Cleaning Silica Gel Bonded HPLC Columns
The key to regenerating contaminated HPLC columns is to know the nature of the contaminants and to find the right solvent to remove. If contamination is caused by the accumulation of strongly retained materials during repeated injections, simple steps to remove these contaminants can often restore their chromatographic behavior.
Occasionally, after many operations, the column is rinsed with 90 to 100% solvent B (the stronger solvent in the dual solvent reversed phase system) to remove contaminants.
For example, the residual lipid in the column can be a nonaqueous solvent such as methanol, acetonitrile or tetrahydrofuran. If you are using a buffer system, do not switch directly to a strong solvent.
Sudden switching to a high concentration of organic solvent may cause buffering in the HPLC flow system, which can lead to larger problems such as plugging of the column, blockage of the pipe, pump Leakage, piston damage, or injection valve shaft failure.
The unbuffered mobile phase should be used first (i.e. the buffer should be replaced with water). Replace the strong solvent after rinsing 5 to 10 volumes.