Name: _____________________________ Rating: ___________

STEM 12- _____ Group No. _____ Date: _____________

Activity 5
Let’s Separate

Objectives
 Describe separation techniques for the mixtures and compounds
 Apply simple separation techniques such as distillation and chromatography.

Pre- Lab Discussion

Chromatography is a laboratory technique for the separation of a mixture. The mixture is dissolved in a fluid called
the mobile phase, which carries it through a structure holding another material called the stationary phase. The various
constituents of the mixture travel at different speeds, causing them to separate. The separation is based on differential
partitioning between the mobile and stationary phases. Subtle differences in a compound's partition coefficient result in
differential retention on the stationary phase and thus affect the separation.

Materials
Ethanol, Acetone, Water, Filter paper/ Chalk, beaker

Procedure

1. Apply your ink to a piece of chalk about 1cm from the end of the chalk. You can place a dot of color or a band of color
all the way around the chalk.
2. Pour enough isopropyl alcohol/ acetone into the bottom of the jar or cup so that the liquid level is about half a cm
high. Make sure the liquid level is below the dot or line of color on your chalk.
3. Place the chalk in the beaker. Leave the chalk for about several minutes and watch the color climb up to the chalk.
4. Look closely at each marker’s chromatography strip. How many different colors are present in each ink sample?
Record the Total Number of Colored Pigments number for each ink sample on your Reference Library.
5. Look closely at each marker’s chromatography strip. Record the colors in the order that they appear on your
Reference Library page. Each color represents a different pigment present in the ink. (Colored Pigment #1 = pink,
Colored Pigment #2 = orange, etc.)
6. To prove that an ink sample is a certain brand, you will also need to calculate the Rf (retention factor) values of the
different colored chemicals present in the marker. Rf is a calculation that compares the distance the solvent traveled
up the paper strip to the distance a pigment traveled up the same strip. First, look at a chromatography strip and
measure the distance in millimeters from the original color dot to the final point the solvent traveled. That distance is
the solvent distance measurement. Record it on your Reference Library page.
7. Next, measure in millimeters from the original color dot to the highest point the first colored pigment (Colored Pigment
#1) traveled up the strip. This is the pigment distance measurement for Colored Pigment #1. Record this
measurement on your Reference Library page.
8. If there is a Colored Pigment #2 on the strip, measure in millimeters the distance from the original color dot to the
highest point this colored pigment (Colored Pigment #2) traveled up the strip. This is the pigment distance
measurement for Colored Pigment #2. Record this on your Reference Library page.
9. Repeat the measurement for any other colored pigments and record on your Reference Library page.
10. Calculate the Rf value for each colored pigment using the formula: Rf Value = Distance traveled by solute (pigments
in ink) / Distance traveled by solvent (alcohol).
11. Record the Rf values for each colored pigment on your Reference Library page.
12. Repeat these measurements and Rf calculations for each of the markers tested.
13. Answer the follow-up questions.
Data and Results

REFERENCE LIBRARY FOR INK SAMPLES AND THEIR COLORED PIGMENTS

Color of the Pigment ______ Color of the Pigment ______ Color of the Pigment ______
Total # of Colored Pigments _____ Total # of Colored Pigments _____ Total # of Colored Pigments _____
Solvent Distance Measured _____ Solvent Distance Measured _____ Solvent Distance Measured _____
Colored Pigment #4 Colored Pigment #4 Colored Pigment #4
Color __________ Color __________ Color __________
Distance measured __________ Distance measured __________ Distance measured __________
Rf _________ Rf _________ Rf _________
Colored Pigment #3 Colored Pigment #3 Colored Pigment #3
Color __________ Color __________ Color __________
Distance measured __________ Distance measured __________ Distance measured __________
Rf _________ Rf _________ Rf _________
Colored Pigment #2 Colored Pigment #2 Colored Pigment #2
Color __________ Color __________ Color __________
Distance measured __________ Distance measured __________ Distance measured __________
Rf _________ Rf _________ Rf _________
Colored Pigment #1 Colored Pigment #1 Colored Pigment #1
Color __________ Color __________ Color __________
Distance measured __________ Distance measured __________ Distance measured __________
Rf _________ Rf _________ Rf _________

Questions
1. How many colors separated from the black ink? List the colors from top to bottom.

2. What do you observe from the different colors of pen?

3. What served as the solvent for the ink?

4. Why the inks separate?

5. Why did some inks move a greater distance?

Conclusion