1. BSC 197: Molecular and Cellular Basis of Life Lab

2. Chromatography Mobile Phase- petroleum ether + Acetone
Hydrophobic/nonpolar
“Like dissolves like”
Nonpolar solvents dissolve nonpolar molecules and carry them up the chromatography paper
Solid Phase- chromatography paper
Hydrophillic/polar
Paper = polar material, polar molecules will stick to it and not migrate far up the paper

3. Look at the pigment structure on pg. 50 of lab manual
You can use the structure identify which molecules will be polar or nonpolar
Hypothesize which molecule will travel the furthest and why

4. Chromatography Lab
NO SMOKING! During down time and for at least 30 mins after lab
Goal: To use Rf values to identify the pigments present in carrots and spinach
In order to do this, you need to have controls:
½ the class will do lutein, chlorophyll B, carrot, and spinach
½ the class will do beta-carotene, chlorophyll A, carrot, and spinach

5. Control data Experimental data Carrot Spinach Beta-carotene Lutein
Chlorophyll A
Chlorophyll B
Experimental data
Carrot
Spinach

6. Divide the filter paper into 4 parts by folding it into quarters
Each group will draw a line 1 inch from the bottom of the filter paper in PENCIL
Divide the filter paper into 4 parts by folding it into quarters
Each section (pigment) will have 3 sets of 3 drops
Using a Pasteur pipette, allow capillary action to draw up a drop of liquid and place on paper

7. Control Experimental
Or -carotene
Or Cholorphil B

8. Let the solvent run up the paper until it is an inch from the top
Immediately draw a line to indicate the solvent front and circle pigment bands
Do this in pencil

9. Measure distances in millimeters
If available, visualize the progression of the dyes using UV light source.
Write the RF values for the controls on the board
Use the averages of these in the lab report
Everyone needs to include all 4 controls even though you are only running 2 of them

10. Lab Report
Table with the averages of all the controls and the experimental Rf values for carrot and spinach
Also include S.D. and S.E.
Be sure to include all calculations for RF values
Use this information to determine which pigments are present in the spinach and the carrot