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Pigment Chromatography Lab

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Introduction

Pigment Chromatography Lab Sahar Rudgar Mr. Wright SBI4U- Per. 4 Tuesday Mar. 22nd, 2005 Purpose: Separation of plant cell pigments by paper chromatography. Materials: * Chromatography paper * Scissors * Test tube * Chromatography tube and solvent * Pencil * Fresh leaf Procedure: 1.) A test tube was obtained containing developing fluid (9 volumes of petroleum ether / 1 vol. acetone) and a stopper with a hook. 2.) A 15 centimeter strip of Whatman chromatography strip was cut. 3.) One end to a point was trimmed with scissors. A pencil line was drawn just above the point. 4.) The strip on a hard clean flat surface was laid. A fresh leaf was put over the paper and the tissue was crushed onto the pencil line by rolling the edge of a test tube along the leaf surface. ...read more.

Middle

The distance that the solvent front moved was measured. The migration rate (Rf value) was calculated for each pigment. Observations: Pigment Colour Distance- Pigment Distance - Solvent Migration Rate (Rf Value) orange to yellow 6.0 cm 9.0 cm 0.667 yellow or grayish 4.0 cm 9.0 cm 0.444 green to blue-green 3.5 cm 9.0 cm 0.389 green to yellowish-green 1.5 cm 9.0 cm 0.167 Sample Calculations: Rf = distance pigment traveled distance solvent traveled = 3.5cm 9.0cm = 0.389 Discussion Questions: 1.) Chromatography separate molecules, because a sample containing two components is applied to a column containing a solid support coated with a given chemical layer (stationary phase); also present is solvent that is being applied continuously to the column (mobile phase); as sample components travel through the column, they interact with the mobile and stationary phases to different degrees; those interacting more strongly with the stationary phase stay in the column longer; results in separation of components as they pass through the column. ...read more.

Conclusion

It makes up about 25% of the pigmentation. 3.) Orange to Yellow: Beta-Carotene Yellow or Grayish: Xanthophylls Green to Blue-green: Chlorophyll a Green to Yellowish-green: Chlorophyll b 4.) In the fall, my sample leaf might turn yellow, red, or brown, because plants stop producing chlorophyll molecules due to lower temperatures. Error Analysis: Fingerprints on the filter paper may have affected the experiment because the oil from your hands can get on these things and affect the results. The spectrophotometer may have not been calibrated correctly because this was the first time this particular one had been used. Conclusion: The many pigments found in chloroplasts are all involved in gathering energy from sunlight. About four different pigments were present in the plant cell: Chlorophyll a, chlorophyll b, xanthophylls and beta-carotene. The spectrum of color displayed on the filter paper showed the pigments and the solubility of each. This indicated that photosynthesis was occurring and at what rate it was occurring. ...read more.

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