)
