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In the Laboratory

Isolation of Shikimic Acid from Star Aniseed


Richard Payne
Department of Chemistry, University of Canterbury, Private Bag 4800, Christchurch, New Zealand
Michael Edmonds*
School of Applied Science, Christchurch Polytechnic Institute of Technology, PO Box 540, Christchurch 8015,
New Zealand; *edmondsm@cpit.ac.nz

The isolation of natural products from plant material is


a common way of introducing students to some of the key
concepts in organic chemistry. Consequently many undergraduate chemistry courses include experiments such as the
isolation of eugenol from cloves (1) or caffeine from tea (2).
However in developing a course specifically aimed at teaching natural products chemistry a greater range of experiments
is required. While several useful resources are available (3,
4), we were interested in developing additional experimental
procedures in the area of natural product chemistry. Consequently this article describes a new undergraduate laboratory
experiment suitable for demonstrating some key techniques
used in natural products chemistry.
Background
Shikimic acid (Figure 1) plays a key role in the biosynthesis of many important natural products including aromatic
amino acids, alkaloids, phenolics, and phenylpropanoids. Indeed, its role is of such importance that one of the key biosynthetic pathways is referred to as the shikimate pathway.
Shikimic acid was first isolated in 1885, from parts of
the Japanese tree, shikimino-ki (Illicium anisatum). However
it is also present in the seeds of the fruit of the star aniseed
(Illicium verum). This experiment describes the isolation of
shikimic acid from star aniseed in up to 7% yield and is
adapted from the work of Adams et al. (5). The article by
Adams and co-workers describes the extraction of 900 g of
aniseed using a 24 h Soxhlet extraction and subsequent purification using anion exchange and Solka-Floc. In adapting
this experiment for teaching purposes we have shortened the
extraction time and attempted to introduce more common
purification techniques, for example, the use of decolorizing
charcoal. The isolation of shikimic acid from Eucalyptus
citriodora is also possible (6).
The presence of the carboxylic acid functionality in the
structure of shikimic acid allows ion exchange chromatography to be utilized in the purification of shikimic acid. Other
techniques used in this practical experiment include Soxhlet
extraction and recrystallization techniques.
Experimental

COOH

HO

OH
OH

Figure 1. Structure of shikimic acid.

brown colored filtrate under reduced pressure using a rotary


evaporator afforded a thick brown oil (approximately 8.5 g),
which was redissolved in water (145 mL) and warmed to 80
C. Any dark green oil that formed on the surface of the aqueous solution was removed by Pasteur pipet. Approximately
five drops of 3740% formaldehyde solution was added to
the hot solution and the aqueous solution was boiled for 5
min then allowed to cool. The resulting precipitate was removed by passing the solution through a glass filter funnel
containing a layer of celite to afford a clear orange solution.
The solution was passed through an anion exchange column
(Amberlite IRA-400, in acetate form, dry weight 25 g). The
column was then washed with water (100 mL), which was
discarded. The column was then eluted with acetic acid (185
mL of 25% acetic acid in water) and the yellow eluent was
collected. Removal of the acetic acid by rotary evaporation
followed by high vacuum pump afforded an orange colored
solid (approximately 3.0 g).
The solid was dissolved in methanol and heated for 10
min with activated charcoal (3 spatulas). Filtration and subsequent removal of the methanol under reduced pressure afforded an off-white solid (approximately 2.0 g). The solid
was then recrystallized from methanol and toluene (or ethyl
acetate) to afford the desired shikimic acid as a bright white
crystalline solid. Typical yield were between 0.60 and 1.74 g
(2.4 to 7.0%). Analysis by melting point and NMR (5) confirmed the purity of the shikimic acid.
Analyses
Melting point: 184185 C (lit. 184 C).

Approximately 25 g of star aniseed was ground using a


blender and accurately weighed into a Soxhlet thimble. The
thimble was then placed in the Soxhlet apparatus and heated
under reflux with 95% ethanol (125 mL) for approximately
2 h (by which time the color of the solvent running from
the thimble was colorless). Removal of the ethanol from the

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1H

NMR (300 MHz, D2O, ): 6.70 (m, 1H), 4.30 (m,


1H), 3.93 (m, 1H), 3.67 (dd, J = 8.4, 4.5 Hz, 1H), 2.64
(dd, J = 18.0, 4.8 Hz, 1H), 2.12 (dd, J = 18.0, 6.3 Hz,
1H).

13C

NMR (75 MHz, D2O, ): 170.1, 137.1, 129.8,


75.1, 66.5, 65.8, 30.4.

Vol. 82 No. 4 April 2005

Journal of Chemical Education

599

In the Laboratory

Hazards

(one or two sessions of 34 hours) and is appropriate for both


organic and natural product courses.

No hazardous data are reported for Amberlite IRA-400


resin. Ethanol is highly flammable. Formaldehyde solution
is toxic and can cause burns. Acetic acid has a pungent smell,
can cause burns, and is irritating to respiratory system. Concentrated hydrochloric acid causes burns and is irritating to
respiratory system. Sodium acetate is classified as nonhazardous. Activated charcoal is harmful by inhalation and irritating to eyes, respiratory system, and skin. Celite is harmful
and is irritating to eyes and respiratory system. There is danger of serious damage to health by prolonged exposure
through inhalation. Methanol is highly flammable and toxic
by inhalation, in contact with skin, and if swallowed. Toluene is highly flammable and harmful by inhalation. Ethyl
acetate is highly flammable and harmful by inhalation.
Conclusion
A laboratory experiment has been developed which in
the process of extracting shikimic acid from star aniseed exposes students to a number of important experimental techniques. The experimental procedure is designed to be carried
out in a time period suitable for most undergraduate courses

Supplemental Material

Information for the instructor, which includes an introduction to the experiment, a list of experimental conditions
and requirements, 1H and 13C spectra, and a handout for
students are available in this issue of JCE Online.
Literature Cited
1. Ntamila, M. S.; Hassanali, A. J. Chem. Educ. 1976, 53,
263.
2. Hampp, Andreas. J. Chem. Educ. 1996, 73, 1172.
3. Ikan, R. Natural ProductsA Practical Guide; Academic Press:
New York, 1991.
4. Dickson, C. Medicinal Chemistry Laboratory Manual; CRC
Press: Boca Raton, FL 1999.
5. Adams, H.; Bailey, N.; Brettle, R.; Cross, R.; Frederickson,
M.; Haslam, E.; MacBeath, F.; Davies, G. Tetrahedron 1996,
52, 85658580, and references therewithin.
6. Anet, E. F. L.; Birch, A. J.; Massy-Westropp, R. A. Aust. J.
Chem. 1957, Vol. 10, 9394.

The structure of shikimic acid discussed in this article is available in fully manipulable Chime format
as a JCE Featured Molecule in JCE Online (see page 656).

JCE Featured Molecules

an interactive modeling feature, Only@JCE Online

http://www.JCE.DivCHED.org/JCEWWW/Features/MonthlyMolecules

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Journal of Chemical Education

Vol. 82 No. 4 April 2005

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