Reversed-Phase Partition Chromatography

Abstract

In the course of experiments involving the use of powdered glass as a carrier in partition chromatography, we have found that it is possible to prepare columns having as the stationary phase either an aqueous liquid or a water-immiscible organic solvent. Columns in which a stationary aqueous phase is distributed on glass have already been applied to the fractionation of the alkaloids of Punica Granatum¹. We now find that satisfactory reversed-phase columns may be prepared by the following procedure. Chloroform (8 c.c.) is shaken thoroughly with ‘Pyrex’ glass (80 gm.) in No. 100 powder. The ‘wetted’ glass is tipped into water (100 c.c), previously saturated with chloroform, and the mixture is mechanically stirred at moderate speed with an upflow stirrer until all globules of chloroform are disintegrated; the slurry is poured into the chromatogram tube and again thoroughly shaken. The column is allowed to pack spontaneously by draining in a vertical position, and the top is tamped by gentle pressure on a disk of filter-paper placed above the packed column. Similar columns can be prepared using benzene instead of chloroform. Attempts to prepare this type of column employing kieselguhr as carrier were unsuccessful.