Column and Thin Layer Chromatography: the Separation of Spinach Pigments
Title Page
Chelsea Maksin
Organic Chemistry Laboratory 1
Column and Thin Layer Chromatography: The Separation of Spinach Pigments
Russ Hoburg
02/15/2012
Abstract
The main goal in doing the column and thin layer chromatography was to separate spinach extract into its components based on polarity and then to analyze the components. The separation of the spinach extract was done using the column chromatography with the wet/slurry packing method. Alumina was inserted into the column to act as the stationary phase. After the components were separated and the solvents boiled out, the thin layer chromatography was performed to analyze the components of the spinach extract. This was done using a TLC plate and spotting each of the components on to the plate as well as spotting the standard spinach extract. A beaker was used with the developing solvent, filter paper, and TLC plate inside and a watch glass over top of the beaker to perform the thin layer chromatography. After the solvent front (30% ethyl acetate in hexane) reached about an inch from the top, the TLC plate was removed from the beaker and the developing spots were marked. After implementing the column and thin layer chromatography, the Rf value for component one was .95, for component two it was .28 and .61, and for the third component it was .41. The first component was the same as one of the components of the standard but the final two components were slightly off from the components of the standard likely due to human error.
Introduction
Chromatography is the technique of separating components of a mixture based on its physical properties. The separation is done between two phases known as the stationary phase and the mobile phase and depends on if one component is more likely to adhere to the stationary phase more than another component. Moreover, the stationary phase is almost always more polar than the mobile phase [Padias pg. 163]. The rate at which these components move through the column
Chelsea Maksin
Organic Chemistry Laboratory 1
Column and Thin Layer Chromatography: The Separation of Spinach Pigments
Russ Hoburg
02/15/2012
Abstract
The main goal in doing the column and thin layer chromatography was to separate spinach extract into its components based on polarity and then to analyze the components. The separation of the spinach extract was done using the column chromatography with the wet/slurry packing method. Alumina was inserted into the column to act as the stationary phase. After the components were separated and the solvents boiled out, the thin layer chromatography was performed to analyze the components of the spinach extract. This was done using a TLC plate and spotting each of the components on to the plate as well as spotting the standard spinach extract. A beaker was used with the developing solvent, filter paper, and TLC plate inside and a watch glass over top of the beaker to perform the thin layer chromatography. After the solvent front (30% ethyl acetate in hexane) reached about an inch from the top, the TLC plate was removed from the beaker and the developing spots were marked. After implementing the column and thin layer chromatography, the Rf value for component one was .95, for component two it was .28 and .61, and for the third component it was .41. The first component was the same as one of the components of the standard but the final two components were slightly off from the components of the standard likely due to human error.
Introduction
Chromatography is the technique of separating components of a mixture based on its physical properties. The separation is done between two phases known as the stationary phase and the mobile phase and depends on if one component is more likely to adhere to the stationary phase more than another component. Moreover, the stationary phase is almost always more polar than the mobile phase [Padias pg. 163]. The rate at which these components move through the column
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