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Plant Pigment Chromatography

Experiment #4A from Advanced Biology with Vernier

Education Level
High School


Paper chromatography is a technique used to separate substances in a mixture based on the movement of the different substances up a piece of paper by capillary action. Pigments extracted from plant cells contain a variety of molecules, such as chlorophylls, beta carotene, and xanthophyll, that can be separated using paper chromatography. A small sample of plant pigment placed on chromatography paper travels up the paper due to capillary action. Beta carotene is carried the furthest because it is highly soluble in the solvent and because it forms no hydrogen bonds with the chromatography paper fibers. Xanthophyll contains oxygen and does not travel quite as far with the solvent because it is less soluble than beta carotene and forms some hydrogen bonds with the paper. Chlorophylls are bound more tightly to the paper than the other two, so they travel the shortest distance.

The ratio of the distance moved by a pigment to the distance moved by the solvent is a constant, Rf. Each type of molecule has its own Rf value.

{R_f} = \frac{{{\text{distance traveled by pigment}}}}  {{{\text{distance traveled by solvent}}}}


In this experiment, you will

  • Separate plant pigments.
  • Calculate the Rf values of the pigments.


Teaching to an educational standard? This experiment supports the standards below.

International Baccalaureate (IB) 2025/Biology
C1.3.4—Separation and identification of photosynthetic pigments by chromatography
C1.3.10—Advantages of the structured array of different types of pigment molecules in a photosystem

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This experiment is #4A of Advanced Biology with Vernier. The experiment in the book includes student instructions as well as instructor information for set up, helpful hints, and sample graphs and data.

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