Louise Quelch

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Justification
Major changes were made in almost every section of the report. Adding a title
which fully described the paper, but was still able to be concise was very
important. This is because it is the first thing that the readers will see and will
determine whether they will continue reading [1]. Usually read second, an abstract
that summarise the whole report while being able to stand on its own and hold
the readers attention it is vital [2]. Pointed out by both reviews, the introduction,
had little to no background or context. However, now there is crucial information
such as detail about the different anomers and the reaction mechanism. The
introduction was structured with a broad start and gradually got to the specific
aims of the investigation [3]. The results were displayed logically from most
important to the justification in the discussion, 1H NMR, to least important, the
yield. The discussion had a lot of work done on the structure and the content. It
was structured with information that most supported the conclusions at the start,
and as it went on the information became more general [4]. The paragraph
justifying the assignment of the hydrogens was removed. For much of the
audience this information would be irrelevant and it was not highly relevant in
coming to the conclusions. There was also many general improvements. One of
these was catering the paper for an audience with a minimal chemical
background. This resulted in many ideas and terms being defined and explained
in depth. Another improvement was the visual appeal of the figures and how
they were integrated into the text. These improvements outlined and many
others demonstrate how the reviewed paper is more effective at communicating
the science to the desired audience.

References
[1] T. M. Annesley, Clinical Chemistry, 56:3, 257 -260
[2] T. M. Annesley, Clinical Chemistry, 56:4, 531 -524
[3] T. M. Annesley, Clinical Chemistry, 56:5, 708713
[4] T. M. Annesley, Clinical Chemistry, 56:11, 16711674

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Anomeric composition of D-glucose

pentaacetate after synthesis
Louise Quelch

Abstract
In the formation of ethers as part of a cyclic ring, a new chiral centre is formed.
This causes two anomers of the same molecule. and -D-glucose pentaacetate
are an example of this. The aim of this investigation was to synthesis D-glucose
pentaacetate and determine the anomeric composition of the purified product.
This was achieved by reacting -D-glucose with anhydrous sodium acetate and
purifying the precipitate. A 1H NMR was run on this product and melting point of
108.5 123.2 C recorded. The 1H NMR was compared to 1H NMR spectrums of
known and -D-glucose pentaacetate samples (provided by The University of
Queenslands SCMB). From the comparison it was clear that the product had
aspects of both anomers. The intergrals of the peaks demonstrate that there was
significantly more of the anomer present. Another way to identify the anomer
is through the differing coupling constants cause by dihydrogen interaction
between the hydrogen atoms on and adjacent to the anomeric carbon, either the
diaxial, 7 - 10 Hz, or axial-equatorial, 2 6 Hz. The coupling constant
determined was 10.88 Hz. While this does not fit directly into either range it is
clearly closer to the expected coupling constant for the anomer. For these
reasons it was determined that the major product was the anomer.

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Introduction
Anomers have different chemical and physical properties, so it is vital to be able
to determine procedures to form each anomer. For example -D-glucose
pentaacetate (figure 2) has been shown to induce insulin release in rate,
however its anomer -D-glucose pentaacetate (figure 1) did not. This is just one
situation where it is vital to be able to distinguish between the two anomers. [1]

Figure 1: -D-glucose-pentaacetale

Figure 2: -D-glucose pentaacetale

D-glucose pentaacetate (figures 1 and 2) is formed through the acetylation of D-glucose (figure 3). Acetylation is the process of adding an acetyl group (RCOCH3) to a hydroxyl group (R-OH). In the case of glucose there are 5 hydroxyl
groups, which are all acetylated.

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Figure 3: Acetylation of D-glucose; R is the alcohol groups on the glucose

- D-glucose pentaacetate and -D-glucose pentaacetate differ in the orientation

of the functional group on C-1 (position 1, figure 1) the anomer has the group
in the axial position (perpendicular to the plane of the molecule), whereas the
anomer has the group in the equatorial position (in the plane of the molecule).
Which anomer forms depends on the direction that alcohol group and the
aldehyde interact to for a hemiacetal. Once the 6 membered ring is formed these
anomers are locked. Only through the opening and reclosing of the 6 membered
ring, mutarotation [2], can D-glucose pentaacetate (and glucose) alteration
between the and anomers.
The aim of this experiment was to synthesis glucose pentaacetate through the
acetylation of -D-glucose. Once synthesised and purified the 1H NMR was used
to determine the anomeric composition, ie whether the product was -Glucose
pentaacetate or -Glucose pentaacetate.

Method
Formation of D-glucose pentaacetate:
-D-glucose (5.0007 g, 0.027757 mol) was added to anhydrous sodium acetate
(2.51 g, 0.0306 mol) and refluxed for an hour with acetic anhydride (25 mL,
0.264 mol). The solution was added to distilled ice water for 15 minutes. The
solution was vacuum filtered and the precipitate collected. This product was then
recrystallised by dissolving in minimal boiling methanol and allowed to crystallise
again. The product was collected and rinsed with cold methanol.
1

H NMR Procedure:

The product (0.3 g) was dissolved in deuterochloroform (1 mL). A 1H NMR

spectra (400 Hz) was run on this solution.

