LABORATORY EXPERIMENT

pubs.acs.org/jchemeduc

Separation of the Components of a Commercial Analgesic Tablet: A

Two-Week Sequence Comparing Purification by Two-Base Extraction
and Column Chromatography
Kevin D. Revell*
Department of Chemistry, Murray State University, Murray, Kentucky 42071, United States
bS Supporting Information
ABSTRACT: A new laboratory experiment is described in which students
compare two benchtop separation methods to isolate the three active compo-
nents of the commercial analgesic Excedrin. In the two-week sequence, aspirin,
acetaminophen, and caffeine are separated using either a two-base liquid liquid
extraction or silica column chromatography. Students then evaluate the two
separation methods based on percent recoveries and on purity, as determined by
thin-layer chromatography. This procedure introduces three important techni-
ques using a single theme. The experiment has been designed to minimize cost of
chemicals and hazardous waste generation, while challenging students to
critically evaluate the strengths and weaknesses of different purification techni-
ques.
KEYWORDS: Second-Year Undergraduate, Laboratory Instruction, Organic
Chemistry, Hands-On Learning/Manipulatives, Acids/Bases, Chromatography,
Drugs/Pharmaceuticals, Separation Science, Thin Layer Chromatography

T he isolation of chemical compounds is a key component

of any undergraduate organic laboratory course. In addition
to recrystallization, distillation, and liquid liquid extraction,
chromatographic techniques such as thin-layer and column chro-
matography are of particular value to synthetic chemists. Although
several published experiments introduce preparative (column)
chromatography,1 3 the cost of materials and the volume of
waste produced make it difficult to teach this skill in an introductory
laboratory. A two-week laboratory sequence emphasizing chro-
matographic techniques has been developed and implemented
that involves the isolation of the three active components of the
commercial analgesic Excedrin. Students are able to evaluate the
utility of chromatographic techniques within the framework of
other purification methods.
In the first week, purification is accomplished by liquid liquid
extraction, using aqueous bases of different strengths to selectively
extract the acidic components. In the second week, students
purify the materials by column chromatography. The materials
isolated by each technique are analyzed and compared using thin-
layer chromatography (TLC). The students then evaluate and
compare the efficacy of the two techniques in terms of recoveries
and purity of materials obtained. These experiments have been
tested and modified over three semesters and have been conducted
by approximately 150 students.
Excedrin has been used in teaching labs to introduce a multitude
of techniques, including TLC,4 HPLC,5,6 capillary electrophoresis,7
solid-phase extraction,8 and GC and LC MS.9,10 However,
the use of two-base extraction11 and column chromatography for
the isolation of the three analgesic components appears to be
unique. This experiment is a valuable addition to the curriculum
because it introduces the students to three techniques and chal-
lenges them to critically evaluate the advantages and disadvantages
of each method. Furthermore, the materials are inexpensive and
only a small amount of hazardous waste is produced.
’ BACKGROUND
A single tablet of extra-strength Excedrin contains three active
compounds: aspirin (250 mg), acetaminophen (250 mg), and
caffeine (65 mg) (Figure 1). In addition, Excedrin contains a starchy
material that is used to bind the active components together. The
three analgesic components differ in terms of their acidity, with
aspirin being the most acidic (pKa 3.5), acetaminophen second
(pKa 9.9), and caffeine is effectively not acidic. The differences
in polarity of these three compounds produce good separations
on TLC, and a number of published procedures exist for the
separation of these analgesic components by TLC.1 4,12,13
’ EXPERIMENT
Week 1: Separation of the Excedrin Components by Two-
Base Extraction
The students crush one tablet of Excedrin (or the generic
equivalent) using a mortar and pestle, then add diethyl ether
(20 mL) to the powdered material (note: ethyl acetate may be
substituted for diethyl ether; see the technical notes in the
Published: July 27, 2011

