Rf Value Lab Report

The spinach plants were hole punched, mixed with 0.2% NaOH and dish detergent, placed in a syringe with the solution to have the oxygen

The purpose of this experiment is to determine the maximum absorbance of fast green, and the chlorophylls, also in the case of fast green create a concentration curve to determine an unknown substance. Each test will use the spectrophotometer.

The procedures for experiment A, B, and C all start the same. The first step is to put on goggles and get the data collection device set properly. The labquest needs to be plugged into the colorimeter accurately so that a click is heard when putting it in. The labquest needs to be reading digitally and the colorimeter needs to be set to 635 nm. Then shake the chloroplast solution and take a clean cuvette and fill it with 3 mL of distilled water, 3 drops of the chloroplast solution, and cap it. This is used as a blank to calibrate your labquest. Double check that the labquest is reading absorbance, this assures that the colorimeter is plugged into the labquest accurately. Insert the blank into the colorimeter and hit the calibration button. Take out the blank and empty it. The labquest is now set to experiment with. Make sure that the heat bank is set in front of the lamp and that the lamp is on. The cuvette must be placed on the opposite side of the heat bank in the path of light in the box so that no other light can interfere with the experiment.

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.

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.

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.

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

The procedure to conduct the experiment, the effect of various powers of microwave radiation on the growth of grass plants is fairly simple. First, to perform this experiment, one must gather the necessary materials. These include 250 grass seeds, 25 paper cups, 3675 mL of soil, approximately 50 paper towels, microwave oven capable of being used at 100 Hz, 500 Hz and 900 Hz, one measuring cup(to measure materials), one liter of water, masking tape, sharpie marker, environment with ample sunlight and air, a ruler with centimeter markings, stove capable of heating up to 34°C, metal pot capable of holding up to 100 mL, thermometer, and a stopwatch (stopwatch application on a device will also work). Next, lay out the paper towels under the work environment. After that, place the

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

In the experiment conducted, paper was used as the stationary phase, and chromatography solvent was used as the mobile phase. With the help of chromatography paper and chromatography, three types of pigments were found in the plant.

The results should also be similar or slightly higher than the data collected for the red light. Results in step three showed a correspondence between the portions of the leaf that contained chlorophyll and those that contained starch (the brown colored portion of the “after” leaf sketch in Image 2.). The correspondence between these two sections of the leaf is that where there is chlorophyll, there is light being absorbed. There were portions of the leaf in which did not have any starch nor chlorophyll present. This was due to the variegation of the leaf; where there is no chlorophyll on a leaf, there is no reaction that can take place meaning that photosynthesis will not occur in that region and the starch will not be present in that area because of the lack of chlorophyll.

For lab 12, it is hypothesized that chlorophylls a and b are present in a plant leaf and contribute to the starch production in photosynthesis. Also, products of photosynthesis will be present in leaf tissue exposed to red and blue light wavelengths for several days, but a decreased presence in leaf tissue exposed to green and black light wavelengths. In lab 13, it is expected that since chlorophylls a and b are more polar and smaller molecules than the anthyocyanins and carotenoids, they will travel higher up the chromatography paper than the other pigments.

The following procedure dealt with a chromatogram. The materials needed are: a pencil, safety goggles, scissors, chromatography paper strip, capillary tube, spinach plant pigment extract, test tube, cork stopper, graduated cylinder, chromatography solvent (alternative isopropyl alcohol), metric ruler, stopwatch or clock with a secondhand, hook/fashioned paperclip, paper towels, test tube rack, and mortar and pestle. First we obtained a strip of chromatography paper and cut it so it would fit inside a test tube (with it barely touching the bottom of the tube). Also, when touching the strip, touch the sides only. Then we attached (firmly) the top of the strip to a hook (or fashioned paperclip at bottom of the cork stopper). Make sure it fits in the test tube. Next we used the pencil to draw a faint line across the strip two centimeters from the bottom tip of the strip. We placed the cork and strip in place, and we put a mark on the test tube one centimeter below the top of the stopper.

Chromatography Investigation Chromatography is a highly regarded technique used to separate the components of a mixture. It is based on the principle that each component possesses a unique affinity for a stationary phase and a mobile phase. The components that are more inclined to enter the mobile phase will migrate further on the chromatogram and distinguish themselves from the other components. The type of solvent used in chromatography is known to directly affect the separation of the mixture. In this experiment, thin-layer and column chromatography will be utilized to separate the numerous chlorophyll and carotenoid pigments of a spinach extract.