Photosynthesis/ Chromatography Lab Photosynthesis- is a complex chemical process that converts radiant energy (light) to chemical energy (sugar).

Chloroplasts- They are bounded by a double membrane. The openings that enable CO2 to enter the leaf are stomata. Light must be absorbed before its energy can be used. A substance that absorbs light is a pigment. The primary photosynthetic pigments that absorb light for photosynthesis are chlorophyll a and b. Chlorophylls are not the only photosynthetic pigments, there are also accessory pigments such as carotenes and xanthophylls and they absorb light and transfer energy to chlorophyll a. Paper chromatography is a technique for separating dissolved compounds such as chlorophyll, carotene, and xanthophyll (method used to separate molecules from a mixture). Medium we used in class to separate the molecules was paper (paper is made from cellulose). When a solution of these pigments is applied to strips of paper, the pigments absorb onto the fibers of the paper. When the tip of the paper is immersed in a solvent, the solvent is absorbed and moves up through the paper. As the solvent moves through the spot of applied pigments, the pigments dissolve in the moving solvent. The pigment does not always keep up with moving solventsome pigments move almost as fast as the solvent, where others move more slowly. This differential movement of a pigments results from each pigments solubility and characteristic tendency to stick to the cellulose fibers of the paper. Pigments absorbed strongly move slowly, where those absorbed weakly move fastest. Hydrophobic molecules will move faster because they are more attracted to the hydrophobic solvent that the hydrophilic paper. On the other hand, hydrophilic molecules will move slower because they are attracted more to the paper than the hydrophobic solvent. You separate pigments from plants based on their solubility.

Chlorophyll- reflects green light. They are polar (water-soluble) pigments that act as electron-transporters in photosynthesis and provide green colors to plants. Chlorophyll a is more soluble than chlorophyll b and will travel further up the paper with the solvents being used.

Four Bands of Color Appeared on the Strip: 1) Carotene is located on the top of the strip and it moved the most with the solvent. It is the color orange (hydrophobic). 2) Xanthophyll is the second pigment on the strip and it is the color yellow. 3) Chlorophyll a is the third pigment on the strip and it is the color blue-green (since its close the bottom is it is hydrophilic). 4) Chlorophyll b- is the last pigment on the strip located all the way on the bottom and it is the color yellow-green (hydrophilic). Rf: The ratio of the distance traveled by a particular substance to the distance traveled by the solvent, both measured from the original spot, constitutes the so-called ratio of fronts, or Rf, for that substance. This value is constant for a given set of conditions, but changes when the conditions are changed, for example if a different solvent is used. Lab Questions 1. Which is more soluble in the chromatography solvent, xanthophylls or chlorophyll a? xanthophyll 2. If yellow xanthophylls were present in the extract, why did the extract appear green? They look green because theres so much chlorophyll that masks the other pigments. 3. What does a small Rf tell you about the characteristics of moving molecules? The smaller Rf value, the heavier the molecule and the more soluble. 4. What color of light was most effective for plant photosynthesis? Violet, blue and green 5. What color of light was least effective for plant photosynthesis? Yellow and green.