Laboratory Experiment

pubs.acs.org/jchemeduc

Synthesis of a D-Glucopyranosyl Azide: Spectroscopic Evidence for

Stereochemical Inversion in the SN2 Reaction
Olumuyiwa G. Adesoye, Isaac N. Mills, David P. Temelkoff, John A. Jackson,* and Peter Norris*
Department of Chemistry, Youngstown State University, Youngstown, Ohio 44555, United States
*
S Supporting Information

ABSTRACT: Stereospecific SN2 conversion of configurationally pure acetobromoglu-

cose (2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl bromide) to the corresponding β-D-
glucopyranosyl azide is a useful exercise in the advanced organic undergraduate teaching
laboratory. The procedure is safe and suitable for small-scale implementation, and firm
proof of the stereochemical change is obtained from 1H NMR coupling constants. The
exercise provides students with experience in using important chiral pool natural product derivatives, reaction analysis by TLC, as
well as careful product isolation, purification, and spectroscopic identification.
KEYWORDS: Upper-Division Undergraduate, Laboratory Instruction, Organic Chemistry, Hands-On Learning/Manipulatives,
Carbohydrates, Chirality/Optical Activity, Conformational Analysis, Mechanisms of Reactions, NMR Spectroscopy,
Thin Layer Chromatography

N ucleophilic substitution at sp3 carbon, the SN1 and SN2

reactions, are two of the most important functional group
interconversions discussed early in the typical introductory level
organic chemistry sequence.1 These processes are used to
introduce the consequences of how subtle changes in structure
are able to have a dramatic effect on the mechanistic pathway
followed by organic compounds en route to product(s).
Although the stereochemical outcomes of these reactions are
discussed in detail in lectures, inversion of configuration in SN2
and typically racemate formation in SN1, there are very few
laboratory experiments available that give easily attainable
evidence for these changes.2 Here we introduce a useful SN2
experiment from sugar chemistry that uses a typical nucleophile
(azide) and leaving group (bromide) introduced in the
undergraduate class. Evidence for inversion comes from vicinal
coupling constants in the 1H NMR spectra of the starting
material and product.
The easily synthesized,3 and commercially available,4 sugar
derivative 2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl bromide
(acetobromoglucose, 1,) is an important glycosyl donor
(typically reacting via the SN1 pathway) that has found wide
application in the field of glycoside synthesis. The use of good
nucleophiles such as azide leads to stereospecific SN2
displacement on bromide 1 to yield, as is highlighted here,
the inverted glycosyl azide (2), which is a structurally well-
defined5 and useful precursor to biologically important glycosyl
amides (Scheme 1). In the experiment described here, students
Scheme 1. A Stereospecific SN2 Displacement Showing the
Inversion at an sp3 Carbon
were able to obtain proof for the stereochemical change at C1
when bromide 1 converts to azide 2 by comparing proton−
proton coupling constants for H1 of each material and then
using the Karplus equation to relate these values to the torsion
angles around C1−C2 of the two compounds. As well as being
a useful illustration of the inversion process in the SN2 reaction,
the sugar-derived compounds used in this exercise have
structures and spectra that are conducive to the discussion of
topics such as conformation, configuration, and phenomena
such as diastereotopicity.
■ EXPERIMENT
The commercially available bromide 1 is a well-behaved
colorless solid that is formed from D-glucose in a two-step
sequence of peracetylation followed by reaction with HBr in
acetic acid.3 The 1H NMR spectrum of the compound is
obtained in CDCl3 solution and shows it to be only the
configurationally pure α-anomer, which is explained in terms of
the anomeric effect operating on the pyranose ring.6 The SN2
experiment begins by dissolving the bromide in a 5:1 mixture of
acetone and water and then adding an excess of sodium azide
and allowing the mixture to stir overnight at room temperature.
Although azides are sometimes considered to be unsafe, we
have never encountered any problems with sugar-derived azides
such as 2 when simple safety precautions are in place.
Because glycosides 1 and 2 do not contain a chromophore,
they are not visible under a UV lamp; they are easily visualized,
however, by dipping the TLC plate into a solution of dilute
H2SO4 in ethanol which, after heating with a heat gun or on a
hot plate, revealed the compounds as dark spots on the white
silica background. When TLC indicated completion of the
reaction, evaporation of the acetone and an aqueous workup
provided the glucose-derived azide as a crystalline solid in 30−

