Exp. 14:  The Extraction of Chlorophyll from Spinach Using
Thin-Layer-Chromatography


Chlorophyll is the green coloring matter in plants and it plays an important in photosynthesis. Chloropyhll can be separated from other pigments in leaves such as those of spinach by the use of thin-layer-chromatography.

Procedure:

Part A:  Preparation of the spinach extract

Weigh out 0.5 g of spinach leaves and place in a mortar, Then weigh out 0.5 g of anhydrous MgSO4 and 1.0 g of sand. Add them to the mortar and then grind the spinach with the sand and magnesium sulfate for 5 - 10 minutes until a dry paste forms. Transfer the ground spinach to a small test tube to which you have previously added 2.0 mL of acetone.

Part B:  Preparation of the Chromatography Chamber

Add enough of the solvent mixture to cover the bottom of a 150 mL beaker. Cover with an inverted 250 mL beaker and leave for at least 10 minutes to allow the solvent and vapor to equlibrate.

Part C:  TLC Analysis

Cut a 3.5 cm x 9.0 cm TLC plate from the TLC sheet. Be careful to keep fingerprints off the dull side as they will interfere with the separation process of the spinach extract. Hold by the edges as much as possible. Use a pencil and draw a line across the narrow end of the plate 1 cm from the edge. Spot small portions of your sample onto the the TLC plate and then place the plate in the chromatography chamber and leave there until the solvent front has reached to within a couple of centimeters from the top of the plate. Remove from the chamber and let air-dry or dry with a dryer.


Information on setting up for thin-layer chromatographic separations

Results and Discussion:

Report the colors seen on the TLC plate. Also, (1) Measure the distance each band traveled from the original spotting point and (2) compare your results with those obtained by others in the class and

How is TLC used in the medical field and in forensic science?