Preparation of High Efficiency Columns for High Performance Liquid Chromatography

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Abstract
This method employs a specially constructed slurry reservoir, which features an internal taper and a quick-disconnect pump fitting. A major innovation is the use of a three-solvent system to maintain a slurry of packing material as a ‘slug’ within the reservoir. A measured amount of carbon tetrachloride is poured into the empty reservoir-column assembly. The column packing material is introduced as an isopropanol slurry and the reservoir is filled with isooctane. The assembly is then connected to a Haskel DSTV 122C pump, and the column is packed with isooctane at 10,000 psi pressure. This procedure provides HPLC columns with efficiencies of 40,000 plates per meter for 10-micron silica, 30,000 plates per meter for 10-micron C18, and 70,000 plates per meter for 5-micron C18 packing materials. We have used this method for preparation of over 35 HPLC columns during the past two years.