Paper Chromatography Term paper
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Paper Chromatography Introduction: Paper chromatography is used to separate and identify pigments and other molecules from cell extracts that contain complex mixtures of molecules. In 1903 Michael Twsett invented this technique. In 1944 Martin and Synge, who went on to win the Nobel Prize in 1954 for their work, perfected it. Paper chromatography was used by Melvin Calvin to separate the order of photosynthesis, and Fred Zanger to find out how to make insulin by determining what orders to put the amino acids in. Trial 1 (Using paper chromatography to separate colors in a marker or dye): Hypothesis The colors in the black marker or dye will separate into different bands across the chromatography paper. The green dye will not have as many separate colors as the black marker. Procedure 1. Obtain a piece of chromatography paper and make a pencil line about 1.5cm from the bottom of the paper. Draw a line with the marker (or dye) over the pencil line.2. Staple the paper into a roll so that it will stand by itself.3. Put about 1cm of water into the bottom of a glass container.4. Put the rolled paper, marker line side down, into the glass container so that it does not touch the sides of the glass container and the marker does not touch the water.5. Wait until the water has moved to about 1cm from the top of the paper and remove the paper from the container.6. While the paper is wet, and the colors are still bright, draw a pencil line separating each color.7. Measure how far each color traveled to determine Rf values. Results Band Number Distance (mm) Band Color Rf Values1. 5 Yellow .112. 10 Orange/Yellow .213. 23 Orange .494. 34 Pink .725. 38 Light Pink .816. 47 Blue/Black 1 Trial 2 (Using paper chromatography to separate pigments in a spinach leaf): Hypothesis The colored pigments in the spinach leaf will separate in colored bands across the paper similar to the way the colors in the black marker separated. The trial using the homemade extract will show the separation of pigments better than the trial where the spinach leaf is rubbed onto the paper using a quarter. Procedure (When using the homemade extract) 1. Make the extract:a. Obtain 1g of spinach leaves and grind them into a paste using a mortar and pestle.b. Transfer the paste into a test tube.c. Add 4ml of Acetone to the mortar and pestle to rinse it out and then pour it into the test tube.d. Cork the test tube and shake it hard for 10sec.e. Let the test tube and mixture set for 10min.f. Add 4ml of water to the test tube and shake for 10sec.g. Add 4ml of petroleum ether to the test tube and shake for 10sec.h. Allow the test tube and mixture to set until a thin line forms of the top of the mixture. This is the extract.2. Obtain a piece of chromatography paper. Draw a line in pencil about 1.5cm from the bottom of the paper.3. Wash the paper strip by wetting it from bottom to top in petroleum ether:acetone solvent (1 part acetone, 9 parts petroleum ether) in a petri dish. Set the paper aside to dry.
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