SNAP
®
SeriesGlass Columns
Cleaning Instructions for PackedColumns (CIP)
Cleaning a SNAP
®
Column involves three Stages:
1. Regeneration of the column packing
2. Sterilization
3. Depyrogenation
Regeneration removes chemical and organic contamination that binds to the chromatography material, which significantly
reduces the capacity and resolution of the column. Contamination is usually caused by lipids and pyrogens, protein aggre-
gates, pigments, polyphenols andmetal complexes.
Sterilization removes and/or denatures microorganisms and spores with chemical treatment, otherwise they could contami-
nate the purified product. Ethanol solutions containing sodium hydroxide or acetic acid are the most commonmethods for
sterilization.
Depyrogenation involves the breaking-down of endotoxins that have become attached to the chromatography material or
the column hardware. (frits, tubing, etc.) Endotoxins can soil the target compounds in question by being washed gradually
through the column. Somemethods used to sterilize equipment will break down pyrogens.
Sterilize and Purify the columnwith the following steps:
1. Disassemble the column into individual parts
2. Wash in a dilute solution of caustic soda or sodium hypochloride (leave frits in solution for 30-60minutes)
3. Wash parts in a sterile, pyrogen-free solution and reassemble
4. Pack column in sterile environment
5. Solvents and solutionsmust also be sterile and pyrogen-free. Essential Life Solutions recommends in-line filtration
through a 0.22 μm filter.
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