Dhruv Patel Yogesh Brahmbhatt August 30, 2010 Chromatography of Leaf Pigments Objective: Students will use chromatography

paper to separate plant pigments. They will then measure the Rf values of each pigment. Materials: 1. Chromatography paper 2. Long jar with cover 3. Ruler 4. Pencil 5. Quarter 6. Scissors 7. Pipette 8. Leaf (spinach) 9. Chromatography solvent 10. Waxed plastic wrap 11. Clip Procedure: 1. Materials were collected. 2. Chromatography paper was cut to the length of the jar. 3. About 1cm of the Chromatography solution was added to the jar. 4. Line was drawn 1.5 cm from the bottom. Leaf pigments were extracted by placing the leaf on the line and rolling the edge of the quarter. 5. Chromatography paper was placed in jar taking car that the green pigment line did not touch the solution. 6. The paper was clipped to the side of the jar. 7. The jar was then covered with the plastic wrap so that the solution would not evaporate. 8. The Chromatography paper was removed when the solvent was near the top. 9. The solvent line was marked with the pencil. 10. The paper was allowed to dry and then the bands were marked. 11. The measurements were made using the ruler. Data: Band number 1 2 3 92 13 10 Distance (mm) Band color Yellow orange Light yellow Bright green

Distance that the solvent moved: 106mm

Calculations (results): Pigment (color of pigment) Carotene (yellow orange) Xanthophylls (yellow) Chlorophyll A (bright green) Rf

Error: Error may have occurred due to the irregular spread of the original pigments on the bottom of the chromatography paper. Error may have also occurred if the pigments had touched the pool of solution at the bottom of the jar. Possible error may have occurred in measurement of the bands. Percent error calculations: % error of carotene: % error of xanthophylls: % error of Chlorophyll A: Conclusion: The pigments were separated in the experiment because different pigments have different affinity for the solution. The retention factor was calculated using the distance traveled by the band and the distance traveled by the solvent. This experiment reveals that there many other pigments beside the standard green in the leaf. Different leafs may have different amounts of different pigments and some may not have all of them.