Separation And Separation Of Spinach Pigments By Paper Chromatography

Have you ever really wondered how different variables can affect how plants go through photosynthesis? Well, in this experiment, the purpose was to see how various environmental conditions can affect the overall photosynthetic capacity of a specific plant. The factors, light, darkness, cold, and heat were applied to see how the different components would affect the photosynthesis on spinach plants. Each group was given a different factor to test. Out group was given the light factor. The hypothesis for this experiment is that when adding light as a factor, the light will affect the overall plant photosynthesis.

There are two main types of chlorophyll, chlorophyll a which absorbs wavelengths of 430nm (blue) and 662 (red) and is the main photosynthetic pigment, and chlorophyll b, which doesn’t directly participate in the photosynthetic process, but is capable of donating its energy to chlorophyll a

Photosynthesis occurs each time the sun’s light reaches the lives of a plant. The chemical ingrediants for photosynthesis are carbon dioxide (CO2), a gas that passes from the air into a plant via tiny pores, and water (H20), which absorbed from the soil by the plant’s roots. Inside leaf cells, tiny structures called chloroplasts use light energy to rearrange the atoms of the ingrediants to produce sugars, most importantly glucose (C6H12O6) and other organic molecules. Chlorophyll gives the plant its green color (Simon, 02/2012, pp. 92-93). Chemical reactions transfers the sun’s light energy into the chemical bonds that hold energy-carrying molecules. The most common are

The image above is our chromatogram, you can see at the top the orange bands of a carotene, next are the bands of xanthophylls which are yellow. The next light green bands directly bellow the yellow is chlorophyll a, the darker green bands are chlorophyll b. In order to test absorbance of the chlorophylls they were cut out of this chromatogram submerged into acetone and then used to get an absorption spectrum as seen below.

Photosynthesis is the conversion of light energy to chemical energy into sugars. It is the process in plants that uses carbon dioxide, water, and sunlight from its surroundings and releases oxygen as a byproduct (6H2O+6CO2+light energy -> C6H12O6+6O2). Photosynthesis is required for plants because they are autotrophs, organisms that make their own food. Plants require a specific environment that is ideal to them to be able to carry out the process. Environmental conditions can either increase or decrease the rate of photosynthesis. Things like colors of light, pH, and temperature can all affect the rate of photosynthesis in plants.

Photosynthesis is the process in which plants consumed inorganic materials like solar light, carbon dioxide and water and converted it to an organic molecule like sugar and an inorganic gas like oxygen. Light is one of the major elements influencing the rate of photosynthesis; direct light concentration affects the noncyclic pathway (light

In the beginning of this experiment, our TA added water, salt, and 75/25 hexane/acetone to spinach leaves to a blender and blended the mixture to assume equal amounts for each group and to avoid erros if each student had to do the blending. The addition of the water to the mixture allowed the it to separate into a distinct organic layer after being run in a centrifuge, which was available to be collected at the top of the centrifuge. Salt reduces solubility, which will force the organic parts of the mixture (the desired pigments for example) to separate into the organic layer at the top. Lastly, 75/25 hexane/acetone is added because this is a moderately polar solvent and will useful for both the non-polar and polar pigments present within the spinach leaves. A mixed solution of hexanes and acetone must be used because acetone is very polar, while hexane in very non-polar, and the spinach leaves contain both non-polar and polar pigments in them that are important in the extraction and for further analysis. The mixture was placed in the centrifuge so the solids in the mixture (mostly cellulose) could be separated from the liquids into separate distinct layers for further extraction and testing. In the tube, the organic substances separated into the top layer, whereas the water layer remains at the bottom of the tube below the solid layer made up of mainly cellulose.

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.

During the separation of the pigments by chromatography paper, chlorophyll b traveled the shortest distance, chlorophyll a went above it, and the highest went beta carotene. This

If you expose a photosynthesizing plant to water that contains both radioactive H and radioactive O, in which of the products of photosynthesis will the radioactive H and O show up?

The purpose of this experiment was to take spinach leaves and extract the chlorophyll and carotenoid pigments by using acetone as the solvent. The chlorophyll and carotenoid pigments were extracted by using column chromography and alumina was used as the solvent. Solvents of different polarities were used, starting with the least polar, to extract the certain components from the leaves. They were then analyzed by using thin- layer chromatography.

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.

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

Without photosynthesis we would not be able to receive energy. We should be more appreciate of plants, without them we would not survive. This paper will explain the basic components require for photosynthesis, the role of chlorophyll, how energy is transferred, and photosystems I and II and the most precious product results of photosynthesis.