Cooperative learning in science education is addressed in this article. How students use a very relevant topic of anti-cancer agents, and the novel technique of (Heteronuclear single Quantum Correllation Spectroscopy )2D -HSQC FT-NMR to organize spectra data is shown. Here, undergraduates become familiar with making plots of 1H FT-NMR and 13C FT-NMR , learning FT-NMR data processing (spinworks) and also use Chemdraw NMR to present data take with a Varian 600 MHz FT-NMR spectrometer.

1. Cooperative learning using FT-NMR :
Students: Mohammed Izmikna, Kasandra Dorce, Mohammed
Sherwani, Samira Izmikna,
By Dr. Robert Craig, Ph.D.
Cooperative learning in science education is addressed in this
article. How students use a very relevant topic of anti-cancer
agents, and the novel technique of (Heteronuclear single
Quantum Correllation Spectroscopy )2D -HSQC FT-NMR to
organize spectra data is shown. Here, undergraduates become
familiar with making plots of 1H FT-NMR and 13C FT-NMR ,
learning FT-NMR data processing (spinworks) and also use
Chemdraw NMR to present data take with a Varian 600 MHz
FT-NMR spectrometer.We can make a tentative spectulation
that For the side chain methyl ester, the coupling constant JH2-
H3 changes from 2 Hz to 5 Hz. This confirms a shift in
conformational equilibria, after addition of this group. The
“deshielding effect” of a carboxylate group on the monomer of
curcumin is shown by 2D -HSQC FT-NMR data and the
ChemdrawNMR software.
Cooperative learning using FT-NMR :
Abstract

2. Cooperative learning in science education is addressed in this
article. How students use a very relevant topic of anti-cancer
agents, and the novel technique of HSQC FT-NMR to organize
spectra data is shown. Here, undergraduates become familiar
with making plots of 1H NMR and 13C NMR , learning FT-NMR
data processing (spinworks) and also use Chemdraw NMR to
present data take with a Varian 600 MHz FT-NMR
spectrometer. For the side chain methyl ester, the coupling
constant JH2-H3 changes from 2 Hz to 5 Hz. This confirms a
shift in conformational equilibria.
• The “deshielding effect” of a carboxylate monomer of
curcumin is shown by 2D -HSQC FT-NMR data and the
ChemdrawNMR software.
In this paper, student projects are given as an example on how
to introduce FT –NMR into the undergraduate curriculum.
The deshielding of the antimitotic agent curcumin and it side
chain methyl ester have been studied by Nmr spectroscopy
and by molecular modeling using ChemDrawNMR .
Upon carboxylation, some proton shifts change. Students
proficient in ChemDrawNMR show this in their analysis.

3. For the side chain methyl ester, the coupling constant JH2-H3
changes from 2 Hz to 5 Hz. This confirms a shift in
conformational equilibria.
We will incorporate NMR experiments that illustrate the
application of high resolution NMR spectroscopy to the
structure determination of Anti-Cancer agents.
Adding NMR spectroscopy to a students repertoire of skills will
greatly enhance the laboratory learning experience and enable
us to adopt experiments that have been successful elsewhere
at utilizing NMR spectroscopy as an essential teaching tool.
This paper will also examine the benefits of students involve in
performing NMR experiments at the undergraduate level
As far as Curcumin is concerned, The effect of the carboxyl
group on the proton spectra(deshielding effect) is clearly shown
with chemdrawNMR software platform.
current guidelines of the Committee on Professional Training1
(CPT) of the American Chemical Society (ACS) are very clear on

4. its expectation for inclusion of instrumentation, and in
particular NMR spectroscopy, in the chemistry curriculum. An
excerpt from this document states, “A department should have
several major pieces of sophisticated equipment suitable for
undergraduate instruction as well as for research. One of these
must be an NMR spectrometer.”1 Later in this same document
it is noted that the instrument should be an FT-NMR. In
addition, the most current proposed revision to this document
states, “The laboratory experience should include synthesis of
molecules, measurement of chemical properties and
phenomena, “hands-on” experience with modern
instrumentation, applications to real-world problems, and
computational data analysis and modeling.”2

5. Curcumin’s broad spectrum of anti-oxidant, anti-carcinogenic,
anti-mutagenic, and anti-inflammatory properties makes it
particularly interesting for the development of pharmaceutical
compounds. Due to curcumin’s various effects on the function
of numerous unrelated membrane proteins, it has been
suggested that it affects the properties of the bilayer itself.
Despite intense interest in the physiological effects of
curcumin, a general mechanism for its action has not been
identified.
Figure 1
Keto-enol form of curcumin, the dominant tatuomer of
curcumin. The keto-enol from is stabilized by an intramolecular
hydrogen bond, shown here by a dashed red line.
Here is a mechanism of interest to many.

