Thin Layer Chromatography Essay

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Analytical Applications of Thin Layer Chromatography Linh Nguyen Santa Barbara City College Abstract. For the purpose of this lab, thin layer chromatography (TLC) is used to analyze and identify the twenty different amino acids that are commonly used in the synthesis of proteins. Based on the data that is obtained from the experiment, we have concluded that the molecular compounds’ polarity can determine their migration rates over the TLC plate. As a result, while hydrophobic molecules have strong affinities to the nonpolar solvent and thus have large Rf values, hydrophilic molecules are more attracted to the polar silica gel and will therefore have low Rf values. Introduction THIN-LAYER CHROMATOGRAPHY, OR TLC, is a scientific method…show more content…
First, starting 1 centimeter from the end of a plate, a pencil is used to lightly draw a line that is 1 centimeter from the lower line of a TLC sheet. With a capillary pipette, each of these amino acid solutions is spotted evenly along the origin line. While that is being dried, a developer with a 4:1:1 ratio of butanol, citric acid, and water is poured into a glass developing tank that is covered with a lid. Then, a piece of paper towel that is submerged in the developing solution will be adhered to the inside wall of the developing tank. After the pH of the developer is measured and recorded, the TLC sheet is placed upright in the glass container so that the edge with the spots is slightly above the solvent and the aluminum backing is not leaning against the saturated paper towel. When the solvent finally reaches approximately 8 centimeters above the origin line with the help of capillary action, the TLC plate is dried thoroughly and submerged in a solution that contains ninhydrin. After it is blow-dried, the TLC sheet is placed in the oven so that better visualizations of the amino acids can be made. We can measure the distance from the origin to the center of each spot (X) as well as the distance from the origin to the solvent front (Y). The division of X by Y will allow us to obtain the Rf value for each of the amino acids (Doohan,

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