Leaf Pigment Chromatography

Leaf pigment chromatography is a classic paper chromatography experiment that allows students to separate and observe the different pigments found in plant leaves. In this leaf pigment chromatography lab, students use chromatography paper and a solvent to see how pigments such as chlorophyll and carotenoids move at different rates, creating distinct bands of color. This chromatography experiment provides a visual and hands-on way for students to connect molecular properties to key biological concepts, including photosynthesis and plant structure.

This paper chromatography lab is especially effective in high school biology classes because it reinforces both content knowledge and essential lab skills. As students analyze the separation of leaf pigments, they practice careful observation, data analysis, and experimental design while gaining a deeper understanding of how plants capture and use light energy. Leaf pigment chromatography is a low-prep, high-impact lab that fits naturally into units on photosynthesis, plant biology, and scientific investigation. 

Check out this blog post on Paper Chromatography for additional photos and information.

Leaf Pigment Chromatography Experiment

Here are photos that I took as my students were working on their lab.

Spinach leaves are placed in a mortar and pestle along with a little acetone.  Students grind the leaves until the acetone turns a dark green.  A small piece of capillary tubing is used to transfer drops of the pigment extract to a piece of chromatography paper.

When enough drops have been placed on the chromatography paper to make a dark green circle on the paper, it is ready to be placed into a large 25x200 test tube.

Leaf Pigment Chromatography Diagram

The large test tube has about 1/2 inch of petroleum ether in the bottom of the tube.  The pigments will immediately begin to separate.

Here is the finished chromatogram:

Pigment Chromatography FAQ's:

What pigments can be seen in leaf chromatography?
Plants contain multiple pigments such as chlorophyll, carotenoids, and sometimes anthocyanins. These pigments separate based on their chemical properties when the solvent moves up the paper. 

Why do pigments appear in different bands?
Pigments travel different distances during chromatography due to differences in solubility and how strongly they bind to the chromatography paper. 

Can this experiment predict fall leaf colors?
Leaf chromatography can reveal hidden pigments, which often correspond to colors visible during autumn. 

Here is the lab that I use with my students:

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