Chromatography lab
Purpose: To separate food colorings into their component dyes using paper chromatography.
Materials: Chromatography paper, Food coloring, Ruler, Pencil, Solvent solution, Test tubes, Test tube rack.
Safety precaution: wear aprons, to make sure that you don’t get any of the alcohol on your clothes, and if you break a test tube you don’t get glass on you.
Procedure: See-attached handout.
Results: See chromatography with Audrey’s lab report.
Rf values for approved FD & C dyes
Dye Rf Value Dye Rf Value
Red No. 2 0.81 Yellow No. 6 0.77
Red No. 3 0.61 Green No. 3 1.00
Red No. 4 0.67 Blue No. 1 1.00
Yellow No. 5 0.75 Blue No. 2 0.79
Rf values for
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You would expect to find that component near the origin, because it was attracted enough to the developing solvent to move up the chromatography paper.
Discussion/Conclusion:
In this chromatography lab, we separated food coloring into their component dyes using chromatography paper. When we put the piece of chromatography paper in the solvent, the paper absorbs that liquid due to capillary action. The basic colors of the ink spot will separate out into bands, and you can see the different colors that are in the ink to produce that specific color. The smallest molecules will travel basically at the same rate the solvent travels up the chromatography paper. The larger molecules will travel slowly and will stay near the bottom. Our results of our experiment were a little different then the other groups. We all matched only a couple of the Rf values, but some of our distances were a little different. We probably took our chromatography paper out before or a little later then some people, because not all of our colors were had the same measurement. It would be interesting to try using a different kind of solvent to see how much of a difference it makes when the colors separate, because our blue never really separated into the different colors that make it. It would be cool to see either if you need to use a difference solvent or leave the chromatography paper in the solvent longer then we
1. Record your hypothesis about what will happen when Biuret solution is mixed with the solutions from test tubes 1, 2, 3, and 4 here. Be sure to use scientific reasoning to support your hypothesis.
7. Tape the strip to a pencil and rest the pencil on top of the jar so that the strip hangs into the jar. The goal is to have the end of the chromatography strip just touching the surface of the solvent solution, with the colored dots above the surface of the liquid. Make sure that the colored spots do not come in direct contact with the liquid in the bottom of the glass.
For part one of this experiment, I only experienced separation of colors with the green and brown M&M’s, along with the yellow food coloring. The green M&M separated into yellow and blue, with blue travelling farther up the paper. It is not surprising that green separated into blue and yellow because those are the primary colors that make up green. The brown M&M separated into red and orange, with the orange travelling farther up the paper. Finally, the yellow food coloring separated into yellow and red, with yellow travelling farther up the paper. This could be because it was such a concentrated, small amount of food coloring. The colors that didn’t travel very far up the paper, such as orange and brown, are probably less soluble than the others, like blue and green.
Answer: Once the chromatogram has been completed and is ready to be measured and calculated, on the plate that was used to perform the chromatogram you should see where the red and blue have completely separated. The red food coloring dye should be lower on the plate than the blue food coloring dye.
Abstract: The purpose of this lab is to separate and identify pigments and other molecules within plant cells by a process called chromatography. We will also be measuring the rate of photosynthesis in isolated chloroplasts. Beta carotene, the most abundant carotene in plants, is carried along near the solvent front because it is very soluble in the solvent being used and because it forms no hydrogen bonds with cellulose. Xanthophyll is found further from the solvent font because it is less soluble in the solvent and has been slowed down by hydrogen bonding to the cellulose. Chlorophylls contain oxygen and nitrogen and are bound more tightly to the paper than the other pigments.
In the first part, there may have been cross contamination of materials, especially since periods before us had already performed the experiment. During chromatography, our spot sizes were initially very varied, as we didn’t know exactly how much of each substance to place when we began. Thus, the substances in more abundance would have greater opportunity to travel up with the eluent, skewing the results. In the beginning, we even put 6-7 drops, causing some of the substances to mix together. Touching the TLC plate may also have disrupted some of the substances traveling up.
1. Paper Chromatography is a method used for the separation of colors which are also referred to as colored chemicals/substances or pigments. This method is used for experiments, to identify coloring agents and to separate out a compound into its various components.
Thin Layer Chromatography (TLC) works in relation to the polarity of chemicals. A plate is first covered with aluminum foil or silica etc. and has solutions of varying polarities placed upon the aluminum foil or silica. When placed in a in a puddle of solvent that moves up the plate, the different inks i.e. the solution will move up the place based on their Rf values. Adherence increases with increasing polarity, so the less polar compounds will be carried farther by the solvent. Eventually the dyes will separate into their compontnets, which can be visibly seen. This is then used to determine who the ink of the unsigned note belonged to along with the pen that it belonged
On a thin chromatography plate, five spots were placed ( as shown in table 2) and the plate was developed using chloroform/methanol. This was later visualized with dragendorff’s reagent under the UV light. All separated components were observed, identified and recorded.
Pigments extracted from different greens have different polarities and may be different colors. Mixed pigments can be separated using chromatography paper. Chromatography paper is able to separate mixed pigments due to their polarity and solubility. Pigments of chlorophyll a, chlorophyll b and beta carotene will be separated on chromatography paper because each has its own polarity and solubility, which results in different distance traveled up the paper. Beta carotene is non-polar so it travels the highest distance, followed by chlorophyll a. Chlorophyll b is the most polar; therefore, it travels the shortest distance. The separated pigments on the chromatography paper can be eluted in acetone and absorbance spectrum is
Chromatography is a separation technique in which the mixture to be separated is dissolved in a solvent and the resulting solution, often called the mobile phase, is then passed through or over another material, the stationary phase. The separation of the original mixture depends on how strongly each component is attracted to the stationary phase. Substances that are attracted strongly to the stationary phase will be retarded and not move alone with the mobile phase. Weakly attracted substances will move more rapidly with the mobile phase.
After wearing the gloves we obtained a chromatography vial from professor and label it with my and my peer initials. We dried up the chromatography vial in fume hood and added 1 ml of chromatography solvent to the vial. Then we took a chromatography strip and measure it 1.5 cm with ruler from one end of the strip and drew a line with pencil we cut two small pieces below the pencil line to form a pointed end. We applied spinach on the strip using quarter to rub the spinach leaf on the line that we drew on the strip and put it into the chromatography vial and placed that in fume hood. We observed as the solvent was moving up the chromatography strip by capillary action. When the solvent was reached approximately 1 cm from the top of the strip then we removed the cap from the vial. We took out the strip from the vial using forceps and marked up the location of the solvent front because it evaporates quickly. We measure out the distance as well as the pigment in order to find out the rf value. Moreover we compared rf values to the one in reference list in order to identify the
It is possible to separate these pigments from each other using a technique called paper chromatography. In this process, plant tissue extract is applied to a piece of chromatography paper. “A solvent is allowed to travel up the paper, and if the pigment is soluble in the solvent, it will be carried along with it.” (Benya, 2009) Different pigments have different affinities for the solvents or polarity and will travel at different rates. Chlorophyll, anthyocyanins, and carotenoids are typically non-polar.
Remove the tubes and add 2-3 drops of Iodine – potassium – iodide solution to each tube.
The next step was to place the strip of chromatography paper on a paper towel. Then dip a capillary tube into the plant pigment extract (spinach pigment extract) provided by the teacher. The tube will fill on its own. We applied the extract to the pencil line on the paper, blew the strip dry, and repeated it three to four times until the line on the paper is a dark