Lab Report Chromatography

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.

The concentrations and absorbances of the red and blue dyes were used to find the concentration of the purple dyes. From the graph of the blue dye, the linear equation for absorbance was y = mx + b. From that formula came the equation y = 7.915 x 104 (x) + 0.02489, where y represents absorbance, m is slope, x is concentration/molarity, and b is the constant/y-intercept. The same set up was performed for the red dye, but the equation produced was y = 1.045 x 104 (x) +.001298. The equations found when graphing absorbance vs. concentration were used to find the concentration of the purple dyes. The absorbance for purple dye 3 on the red wavelength of 470 nm equaled 0.149 and 0.818 for the blue wavelength of 635 nm. For purple dye 1

If a sample’s Rf is lower, that means that it remained closer to the stationary phase. In both variations of the experiment, red dye #40 remained the closest to the stationary phase (on average). Therefore, the red dye #40 had the lowest rate of flow value. The intermolecular forces of the red dye are responsible for its low distance traveled. The molecules of the chromatography paper have strong intermolecular bonds that are highly polar. The red dye is attracted to the paper because of their similar characteristics. The negative and positive ends of both the paper molecules and the red dye #40 molecules attract to each other. It is extremely difficult for the isopropyl alcohol and the sodium chloride solutions to interfere with the bonds of paper and ready dye #40, as a result, both solutions are unable to fully dissolve red dye #40. . NaCl and isopropyl alcohol have very strong intermolecular forces, but the forces of the paper’s molecule were able to attract red dye #40 because they were even

If you put too much of a sample on your chromatography paper, you could possibly have that color bleed into the color next to it, which would mess up your results. If you put too little of a sample on the paper, your color

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.

The dyes in the laboratory experiment are made of numerous colors, mainly red and blue, the spectra from each of the dyes corresponded to the wavelengths obtained from each of dye i.e. 620 nm for red and 450 nm for blue.

3. Now you need to extract the dye from the solution onto the strips of yarn. To do this, place one piece of yarn into each test tube of colored solution. Heat the tubes in a boiling water bath for 8-10 minutes. You can remove the tubes from the water when the solution is milky-white and the yarn is the color of the dye, in other words, when all of the dye has been extracted from the solution.

Chromatography is a fairly simple process. First, you put a dot of ink(or in our case, the M&M food dye) near the bottom of some chromotography paper (also known as filter paper), and then hang the paper vertically with its lower edge (the one closest to the spot of dye) dipped in a solvent (In our case, the sodium chloride solution). Capillary action forces the solvent to travel up the paper, where it meets and dissolves the ink. The dissolved ink (which is the mobile phase) slowly travels up the paper (the stationary phase) and separates out into its different elements. Another way of describing it is to think of the liquid as an adhesive-like liquids, some of which stick more to the solid and can travel more slowly than others. This is

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.

This experiment demonstrated the separation of pigments based on relative polarity and proved to be a substantial way to separate compounds. The results were much like that of an experiment performed, which separated carbohydrates in a very similar method with the use of paper chromatography (Inome, Y., & Yamamoto, A.). Proper pipetting technique, which is described by John Husler, was also demonstrated in this experiment. The technique was followed as to prevent contamination and deliver the right amount of solution each time (John Husler: 1983).

The solvent moves through the stationary phase by capillary action (it is attracted to the thin layer of water in the cellulose) and picks up the molecules that are attracted to it. To determine the level of separation of the mixture, Rf values are used. Rf values are given by the ratio of the distance the dye travels divided by the distance the solvent travels. The farther the dye travels, the more attracted it is to the solvent and the greater the Rf value. Water is the best solvent to separate the dyes because it is the most polar of all the solvents and it is attracted to ions (forming an ion dipole moment.)

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

The first step of the procedure was to take a large 600 mL beaker and pour 10 mL of the mobile phase (NaCl in H2O solution) into it. Immediately following, the beaker had been covered with aluminum foil instead of Parafilm. Next, we took the chromatography filter paper and drew a horizontal line with hash marks to represent where the dyes would be spotted. The paper and hash marks were labeled properly. Only pencil could be used because the ink from other utensils would have dispersed throughout the paper when it touched the mobile phase. In order to make the spots, we had to make a well plate of the different dyes. Once that had been completed my lab partner and I spotted our chromatography paper using broken toothpicks as our dotting tool. We let each spot dry before adding more dye to keep its small size. A new toothpick had been used for each individual dye to prevent mixing of the dyes. After each spot dried the chromatography was stapled without the edges touching and placed into the 600mL beaker restraining from coming into contact with the sides of the beaker. Now we had let the paper sit to absorb the solvent and as it nears the top, the paper was carefully removed. Then, the paper had been placed into the oven to dry. Afterwards my partner measured the appropriate distances to calculate the Rf values

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

Gel-Filtration Chromatography is a commonly used method used in order purify a protein from a mixture, by means of separations. Different biomolecules differ in size, or their molecular weight. In the gel matrix inside the chromatography column, there are gel beads which are porous to allow certain sized molecules to enter. The molecules that are able to enter the pores of the gel, are held in stationary phase and will elute from the column later on, these are usually smaller, to medium sized molecules. Larger molecules that are not able to fit in the pores will elute out of the column first, they are involved in mobile phase where they just go straight through the column without interacting with the gel beads. Smaller molecules will have a higher elution volume, while the larger molecules will have a lower elution volume. The volume to elute the protein is inversely proportional to the molecules size.