Thin Layer Chromatography
Thin layer chromatography is a method used to determine the purity of a solution or monitor a reaction progress. A TLC sheet is comprised of a silica gel. Based on the polarity of the molecule and the solvent used, each molecule will move a different distance on the gel. After the solvent moves the entire length of the sheet, the spots from the each molecule can be viewed under UV light.
A rotovap is a device used for the efficient and gentle removal of solvents by evaporation. A sample is attached to a vacuum while rotating in a heated water bath. The solvent then evaporates at a lower temperature than usual and recondenses on the cold finger which is filled with dry ice.
Column Chromatography is a way to purify a mixture of compounds based on the polarity of the individual molecules. The glass tube is filled with a silica gel that allows less polar molecules to travel through the column more quickly. Multiple fractions can be collected and then TLC analysis can be used to determine whether each fractions is completely pure.
H NMR is a method to determine whether we made the correct compound. Each hydrogen atom in the compound shows up as a different peak. Each peak can be compared to literature values.
Part of our research this summer was to determine whether the compounds we made were inhibitors of the enzyme beta-galactosidase using a standard procedure to test the rate of an enzyme. We added the enzyme to a substrate solution and, after a certain period of time, the solution turned yellow and the absorbance was able to measured. From there, we could determine the rate of the enzyme.
Members of Jameson’s Research lab for the Summer 2016: