Extracting Pigments from Spinach

The leaves of green vegetables contain chlorophyll, which can be extracted using the following method.


Materials Required

• Spinach leaves
• Mortar & pestle
• Pasteur pipette (dropper)
• Test-tube
• Acetone
• Hexane

Procedures

1. Tear the spinach leaves into small pieces and place them in a mortar.
2. Add approximately 10 mL of acetone and carefully grind the sample and acetone together with the pestle. Continue grinding until the acetone becomes dark in colour. More acetone may be added to compensate for evaporation.
3. Using a disposable Pasteur pipette, extract the acetone and place it in a small test tube. Be careful not to extract the small pieces of vegetable.
4. Repeat steps 2 and 3 using the same vegetable remaining in the mortar, until the test tube is approximately 1/3 full.
5. Add approximately 3 mL of hexane to the acetone extract.
6. Mix the contents of the test tube by using a clean disposable Pasteur pipet to repeatedly withdraw a sample and then immediately dispensing the sample back into the test tube. Be careful not to let the test tube overflow.
7. Let the mixture stand to allow the two layers to separate.
8. Draw out the upper hexane layer from the test tube.

This dark green extract can be separated on Thin Layer Chromatography:

See a video demonstration by Peter Keusch (scroll down the page to the end of "experimental procedure").