Published: May 3, 2012

© 2012 American Chemical Society and
Division of Chemical Education, Inc. 943 dx.doi.org/10.1021/ed200596p | J. Chem. Educ. 2012, 89, 943−945
Journal of Chemical Education
80% yield after crystallization from hot methanol or isopropyl
alcohol (details are available in the Supporting Information).
The aqueous layer from the extraction contains the excess
sodium azide, so collection of these solutions from students
provides a simple and safe way to collect the waste.
The experiment typically requires two 3-h lab sessions to
complete; the first lab is spent setting up the reaction and
collecting spectral data for the glycosyl bromide 1, and the
second lab session involves product isolation, crystallization,
and characterization by NMR, melting point, IR, and
polarimetry, if available. Discussion of proton−proton coupling
constants occurs during the prelab component of the exercise,
and students are tasked with calculating these values as part of
the postlab homework and then using the values in conjunction
with the Karplus equation to prove if a stereochemical change
has actually occurred.
Obtained spectra offer opportunities to discuss both the
chemical and stereochemical changes occurring in this system;
in particular, the proton spectra of both compounds 1 and 2 are
exceptionally well-resolved thus allowing for convenient
coupling constant analysis. The signal for H1 in bromide 1
shows as a doublet at 6.61 ppm and the measured vicinal H1−
H2 coupling constant is found to be 4.0 Hz (Figure 1). This is
Figure 1. Partial proton spectrum of glycosyl bromide 1.
typical for the α-anomer of a D-glucopyranosyl system when the
pyranose ring is in the 4C1 chair conformation. The proton
spectrum of glycosyl azide product 2 reveals that the signal for
H1 has moved upfield (to 4.6 ppm) compared to the bromide
and that the H1−H2 coupling constant is now 8.8 Hz (Figure
2). This value is typical for the β-anomer of a D-glucopyranosyl
system in which the pyranose ring remains in the 4C1 chair
conformation.7 Overall, this change in coupling constants
serves as proof of a configurational change at C1 that results in
H1 and H2 now being aligned in an approximately anti
Figure 2. Partial proton spectrum of glycosyl azide 2.
944
Laboratory Experiment
relationship with the newly introduced azide group being in the
equatorial position. This change is readily discussed in terms of
chair conformations and Newman depictions (Figure 3).
Figure 3. Change in H1−H2 torsion angle upon stereochemical
inversion at C1.
■ HAZARDS
Because sodium azide is potentially explosive and is also toxic,
this should be noted in the prelab discussion. Students are
advised to wear gloves (nitrile or dish washing work well) while
handling this substance throughout the experiment as well as
observing the usual precautions of wearing safety glasses and a
lab coat or apron. We have used sodium azide on many
different scales in our teaching and research laboratories over
the past 20 years, in both carbohydrate and noncarbohydrate
experiments, and have never observed an explosion or an
adverse health effect. As long as students are aware of the
potential hazards of sodium azide, they treat it with due caution
and it becomes straightforward to handle as a nonvolatile solid.
Because an excess of sodium azide is used to enhance the rate
of the SN2 process, the extra salt must be recovered to ensure
safe disposal. This is readily achieved by collecting each of the
aqueous layers that students produce during the extraction
process, these solutions being composed of sodium azide and
the sodium bromide byproduct.
The use of dichloromethane in extractions may be
problematic, as it is considered to be a potential carcinogen.
Also, there have been several reports of explosions when
sodium azide and dichloromethane have been used together on
large scales.8 With these concerns in mind, ethyl acetate should
be used as a convenient low-boiling solvent for extractions in
this experiment. Acetone is highly flammable, irritating to the
eyes, repeated exposure may cause skin cracking or dryness and
vapors may cause drowsiness and dizziness. Methanol is
extremely flammable and toxic by inhalation and may be fatal
or cause blindness if swallowed. Concentrated sulfuric acid is
corrosive. Contact can cause severe damage to skin and eyes.
■
DISCUSSION
This experiment has been used for several years in the advanced
organic synthesis laboratory (typically 10−12 advanced under-
graduate students or beginning M.S. students) and familiarizes
students with important techniques and spectroscopic analyses
used in modern organic chemistry. The experiment typically
has been carried out on a 6−7 mmol scale; however, smaller
amounts are also workable, and yields are typically in the 30−
80% range, depending upon the skill level of the individual
student. Although the application has mostly been imple-
mented at the upper level with undergraduate students, and not
tested at the introductory level, this exercise should also be of
interest to those instructors running well-supervised laborato-
ries at the introductory level where the SN2 reaction is first
dx.doi.org/10.1021/ed200596p | J. Chem. Educ. 2012, 89, 943−945
Journal of Chemical Education Laboratory Experiment

