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Chromatography Lab Worksheet

The document outlines a chromatography lab worksheet focused on separating and identifying components of mixtures using paper chromatography. It explains the principles of chromatography, including the roles of the stationary and mobile phases, and introduces the concept of retention factors (Rf) to quantify separation. The lab aims to determine whether the colors in marker dyes are due to single compounds or mixtures, with specific objectives, materials, and procedural steps provided.

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Marco Antonio Paglinawan Paredes
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5 views

Chromatography Lab Worksheet

The document outlines a chromatography lab worksheet focused on separating and identifying components of mixtures using paper chromatography. It explains the principles of chromatography, including the roles of the stationary and mobile phases, and introduces the concept of retention factors (Rf) to quantify separation. The lab aims to determine whether the colors in marker dyes are due to single compounds or mixtures, with specific objectives, materials, and procedural steps provided.

Uploaded by

Marco Antonio Paglinawan Paredes
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd
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Chromatography Lab Worksheet

Background: Almost all substances we come into contact with on a daily basis are impure; that is, they are
mixtures. Similarly, compounds synthesized in the chemical laboratory are rarely produced pure. As a result, a
major focus of research in chemistry is designing methods of separating and identifying components of
mixtures. The theory behind chromatography is to allow a mixture of different chemicals to be distributed or
partitioned between a stationary phase and a mobile phase (eluent or solvent). The mobile phase may be a
liquid or a gas; the stationary phase is typically a solid.

As the mobile phase flows over the stationary phase, the components in the mixture are carried along. The
more soluble a component is in the mobile phase the faster it will be transported along the stationary phase.
Adsorption refers to the ability of a substance to ‘stick’ (or be adsorbed) to a surface. The more strongly a
component is adsorbed to the stationary phase, the slower it will be transported by the mobile phase.

As the mixture moves over the stationary phase, the components in the mixture move further and further
apart into discrete zones. The purpose of this lab is to utilize paper chromatography to determine whether the
colors of marker dyes coloring are due to a single compound or a mixture of several other colors.

Retention Factors (Rf): The affinity of a substance for the stationary and mobile phases is characteristic of that
substance. Different substances will have different competitive affinities. Since each component of a mixture
will have its own characteristic affinities, each component will travel up the paper at its own characteristic
rate. If the paper is sufficiently large, all the components can be separated by the time the solvent front has
reached the top of the paper and each component will appear as a separate spot.

The chromatographic paper will now contain a vertical array of colored spots arranged according to their
characteristic rates of ascent. It is possible to describe the position of spots (so the substances that have
separated) in terms of their retention factor, the Rf value (Figure 1). The retention factor is defined as: Rf =
distance traveled by spot / distance traveled by solvent.

Objectives:
 Describe how chromatography works and what happens during this process.
 Explain why being able to separate solutions into their components is important.

Materials:
 Whatman chromatography paper, at least 4 two-inch-wide strips; available at amazon.com
 pencil
 Sharpie® black ink marker; can be shared among teams
 Sharpie color markers, of various colors; can be shared among teams
 isopropyl alcohol (rubbing alcohol); higher proof or percent rubbing alcohol is best
 two 500-ml beakers

Procedure:
1. Take the strips of chromatography paper and fold both pieces about an inch from the top.
2. Using the black Sharpie, draw a horizontal line near the other end of the chromatography paper. Make the
line about as high off the bottom as the length of your thumb.
3. Use a pencil to suspend the paper in each beaker. Make sure the strip does not touch the sides of the
beaker.
4. Carefully add water to one beaker and alcohol to the other. Add just enough of each liquid so that it
touches the bottom of the hanging strip.
5. Watch what happens!! Once you see the ink separation is complete, get new strips and repeat using ink
from a colored Sharpie marker.

Analysis Questions:

1. What were the two solvents used?

2. Black ink is more attracted to which solvent? How do you know?


3. What colors are present in black ink?

4. What are the two major phases of chromatography? Describe each phase.

5. Explain in your own words why samples can be separated into their components by chromatography.

6. Calculate the retention factors (Rf) for both of your samples using the methods described below and in
figure 1. You will need to show all of your work and use the correct significant figures.

7. If two components have the same Rf value in a chromatogram, are they necessarily identical in structure?
Please justify your response

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