To separate the chloroplast pigments by paper chromatography. Concentrate the extracted chlorophyll solution by evaporation. Apply a drop of it at one end, 2cm away from edge of a strip of chromatography paper and allow it to dry thoroughly. Take a mixture of petroleum ether and acetone in the ratio of 9:1 at temperature of 40 to 60°C. Hang the strip in the jar with its loaded end dipping in the solvent. Close the jar tightly and keep it for an hour. The pigments separate into distinct green and yellow bands of chlorophyll and carotenoid respectively.

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Pigments are the molecules which reflects only certain wavelengths of visible light. Chromatography is the technique used to separate the chloroplast pigments. Carotenes form yellow-orange band, chlorophyll forms blue-green band, chlorophyll b forms yellow green bands.

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