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Lesson 1: Chromatography
Chromatography is used to determined what compounds
make up an unknown sample. Samples are mixed with a gas or liquid,
then passed across another substance which your sample is differentially
attracted to. The compounds are sorted and separated, those with
weaker attractions traveling faster and farther than those with
strong attractions. These results are compared to tests of known
substances to determine what the unknown sample is made of.
Chromatography can be used for many tasks, including: separating
plant pigments, detecting accellerants in arson investigations,
determining if there is DDT in bird eggs, analyzing ink samples
in detective investigations, determining the paint composition and
possibly the authenticity of a painting. In this lesson, we will
be using liquid chromatography to test ink samples but this will
help students understand the process and its relationship to testing
eggs for DDT.
Time Allotment:
one 45-minute session
Materials:
- strips of paper towel or coffee filter (cut all to same width)
- 6 sets of 4-5 different ink (felt tip) pens (same color, different
brands)
- 4-5 plastic cups or a single shallow tray per group
- water
- salt
- 4-5 coffee stirrers or sticks
- 6 copies of writing "sample" ink (on test strip)
- 6 copies of writing "sample" (can be a made-up poem or a copy
of a portion of a work actually from the time in question)
Advanced Preparation and Teacher Notes
- Prepare filter strips with "sample" of mystery ink
- Test felt tip pens selected to be sure they will result in
a different pattern in the chromatograph
- Arrange students in cooperative groups of four
Tap Prior Knowledge
Explain to the students that you have a mystery to solve. They
are archivists working at a museum. Someone has come in with a manuscript
that he claims is from a famous author who lived in the 1600's.
It is your job to determine if the manuscript is really that old.
Ask the students how they would do this. Write their answers on
the board. Explain that for now, cost (or equipment) is no object.
Discuss the pros and cons of each suggestion as well as what it
might tell them about the manuscript in question. Be sure to mention
when a technique might damage or destroy the object being tested
(and why this is not such a good idea).
Share with Neighbor
Have the students group into pairs to study the sample and make
a hypothesis. Is it authentic? Why do they think so? If they are
having trouble, you can encourage them by explaining that even without
scientific equipment, they might be able to make an educated guess
using things like how old the paper looks (lined paper was not around
in its current form in the 1600's) or what words are used and how
they sound.
Engage Students in Hands-on Activity
Now even though we don't have a lot of equipment to do many of
the scientific tests actual museum curators would use, we do have
access to one method. We can find out what ink might have been used.
Pass out the "sample inks" (i.e., Pens labeled 1-5). Each pen contains
ink from a specific time period. List these on the board. Any dates
will do so long as the one that will match the writing sample has
the correct date: the 17th century.
Have one person in the group collect the remaining supplies needed
for the experiment: a shallow tray or 5 plastic cups, the strips
of coffee filter, and the "sample" strip with the line of unknown
ink in it.
Have each group draw a line one inch from the bottom with each
of the inks to be tested on the strips of coffee filter. One ink
sample (pen) per strip.
Each strip should then be taped to a coffee stirrer at the non-marked
end.
Place each strip into the water so that the edge is submerged,
but do not submerge to the ink line. Students using cups can place
the coffee stirrers across the rim of the cup to hold the strips
vertical. When using a tray, pile textbooks on either side to hold
the stirrers.
Have the students record what happens in their journals
When the inks have finished separating have them match the sample
inks (pens) to the sample from the unknown writing.
Which sample 1-5 matched our written work?
Compare dates of each sample to determine when the work was written.
Introduce the Scientific Principle/Concept
When scientists want to find out what a substance is made of, one
of the techniques they can use is chromatography. Chromatography
separates substances into their component parts. For example, it
would separate the chlorophyll (the green pigment in plants) of
a leaf into chlorophyll a, chlorophyll b, and other pigments. Chromatography
works because the atoms and molecules of a given compound are attracted
with different strengths to other atoms and molecules. Chromatography
pushes the substance tested across another stationary substance
for which it has an attraction. Molecules with a stronger attraction
to the stationary substance are slowed down more than those with
a weaker attraction. A measuring device is used to determine what
substances are present based on how far they have traveled in a
given time, or how fast they arrive at the detector after being
placed in the chromatograph.
Typically, the substance to be tested is pushed past the stationary
substance in a liquid or a gas, which acts as a "carrier." In our
experiment, we used liquid chromatography, where our line of ink
was separated by a liquid and the component parts carried up a piece
of paper. How far up the paper each dye (or component) in the ink
traveled depended on how attracted each dye was to the wet paperthose
with stronger attractions did not travel as far. This left bands
of color, one band for each dye. Each pen, though producing black
ink, is really made up of a number of dyes in combination. Not every
ink uses the exact same combination of inks. That's why when they
were allowed to separate only one matches our samplethe ink
that was used to write it.
Chromatography can be used in other scientific fields to identify
chemicals. Gas chromatography was used to detect DDT amounts in
bird eggs.
Relate Activity and Concept
Have the students try chromatography on different color ink pens.
Were the colors used to make each pen what they expected? Which
color had ink components that traveled he farthest? The shortest?
Did the same pigments (component colors) always travel the same
relative distance?
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