Column chromatography is a separation technique that is used among many disciplines including biology, biochemistry, microbiology and medicine. Many common antibiotics are purified by column chromatography.1 Column chromatography allows us to separate and collect individual compounds. In this experiment, lumen will be the stationary phase, and the more polar substance will be retained on the stationary phase longer.
To start out the experiment, a short pipette which was used as the column was prepared. A small paper plug was placed loosely in the bottom of the column and then approximately ½cm of sand was added. After that, 6 cm of Alumina were added to the column, making sure that it settles evenly, and it was then topped off with another ½ cm of sand. The column was then attached to a ring stand using a clamp. Two watch glasses where then weighed and labeled. Approximately 2cm depth of Heptane to the top of the column was added. The entire F: AF solution was added immediately to the top of the column once the liquid level drained down and reached the top of the sand. When the F: AF solution dropped into the sand, heptane was added to the top of the column making sure it did not dry. The yellow band was collected in the first watch glass. Once all of the yellow band had eluted, the watch glasses were switched and when the heptane level reached the top of the sand, we switched to MTBE (methyl t-butyl ether). The MTBE was continued to be added to the column to elute the orange band. Both of the watch glasses were placed in a fume hood where the solvent was allowed to evaporate, afterwards, the watch glasses were weighed again. A developing chamber by outfitting a 100 mL beaker, and a watch glass for a lid was then prepared. Dveloping solvent was added to the chamber to a depth of no more than ½ cm. The mobile phase for ferrocene:acetylferrocene is 30:1 toluene :