Separation of Photosynthetic Pigments by Paper Chromatography

Separation of Photosynthetic Dyes by Paper Chromatography Introduction Chlorophyll is actually only one of a closely related group of dyes called photosynthetic pigments commonly found in photosynthetic plants. This can be demonstrated by extracting pigments from the leaves with acetone and separating them by paper chromatography. Fortunately, five pigments can be distinguished: chlorophyll a (blue - green), chlorophyll b (yellow - green), lutein (yellow), carotene (orange) and fucoid (this is the decomposition product of chlorophyll It is a thing.

* Understand the chromatographic separation process and separate the photosynthetic pigment mixture extracted from the leaves. * Separation of dye and leaves by paper chromatography. * Identify the mixture by dividing the mixture into different compounds. * Compare the pigments on various leaves to see if there are similarities and differences. Materials used in this experiment were microcapillaries, 100 mL Erlenmeyer flasks and weighing, 3 paper chromatograms, and mortar and pestle. The specimens used are Zea mays, gumamela (Hibiscus rosa-sinensis) and San Francisco (Codiaeum variegatum). There are five steps to extract and separate pigments. The first step was to place 0 grams of the new grounded blade of the above sample in 5 to 10 milliliters of acetone in a test tube. Insert the tube, shake intermittently and place it on the tube for about 10 minutes.

The purpose of this experiment was to isolate the various photosynthetic pigments found in the leaves of spinach and to extract them using paper chromatography. The aim is also to determine the relative amounts of chlorophyll a and chlorophyll b from their absorption spectra. Photosynthetic pigments were separated from fresh spinach leaves and their absorption and percentages to chlorophyll a and chlorophyll b were measured. In addition to paper chromatography, another method called spectrophotometry is used to measure the light absorption of each pigment. The retention (Rf) value of each pigment and the concentration of chlorophyll a and b were calculated and all the results are tabulated. Next, the ratio of chlorophyll a to chlorophyll b was calculated, 6: 1, which is very close to the expected result.

Chromatography is a technique for distinguishing several dyes in plants. Photosynthetic pigment of spinach is chlorophyll b and carotenoid. However, theoretically, it should contain four pigments, chlorophyll a, chlorophyll b, lutein and carotene. During the chromatography process, select a mixed solvent that separates the individual pigments during the analysis, place the loaded chromatogram in a solvent and move the solvent upward, and if the front side of the solvent comes close to the paper, analyze Stop.