HPLC Column-operating Guides

The following operating guides are recommended for maintaining reversed-phase columns. Consult the vendor s column instructions for precautions and column regeneration guides. 

Store the columns in acetonitrile or methanol or in a mixture of water and organic solvents. Cap them with closed fittings to prevent the columns from drying out. 

Do not exceed the pH range of the column (typically pH of 2.5 to 8 for silica-based columns). Silica dissolves at pH greater than 8 and bonded groups can hydrolyze at pH less than 2. Note that many modern silica-based columns may have more durable bonding chemistries for extended pH usable range of 2-10 or even 1-12.10  

Do not exceed the temperature limits of most columns, usually 60-80 C, without consulting the instructions from the column vendor. 

Always flush the column with a strong solvent (methanol) before use to eliminate any highly retained analyte. 

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The following operating guides are recommended for connecting HPLC columns ... 

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