In the Laboratory

Salicylate Detection by Complexation with Iron(III)

and Optical Absorbance Spectroscopy
An Undergraduate Quantitative Analysis Experiment
Jeremy T. Mitchell-Koch*
Department of Chemistry, Emporia State University, Emporia, KS 66801; *jmitchel@emporia.edu

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Publication Date (Web): December 1, 2008 | doi: 10.1021/ed085p1658

Kendra R. Reid and Mark E. Meyerhoff

Department of Chemistry, University of Michigan, Ann Arbor, MI 48109

The analysis of common over-the-counter medications

has become a popular approach in undergraduate analytical
chemistry laboratories (14). The connection of the students
course work to the real world is exciting and often leads to
an increased interest in chemical analysis. This idea has been at
the heart of the quantitative analysis laboratory courses at both
Emporia State University and the University of Michigan, and
our experiences led us to believe that our approach improves
student learning and student enthusiasm for carrying out given
laboratory experiments. Spectroscopy is a critical tool in the
analysis of different molecules and is incorporated in the secondyear analytical chemistry laboratories. This experiment describes
the quantitation of salicylate in liquid face wash using visible
spectrophotometry. A similar colorimetric method published
in this Journal described the detection of salicylate in urine
for nursing students (5). Here, we expand the application of
visible spectroscopy to determine salicylate in commercial face
wash products and also to investigate the nature of the colored
product formed during the analysis.
There are several uses for salicylate and it is included in
many everyday products. Salicylic acid is the major metabolite
of aspirin and is commonly found in medications that treat
acne, warts, and other similar ailments. Owing to the many
medical applications of salicylic acid, a number of methods
for its quantitation have been developed, including titration
(6), gasliquid chromatography (7), ultraviolet spectroscopy
(8), and fluorescence spectroscopy (9, 10). The most widely
used method in clinical laboratories, however, employs visible
spectrophotometry where excess iron(III) is added in an acidic
solution to form a highly colored species (11).
A version of the visible spectrophotometry method is applied to quantitate salicylate in a commercial product and also
in an instructor-prepared unknown solution. The BeerLambert
law is followed over the concentration range of interest (10100
mM), and a linear calibration curve is prepared. For the analysis
of the commercial face wash, we collected and posted all trials
for the entire class and asked students to conduct a t test comparing their data to that of the class. In addition, students compared their results with the salicylic acid value provided by the
manufacturer. This requires students to practice unit conversions
from amount concentration to mass concentration expressed as
a percentage. Typical student results for this analysis compare
favorably to the salicylic acid level that the manufacturer claims.
A recent class at Emporia State University (8 students, 24 analyses) determined an average salicylic acid concentration of (1.01
0.09)% compared to 1% listed by the manufacturer (only one
significant figure provided). While these results generally confirm the accuracy of the analysis, synthetic unknown solutions
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are also used to further assess the students analytical skills. In

these experiments all students generally determine the unknown
concentration to within 5% of the expected value. These results
indicate that students benefit from the opportunity the experiment provides to learn about a critical analytical tool.
In the second part of the experiment, the method of
continuous variation ( Jobs method) is used to investigate
the stoichiometry of the ironsalicylate coordination complex formed. Varying volumes of 10 mM solutions of sodium
salicylate and iron(III) nitrate are mixed and diluted to prepare
eleven test solutions (see Table 1 in the online material). The
total combined volume of the two solutions remains constant
(1 mL), and thus only the mole ratio of iron to salicylate changes
in these solutions. The test solution that exhibits the largest
absorbance at max indicates the mole ratio of the predominant
complex formed in the reaction. This maximum is most easily
demonstrated in a plot of absorbance versus mole fraction of
iron(III).
Experimental Procedure
Spectrophotometric Determination of Salicylate
in Acne Medication
A stock solution of approximately 100 mM salicylate is
prepared from its dried sodium salt and is then appropriately
diluted to yield five other standards for the calibration curve. To
100 L of each standard, triplicate samples of face wash, and the
synthetic unknown, 10.00 mL of acidic iron(III) nitrate solution
is added. The absorbance of these solutions is then measured
in plastic cuvettes. This step may be modified depending on
available instrumentation. In one class, each pair of students
had access to an Ocean Optics absorbance instrument that
provided the entire spectrum for each sample. This allows for
simple determination of max and the absorbance values at max.
In another laboratory setting, one scanning spectrophotometer,
and several single wavelength spec 20 units were available. In
this situation, each pair of students collected a spectrum of one
standard to determine max. Then the spec 20 units were set to
this wavelength and the absorbance of each calibrant and test
solution was determined. If only single wavelength instruments
are available, instructors may provide students with the max
value to expedite the measurement process (~535 nm). From the
absorbance data collected for the standards, a linear calibration
curve is constructed (Figure 1), and the concentration of each
unknown is determined using a least-squares fit line derived from
the calibration data. The absorbance data are plotted against the
undiluted standard concentrations to simplify quantitation of
salicylate in each unknown. Since all standards and unknowns

