Activity 6 Paper Chromatography (Ink Analysis)

Analytical Chemistry (Qualitative and Quantitative Analysis) Laboratory Manual
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ACTIVITY 6
PAPER CHROMATOGRAPHY (INK ANALYSIS)
Chromatography is a simple technique used in separating substances in a
mixture and identifying those substances. The term derived from the words chroma
meaning color and graphein to write. There are three important components in
chromatography, namely:
1. the substances to be identified;
2. the moving phase, a solvent or solvents in which the solutes is
dissolved; and
3. the stationary phase, a filter paper, on which the compounds can be
absorbed.
In paper chromatography, component of ink is spotted on a piece of filter
paper. The filter paper is composed mostly of cellulose. Next an organic solvent is
drawn up the paper by capillary action. As the solvent moves over the location of the
ink, the ink begins to move up the paper. The rate at which the amino acid moves up
the paper is related to its relative affinity for the paper and the solvent.
I. Objectives:
To know some of the basic statistics that will be used in the laboratory;
To perform statistical analysis of the data obtained; and
To evaluate the probable magnitude of error in a measurement
II. Materials:
Chromatography paper
(either one sheet 10 cm x
20 cm, or single strips)
Beaker, 600 mL
Stapler
Watch glass/ plastic wrap
Activity 6 Paper Chromatography (Ink Analysis)
Isopropyl rubbing alcohol

Pencil
Ruler, 12 inch
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III. Procedure:
a.
b.
c.
d.
e.
f.
g.
Obtain a sheet of chromatography paper. Hold the paper by the edges

to avoid fingerprints which will interfere with the chromatography process.
There will be a pencil line on the chromatography paper about 1 cm from the
bottom. This line is called the origin. Also, on the line is a spot that was drawn
with a ball point pen. This spot is your unknown and was drawn with one of the
five pens you used.
In the laboratory, there is a set of five ball-point pens. Place a small
spot of ink from each pen on the pencil line. Space the spots so they are about
2 cm from the edge of the paper and about 2 cm apart. Directly above each
spot, label the paper, in pencil, with the name or identification of the pen used.
(See Figure I-1) The paper is already marked with the identification of the
unknown pen spot.
Roll the paper into a cylinder, butt the ends together, but do not overlap
them, and staple the paper together.
Add about 25 mL of isopropyl alcohol to the 600 mL beaker. Place the
chromatography paper into the beaker. Cover the beaker with a watch glass or
with plastic wrap. (See Figure I-2)
Allow the beaker and chromatography paper to sit for up to 30 minutes
for the inks to separate.
Do not disturb the beaker during this time. Do not handle the coins with
your fingers; always use the tweezers.
When the inks have separated sufficiently or the ink smears have
moved approximately 3/4 of the way up the paper, remove the
chromatography paper from the beaker. Open it flat and place it on a paper
towel to dry.
Unknown
Pen
Pencil line (origin)
Figure I-1. A sheet of chromatography paper

for analyzing ink from
several pens at one time.
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Figure I-2. The chromatography

paper placed in a container of
alcohol to separate inks.
Group No.__________ Date Performed: ___________________ Rating: ________

Year/Section: ____________________ Instructor: _________________________
ACTIVITY 6
PAPER CHROMATOGRAPHY (INK ANALYSIS)
IV. Data and Results:
Attach the chromatography paper to this sheet.
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Identification of the unknown spot: __________
V. Conclusion:
___________________________________________________________________
___________________________________________________________________
___________________________________________________________________
___________________________________________________________________
___________________________________________________________________
___________________________________________________________________
_________________________________________________________________.
Guide Question:
1. Does the ink from the unknown pen match any of the inks from the known
pens? Explain.

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Analytical Chemistry (Qualitative and Quantitative Analysis) Laboratory Manual