Validation of washing procedures for maintaining a microbiologically clean gel filtration column.
Chromatographic gel filtration matrixes used in various separation techniques are subject to microbial contamination. The need for a microbe-free column is critical when preparing materials that require a low or zero microbial count. This report proposes two alternative washing systems: 0.02 N HCl containing 0.81% NaCl, and 0.1 M tromethamine--hydrochloride buffer (pH 7.0) containing 0.81% NaCl and 0.02% thimerosal. These washing systems were validated using a 100 x 2.6-cm column packed with a modified dextran gel slurry previously inoculated with known counts of USP test organisms. After each wash, the column separation characteristics were verified further with appropriate test proteins.
D001419:Bacteria; D001431:Bacteriological Techniques; D002850:Chromatography, Gel / Q000295:instrumentation / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D008628:Mercury / Q000032:analysis; D003904:Mitosporic Fungi; D013171:Spores, Bacterial; D013172:Spores, Fungal; D013849:Thimerosal; D014325:Tromethamine
D014325:Tromethamine; D013849:Thimerosal; D008628:Mercury