Journal of Chemical Education Vol. 85 No. 12 December 2008 www.JCE.DivCHED.org Division of Chemical Education

In the Laboratory

Determination of Reaction Stoichiometry:

An Application of the Method of Continuous Variation
Various volumes of 10 mM iron(III) nitrate and 10 mM
sodium salicylate are mixed in separate test tubes in ratios shown
in Table 1 (found in the online material). The total volume of
these two solutions is a constant 1.00 mL, keeping the total
amount of reagents the same. Dilute nitric acid (4.00 mL, ~60
mM) is added to each mixture, and the absorbance at max is
recorded for each solution. A typical plot of absorbance versus
the mole fraction of iron(III) is shown in Figure 2. An apparent
local maximum around 0.250 mole fraction (a 1:3 iron:salicylate
complex) is a typical result, which may represent a minor complex, but the overall maximum in this plot at 0.500 mole fraction (a 1:1 iron:salicylate complex) indicates the predominant
stoichiometry of the reaction.

Absorbance at 535 nm

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Hazards
Nitric acid is corrosive, and iron(III) nitrate is an oxidizer:
appropriate safety precautions should be taken when handling
these chemicals.
Summary
The experiment provides students practical experience
with BeerLambert law for the spectroscopic quantitation of
molecules in aqueous solution. In addition, spectroscopy is used
to investigate the nature of a chemical reaction and determine
the stoichiometry of the absorbing species. The experiment
can be completed in a single three-hour laboratory session but
may be divided to provide students ample time to analyze their
data as it is collected. The reagents are common, inexpensive,
and require no special safety precautions. The experiment can
be easily adapted to a wide variety of spectrophotometers and
therefore is simple to adopt into any second-year quantitative
analysis course.
This experiment is relatively simple to perform and helps
students gain an understanding of optical spectrophotometry
and equilibrium. If appropriate analytical techniques are used,
students encounter little difficulty in obtaining a linear calibration curve and determining the predominant complex stoichiometry. Student results demonstrate excellent accuracy (vide
supra), and this enhances the enjoyment of the activity for both
students and instructors.
Acknowledgments

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The authors acknowledge the University of Michigan for

funding and all the students at U of M and ESU who tested
this experiment.

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Literature Cited

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20

40

60

80

Salicylate Concentration / (mmol/L)

Figure 1. Typical calibration curve for salicylate.

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3.
4.
5.

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7.
8.
9.
10.
11.

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Absorbance at 535 nm

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Publication Date (Web): December 1, 2008 | doi: 10.1021/ed085p1658

are diluted in the same manner, the salicylate concentrations in

the unknown solutions can be extracted directly from the calibration curve without any dilution calculations. This method is
valid for concentration determination, but the plot will underestimate the extinction coefficient for the ironsalicylate complex
by approximately two orders of magnitude. The experiment
may be modified to require a plot of absorbance versus actual
salicylate concentration (after dilution in the reaction mixture)
to properly determine the extinction coefficient.

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Ferguson, G. K. J. Chem. Educ. 1998, 75, 467469.

Hein, J.; Jeannot, M. J. Chem. Educ. 2001, 78, 224225.
Simonson, L. A. J. Chem. Educ. 2001, 78, 1387.
Yang, S.-P.; Tsai, R.-Y. J. Chem. Educ. 2006, 83, 906909.
Cavanaugh, M. A.; Bambenek, M. A. J. Chem. Educ. 1978, 55,
464.
Lane, S. R.; Stewart, J. T. J. Chem. Educ. 1974, 51, 588589.
Battezzati, A.; Fiorillo, G.; Spadafranca, A.; Bertoli, S.; Testolin,
G. Anal. Biochem. 2006, 354, 274278.
Rogic, D. J. Mol. Struc. 1993, 294, 255258.
Lange, W. E.; Bell, S. A. J. Pharm. Sci. 1966, 55, 386389.
Saltzman, A. J. Biol. Chem. 1948, 174, 399404.
Annino, J. S.; Giese, R. W. Clinical Chemistry: Principles and
Procedures, 4th ed.; Little, Brown and Co.: Boston, 1976; pp
355357.

Supporting JCE Online Material

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http://www.jce.divched.org/Journal/Issues/2008/Dec/1658.html
Abstract and keywords

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Full text (PDF) with links to cited JCE articles

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Mole Fraction Fe(III)

Figure 2. Typical Job Plot for the iron-salicylate complex.

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Supplement

Student handouts

Instructor notes

Division of Chemical Education www.JCE.DivCHED.org Vol. 85 No. 12 December 2008 Journal of Chemical Education

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