Copyright r 2011 American Chemical Society and

Division of Chemical Education, Inc. 1413 dx.doi.org/10.1021/ed101195v | J. Chem. Educ. 2011, 88, 1413–1415
Journal of Chemical Education
Figure 1. The three active components in extra-strength Excedrin.
Supporting Information). The starchy material is insoluble in
organic solvents; however, a good portion of the active compo-
nents will dissolve in ether. The ether is decanted or filtered
through a conical filter into a separatory funnel. The solution is
then extracted successively with 1 M aqueous K2HPO4 (10 mL)
and 1 M aqueous KOH (10 mL). Because of the differences in
basicity, aspirin is extracted into the phosphate solution, and
acetaminophen is extracted into the hydroxide solution. Each of
these solutions is then acidified using 6 M aqueous HCl, causing
the conjugate acid to precipitate. Each component may then
be collected by filtration and air-dried. After extraction, the ether
layer contains primarily caffeine, which can be collected by evaporat-
ing the ether layer using a rotary evaporator or by traditional
vacuum distillation.
Following isolation, students determine the masses and cal-
culate the percent recoveries for each component. Typically, the
recoveries of aspirin and acetaminophen improve if the acidic
solutions are allowed to stand; thus, students store these solutions in
their lab drawers until the next period, when they complete the
filtration and conduct the TLC analysis described below.
Week 2: Separation of the Excedrin Components by
Chromatography
Part A: Thin-Layer Chromatography. In the first part of
week 2, students compare the purified components of Excedrin
(stock solutions of the purified components are provided by the
instructor) with the mixture obtained when a tablet is crushed,
dissolved in ether, and filtered. The students obtain good results
by TLC when they use small (2  5 cm) glass-backed plates with
a UV indicator, use a 30 mL beaker with a watch-glass lid as a
developing chamber, use commercial micropipets, and make very
small spots on the plates. The students develop the plates using a
solution of 1:2 hexanes/ethyl acetate with 1% acetic acid as
eluent. Three distinct spots having Rf values of 0.6 (aspirin), 0.3
(acetaminophen), and 0.1 (caffeine) are observed for the mixture
under these conditions (visualized under UV light).
Part B: Column Chromatography. In the second part of
the week 2 experiment, students prepare an ether solution
containing the active components of the two tablets, using a
procedure identical to part A. This solution is placed in a small
beaker, and a small volume (0.5 mL) of silica gel is added. The
solution is stirred as the ether is evaporated under a gentle stream
of air and slight warming, resulting in a white solid containing the
three active components adsorbed onto silica. This solid is loaded
onto a small polypropylene column containing 3 g of silica. The
silica is then flushed with solvent to elute the three products.
Although all three products can be eluted with a single solvent
mixture, successive addition of 1:1 hexanes/ethyl acetate (30 mL),
1:2 hexanes/ethyl acetate (30 mL), and acetone (30 mL) elutes
all three components quickly and with good separation. When
15 mL capacity test tubes are used, the three components can
be collected and resolved using only 6 tubes. The contents of
each tube are determined using TLC. The students used rotary
1414
LABORATORY EXPERIMENT
Figure 2. TLC of Excedrin components after separation, showing aspirin
(Rf = 0.6), acetaminophen (Rf = 0.3) and caffeine (Rf = 0.1). Eluted with
33:66:1 hexanes:ethyl acetate:acetic acid. (A) Results from week 1: (1)
acidified phosphate extracts, (2) acidified hydroxide extracts, and (3)
organic layer. (B) Results from week 2: column eluent, tubes 1 6.
evaporators to concentrate their solutions and isolate the materials.
The students then measure the masses and calculate the percent
recoveries for each component.
Part C: Comparison of the Two Preparative Techni-
ques. In the last step, students use TLC to analyze the three
components that were isolated from the two-base extraction
(Figure 2A). They compare the purity of the materials obtained
by extraction and by column chromatography (Figure 2B). Addi-
tionally, the students compare the yields obtained in each step.
’ HAZARDS
Goggles should be worn at all times in the laboratory. This
experiment involves strong acids and bases, which are corrosive.
Diethyl ether, ethyl acetate, and hexanes are each flammable and
should be handled in fume hoods. The students should especially
exercise care when working with diethyl ether, which is highly
volatile. If time and resources permit, the instructor may use ethyl
acetate or dichloromethane in place of ether (see technical notes
in Supporting Information). The MSDS should be consulted for
all materials prior to conducting this experiment.
’ TYPICAL RESULTS
Generally, students are able to obtain aspirin in very high
recoveries (60 100%) in both procedures. Recoveries for
acetaminophen are often substantially lower (0 10% for extrac-
tion, 10 20% for chromatography) because the acetaminophen
often forms a fine solid that is difficult to filter. Caffeine typically
is recovered in higher percentages in the chromatography (40%)
than in the extraction (1 5%) (see technical notes in Supporting
Information).
’ SUMMARY
The experiment described involves the use of three important
techniques (extraction, column chromatography, and TLC) to
isolate and analyze the components of Excedrin. Although some
capital equipment must be available to conduct the experiment,
dx.doi.org/10.1021/ed101195v |J. Chem. Educ. 2011, 88, 1413–1415
Journal of Chemical Education LABORATORY EXPERIMENT

the chemical components of the experiment are fairly inexpensive,

and the amount of waste generated is modest.

’ ASSOCIATED CONTENT

bS Supporting Information
A list of required equipment and chemicals; a detailed experi-
mental procedure; and notes for the instructor. This material is
available via the Internet at http://pubs.acs.org.

’ AUTHOR INFORMATION
Corresponding Author
*E-mail: Kevin.revell@murraystate.edu.

’ ACKNOWLEDGMENT
The author gratefully acknowledges Beth Brubaker for her
excellent management of the teaching laboratories at Murray
State University and for her assistance in acquiring the elements
needed to develop these experiments. The author also is grateful
to Carl Woods and Daniel Johnson, who assisted with the TLC
images above. Carl Woods also produced the artwork included in
the student handout.

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