Separation of Chlorophyll a, Chlorophyll B, and Beta Carotene by Paper Chromatography

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Pigments extracted from different greens have different polarities and may be different colors. Mixed pigments can be separated using chromatography paper. Chromatography paper is able to separate mixed pigments due to their polarity and solubility. Pigments of chlorophyll a, chlorophyll b and beta carotene will be separated on chromatography paper because each has its own polarity and solubility, which results in different distance traveled up the paper. Beta carotene is non-polar so it travels the highest distance, followed by chlorophyll a. Chlorophyll b is the most polar; therefore, it travels the shortest distance. The separated pigments on the chromatography paper can be eluted in acetone and absorbance spectrum is
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Beta carotene is orange, and its graph created by spectrophotometer shows that it mostly absorbs blue and green light and reflects orange.

Figure 1. The absorption spectrum for leaf pigment, wavelength in nm.
During the separation of the pigments by chromatography paper, chlorophyll b traveled the shortest distance, chlorophyll a went above it, and the highest went beta carotene. This
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