Results
The 1H NMR results are displayed in table 1. The NMR shows 11 major peaks
many of which have been split into sub-peaks due to influence of neighbouring
hydrogens, this is described by the multiplicity. Each peak can represent multiple
hydrogens, this was determined by integration of the major peak. Another
conclusion of the NMR is the coupling constant. It is calculated by the difference
in chemical shift of the lowest and highest sub-peaks multiplied by the frequency
(400 Hz). For raw data refer to Appendix 1.

Chemical
shift

No. of
Hydrogens

Multiplicity

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Coupling
constant

Assignment

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average
(ppm)
5.7146
5.2527
5.1285

1
1
2

4.289

4.11125

3.83775

2.1141
2.0856
2.0325
2.0126

3
3
6
3

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(Hz)
Doublet
Triplet
Doublet of
triplets
Doublet of
doublets
Doublet of
doublets
Doublet of
doublets of
doublets
Singlet
Singlet
Singlet
Singlet

10.88
24.88
27.84

1
2
3,4

22.72

19.64
22.2

0
0
0
0

7
11
8 and 10
9

Table 1 - 1H NMR results (Note: assignment numbers refer to Figure 1)

The mass of the final purified glucose pentaacetate was 7.37 g, giving a yield of
68.0 % and a melting point of 108.5 123.2 C.

Discussion
In this study D-glucose pentaacetate was synthesised, it was determined that the
resulting product was a mixture of both the and anomers, with -D-glucose
pentaacetate being the predominant species. This finding is supported by both
the 1H NMR and the melting point range.
When compared to 1H NMR spectrums -D-glucose pentaacetate and -D-glucose
pentaacetate (see appendix 2and 3), provided by The University of Queenslands
SCMB, the spectrum of the product could clearly been seen to align more
strongly with that of the anomer. One example of this is the peak
corresponding to the hydrogen attached to C-1 (position 1, figure 1). This
hydrogen is the least shielded by electrons, since there is a greater number of
electron withdrawing oxygen atoms in close proximity. This causes the peak to
be lower field ie to have a higher chemical shift, which in the case of the
hydrogen on C-1 in the anomer is approximately 5.7 ppm. However in the
anomer it is around 6.3 ppm. When the product was tested, peaks at both 5.7
and 6.3 ppm were observed. Appendix 1, page 6, shows this. From the relative
integrals of these peaks it can be seen that the amount of anomer in the final
product is significantly larger.
Another way to differentiate between the two anomers is the value of the
coupling constant of in the peak from the hydrogen bonded to C-1. As seen in
figures 1 and 2, -D-glucose pentaacetal and -D-glucose pentaacetal have the
substituents in the equatorial and axial positions respectively. This means that in
the anomer the Hydrogen in position 1 is axial. This causes diaxial interactions
between itself and the hydrogen in position 2. This causes coupling constants of
7 10 Hz. Conversely the hydrogen in position 1 in the isomer is equatorial
causing equatorial-axial interactions, resulting in coupling constants of 2 - 6 Hz.
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In table 1 it can be seen that the coupling constant recorded is 10.88 Hz. While
this value does not fit directly in the diaxial interactions range it is significantly
closer and therefore it can be concluded that the majority of the product is -Dglucose pentaacetate.
More evidence to support the explaination of both anomer existing in the product
it the recorded melting range. The literature values for the melting points of Glucose pentaacetate and -Glucose pentaacetate are 131 132 C [3] and 111
112 C[4] respectively. The large range of the melting point, 108.5 123.2 C, can
be attributed to the final product containing the and anomers. The melting
point recorded does not seem very reliable due to its low start point, compared
to the literature values of both anomers. One likely explaination is that the
product was not completely dry, ie still containing methanol, when the melting
point was recorded.
As seen in inaccuracy of the melting point, this study has many possible ways in
which it could be improved. Another example of this is in the low yield of 68.0 %.
One literature value for the yield of a very similar procedure was 85 % [5]. The
calculated yield was decrease due to loss of both products and reactants
throughout the reaction. Many reactants were lost through the reflux, as not all
of the solid was dissolved into solution. In situations where time is not limited it
would improve the yield to leave the mixture to reflux longer.

Conclusion
The major product produced was -Glucose pentaacetate. This was determined
through 1H NMR, by using relationships with axial and equatorial hydrogens and
analysing the 1H NMR of know sample of and -D-Glucose pentaacetate. This
investigation will be beneficial for any future need to determine different
procedures for the synthesis of the or - D-glucose pentaacetate.

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Appendix 1: Product 1H NMR

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Appendix 2: -D-glucose pentaacetate 1H NMR

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Appendix 3: -D-glucosepentaacetate 1H NMR

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References
[1] W. J. Malaisse, H. Jijakli, M. M. Kadiata, A. Sener, O. Kirk, Biochemical and
Biophysical Research Communications, 1997, 231, 435 - 436
[2] A. M. Silva, E. C. da Silva, C. O. da Silva, Carbohydrate Research, 2006, 341,
1029 1040
[3] U. Niedballa, H. Vorbruggen, J. Org. Chem., 1974, 39, 3659
[4] F. Dasgupta, P. P. Singh, H. C. Smvsrava, Ekevier Scientific Publishing
Company, 1980, 80, 346
[5] A. Pulsipher and M. N. Yousaf Chem. Commun., 2011, 47, 523

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