Extraction and Isolation of Plant Pigments: Flow Chart Ekta Patel Exercise A Materials: Gloves Chromatography Paper Spinach Leaf Coin Pencil Chromatography Jars with Solvent Ruler 1. Measure 1.5 cm from the bottom of the Chromatography paper and make a pencil line across the paper (wear gloves). 2. Put a clean spinach leaf on top of the drawn line. 3. Place a ruler on top of the line and using it as a guide roll the edge of the coin over the leaf. 4. Repeat this process again by using different points of the leaf to create a dark pigment extract line. 5. Pick up and unscrew the chromatography jar underneath the hood. 6. Fold the chromatography paper in thirds and place it in the jar with the extract line just above the solvent line. Cap the jar. 7. Keep checking the paper until the solvent is about 1 cm from the top. Remove the paper and recap the jar. Allow the paper to dry. 8. Once the paper is dried determine the Rf values for each pigment: carotenes, xanthophyll, chlorophyll a, and chlorophyll b. 9. Clean up with the following procedures: Gloves in orange labeled trashcan, spinach in green labeled trashcan, return jar to fume hood, pour solvent into waste jar, return materials used, and wipe down station with Windex.

Prepare Chromatography Paper

Submerge Paper in Solution Remove Paper from Solution Calculate Rf Values

Exercise B Materials: Spectrophotometer scissors Chromatogram 2 cuvettes Kimwipes Calculators Color pencils acetone 1. Cut out the four different pigment sections from the paper from Exercise A. Place pieces in labeled bowls. 1. Allows the pigments to elute from the paper strips into the bowl by stirring the acetone with a glass rod. 2. Obtain the absorption values for the four different pigments with the following: a. pour the acetone into a cuvette b. fill another cuvette with pure acetone c. determine the absorption from 380nm-740nm. 3. Record this data in lab manual data table. 4. Clean up with the following procedures: dispose chromatography paper and gloves in orange labeled trashcan, wipe off table, empty cuvettes in waste container, rinse supplies and return to table, and wipe down work bench.

Prepare paper for acetone

Absorption Spectrum Plot data