Name: ____________________________

Chromatography Lab Worksheet

Pre-Lab Questions

1. Figure 1 is a sample paper chromatogram for three

samples: A, B and C. Sample C gave two spots on
the paper chromatogram. What does this tell you
about the composition of the sample?

2. Based on the Rf values of samples A and B, what can you conclude about the intermolecular
attractions both samples have for the solvent and the paper?

3. Examine the structures of the FD&C Red No. 40, Blue No. 1 and Yellow No. 5 dyes (on the
procedure page). What are the similarities and differences in the structures of the three dyes?
Talk specifically about number of charged sites (not including Na+) and number of sites
capable of participating in hydrogen bonding.

Dye Number of charged sites Number of sites that can hydrogen

bond

Red No. 40

Blue No. 1

Yellow No. 5

4. Briefly describe column chromatography and how it works. Use the terms stationary phase,
mobile phase, and affinity in your description.
 Name: ____________________________

5. Two of the solvents we will use in this lab are 0.1% aqueous sodium chloride solution and 2%
aqueous isopropyl alcohol solution. Draw separate molecular diagrams of how sodium
chloride and isopropyl alcohol would interact with water. Identify the types of intermolecular
attractions within each diagram.

Part 1 – Observations

Solvent Used Observation of Fraction Collected

Water

2% isopropanol

70% isopropanol

6. Based on the results from part 1, which of the three dyes was the most polar? Which dye
was the least polar? Explain how you know, making reference to your observations as
well as the polarities of both the column and the solvent.
 Name: ____________________________

Part 2 – Data

Solvent Used Distance traveled Distance traveled Distance traveled Distance traveled
by red by blue by yellow by solvent
Water

0.1% NaCl

20% NaCl

Part 2 – Calculation and Questions

7. Fill in the chart below with Rf values for each spot in each solvent (show work)
Solvent Used Red dye Rf Blue dye Rf Yellow dye Rf
Water

0.1% NaCl

20% NaCl

8. Which solvent in part 2 gave the best separation of the dyes? How did you judge this?

9. Based on the results from part 2, which of the three dyes has the most attraction for
cellulose (the material the paper is made from)? Explain how you know, making
reference to your observations as well as the types of interactions that the dyes would
have with cellulose. FYI – cellulose is a long polymer chain of glucose molecules.
Glucose molecules are cyclic with many OH groups around the ring.
 Name: ____________________________

Part 3 – Observations
Draw your chromatogram(s) using color and label your drawing. Unknown ____________

Part 3 – Questions

10. What dyes were mixed together to make your unknown mixture? Explain.

11. Propose an explanation for why 0.1% NaCl separated the dyes better than water or 20%
NaCl did. Your answer will not be graded for correctness but instead on whether it is
clear that you thought about this. Some things to think about include how number of
charged sites and number of hydrogen bonding sites influenced a dye’s ability to travel in
NaCl(aq).