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Photosynthesis and Chromatography of Spinach Leaves

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    Photosynthesis and Chromatography of Spinach Leaves Discussion: The chromatography of the spinach leaves was taken to establish what pigments were present in the chloroplasts of the leaves. The solvent carried and separated the different pigments up the cellulose paper. The solvent moved up the paper because of capillary action because of adhesion of the solvent molecules and the paper. The pigments dissolved in the solvent also were carried up the paper and were separated because they do not all dissolve equally into the solvent, less soluble pigments appear lower on the paper.

    The pigments also were attracted to the paper at different levels. Each pigment has a specific Rf value which is the distance the pigment migrated divided by the distance the solvent migrates. The data shows that Chlorophyll a has the highest Rf value, and it did travel the farthest out of the pigments. The control in this experiment was the same spinach leaves used. Error was no Carotene detected.

    Carotene is the very abundant in plants, so the lack of it found was based on reading errors of the chromatography, or the confusion of Xanthophyll with Carotene because the Rf value of Carotene is 0. 98, which was calculated for Xanthophyll. A spectrometer was used to measure photosynthesis by detecting how much light was able to pass through the cuvettes. The spectrometer was set to 100 percent transmittance of light in cuvette 1 because if photosynthesis occurred in the other cuvettes, they would transmit the same amount of light.

    DPIP was used to replace the final electron acceptor of photosynthesis, NADP. When DPIP accepts electrons from the processes of photosynthesis, it changes in color from blue to clear. The most light was able to pass through in cuvette 3 because it had unboiled chloroplasts that had light. The presence of light started the light dependent reactions and reduced the DPIP making it clear and allowing more light to pass through. The lowest percentage of light transmittance was in cuvette 4 with the boiled chloroplasts.

    The DPIP remained blue, and the boiled chloroplasts clumped together to further reflect light back instead of through. Controls in this experiment were the keeping of the cuvette 1 as 100% transmittance, the amount of chloroplasts used, the amount of DPIP, the amount of water, and phosphate buffer in the cuvettes. The error was in cuvette five, the values of the light transmittance should have been the same as cuvette 2. This error can be caused by faulty spectrometer readings, or fingerprints on the cuvette.

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    Photosynthesis and Chromatography of Spinach Leaves. (2017, Feb 14). Retrieved from

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    How do you extract photosynthetic pigments from spinach leaves?
    Extract photosynthetic pigments by grinding 2g of spinach leaves, torn into small pieces, in a mortar with a pinch of clean sand and a total of 10mL of 100% acetone. Initially, add only a small amount of acetone to begin the grinding process. It is much easier to grind the leaves if the extract is a pasty consistency.
    How does chromatography relate to photosynthesis?
    During photosynthesis, molecules referred to as pigments (due to the wavelength, thus color, they reflect) are used to capture light energy. Four primary pigments of green plants can easily be separated and identified using a technique called paper chromatography.
    What did chromatography reveal in the photosynthesis investigation?
    Chromatography and Photosynthesis Chromatography has revealed that many different pigments, not only green ones, are simultaneously present in leaves. Each pigment absorbs only certain colors of light from sunlight, rather than absorbing all the incident light energy that falls upon it.
    Which pigments in spinach leaves are visible during paper chromatography?
    The green chlorophyll breaks down in the leaves. As the bright green pigment fades away, yellow, red, and orange pigments become visible. Small amounts of these colors have been in the leaves all along.

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