discussed. With access to detailed NMR data (either from

student samples or the spectra available in the Supporting
Information), it is possible to discuss proton assignments and
coupling constants, as well as other aspects of the spectra. For
example, the diastereotopicity of the C6 protons is readily
apparent in azide 2 (H6 and H6′ are both observed as doublets
of doublets, and H5 is a resolved doublet of doublet of
doublets), and the use of COSY spectra for complete
assignment of the ring proton signals for azide 2 is an
interesting application as H2, H3, and H4 each appear in the
1
H spectrum as apparent triplets (or doublets of doublets) with
approximately equal coupling constants.

■ CONCLUSION
This report introduces a useful SN2 laboratory experiment that
allows students to observe stereochemical inversion at C1 of a
D-glucopyranosyl system. While gaining experience in the
synthesis of important sugar derivatives, the exercise provides
opportunities for students to use coupling constant data, the
Karplus equation, and conformational analysis to establish that
a stereochemical inversion has occurred.

■ ASSOCIATED CONTENT
* Supporting Information
S

Experimental procedure; instructor’s notes (including safety

information); 1H, 13C, and COSY NMR spectra for compounds
1 and 2. This material is available via the Internet at http://
pubs.acs.org.

■ AUTHOR INFORMATION
Corresponding Author
*E-mail: (J.A.J.) jajackson@ysu.edu; (P.N.) pnorris@ysu.edu.
Notes
The authors declare no competing financial interest.

■ REFERENCES
(1) Carey, F. A.; Giuliano, R. M. Organic Chemistry, 8th ed.; McGraw
Hill Higher Education: New York, 2010; pp 137−183.
(2) For a recent laboratory experiment based on the SN2 reaction at a
primary carbon see: Esteb, J. J.; Magers, J. R.; McNulty, L.; Morgan,
P.; Wilson, A. M. J. Chem. Educ. 2009, 86, 850−852. For a recent
example of a laboratory exercise in which inversion is observed see:
Van Draanen, N. A.; Hengst, S. J. Chem. Educ. 2010, 87, 623−624.
(3) Fischer, E. Ber. Dtsch. Chem. Ges. 1916, 49, 584−585.
(4) Acetobromoglucose is available commercially in multigram
batches from companies such as Sigma-Aldrich (catalog number
A1750).
(5) Temelkoff, D. P.; Norris, P.; Zeller, M. Acta Crystallogr., Sect. E:
Struct. Rep. Online 2004, E60, o1975−o1976.
(6) Nisic, F.; Andreini, M.; Bernardi, A. Eur. J. Org. Chem. 2009, 33,
5744−5751.
(7) The spectra obtained for student samples of azide 2 match those
reported previously for this compound; Orth, R.; Pitscheider, M.;
Sieber, S. A. Synthesis 2010, 2201−2206.
(8) Conrow, R. E.; Dean, W. D. Org. Process Res. Dev. 2008, 12,
1285−1286.

945 dx.doi.org/10.1021/ed200596p | J. Chem. Educ. 2012, 89, 943−945