6. MATERIALS AND METHODS
Curcumin (>94%) was purchased from Sigma and used without
further purification
This red powder was successively subjected to 1H NMR, 13C
NMR, and 2D -HSQC FT-NMR analysis for structure
For the Varian 600mHz, 5mm NMR Sample tubes were used
from NewEra, inc

7. . The NMR sample tubes were“L” Series 5mm NMR tubes
(4.960 ± 0.006mm OD; 0.40mm nominal wall; 0.0025mm
roundness).
All spectra was processed from the Varian using spinworks
platform.The specta was subsequently confirmed using
Chemdraw NMR. It was convinent to use Spinworks to analyze
spectra. The Spinworks software, created by Kirk Marat. also
provides us with excellent ppm shifts for both spectra.
FT NMR
All of the experiments were performed on a Varian Infinity 600
MHz solid-state NMR spectrometer. Each sample was
equilibrated for at least 30 minutes before starting the
experiment. 2D 13C-1H correlated (HECTOR) NMR spectra were
obtained using a spin-echo pulse sequence (90°-τ–180°-τ-
acquisition; τ = 125 μs) with a 90° pulse length of 5 μs under a
30 kHz continuous-wave proton decoupling.

8. Chemical shifts were referenced by setting the isotropic
chemical shift peak of TMS to 0 ppm. 2D 13C-1H correlated
(HECTOR) quadrupole coupling spectra were recorded using a
quadrupolar echo pulse sequence (90°-τ–90°-τ-Acquisition; τ =
80 μs) without proton decoupling.
RESULTS AND DISCUSSION
Since curcumin affects such a large array of unrelated
membrane proteins at approximately similar concentrations, it
has been proposed that curcumin can regulate the action of
membrane proteins indirectly by changing the physical
properties of the membrane rather than by the direct binding
of curcumin to the protein.
High resolution 13 C and 1H NMR , 2D 13C-1H correlated
(HECTOR), and 2D 1H-1H correlated (COSY) spectroscopy
techniques will be used for elucidating skeletal arrangement of
monomer units.
Applications that also use the 2D 1H-13C HSQC experiment are
gaining more interest as a result of the growing feasibility of
acquiring these spectra routinely. The 2D HSQC experiment

9. contains additional information (i.e. 13C chemical shift) as well
as easier identification of labile and diastereotopic protons
2D NMR specra may be obtained that indicate coupling
between hydrogens and carbons to which they are attached. In
this case it is called heteronuclear correlation spectroscopy
(HECTOR, HSQC, or C-H HECTOR).
When ambiguities are present in one-dimensional 1H and 13C
NMR spectra, a HECTOR or HSQC spectrum can be very useful
for assigning preciscely which hydrogens and carbons are
producing their respective peaks.
In a HSQC spectrum a 13 C spectrum is presented along one
axis and a 1H spectrum is shown along the other. Cross peaks
relating the two types in a HSC spectrum indicate which
hydrogens are attached to which carbons in a molecule, or vice
versa.
These cross peaks correlations are the result of instrumental
parameters specified on the NMR spectrometer. If imaginary

10. lines are drawn from a given cross peak in the x-y field to each
respective axis,
The cross peak indicates to the hydrogen giving rise to the
corresponding 1H NMr signal on one axis and is coupled or
attached to the carbon that gives rise to corresponding 13C
NMR signal on the other axis.
Thus, it is readily apparent which hydorgens are attached to
which carbons
FIG 1:The effect of the carboxylated curcumin on proton signal

12. The effect of the carboxylated curcumin on proton signal
Referring to the chemdrawNMR data below
curcumin from
proton chemdraw
shift atom index coupling partner constant and vector
5.35 7 delta (ppm)
5.35 25
7.16 24
20 20 1.5 H-C*C*C*-H
6.99 21
20 7.5 H-C*C*-H
7.16 6
20 20 1.5 H-C*C*C*-H
6.79
21 21 7.5 H-C*C*-H
24 24 1.5 H-C*C*C*-H
6.99 3

13. 4 4 7.5 H-C*C*-H
6.79 4
3 3 7.5 H-C*C*-H
6 6 1.5 H-C*C*C*-H
3.83 10
3.83 27
4.59 13
7.6 28
30 15.1 H>C=C<H
7.6 29
31 15.1 H>C=C<H
6.91 30
28 15.1 H>C=C<H
6.91 31
29 15.1 H>C=C<H
curcumin from
proton chemdraw
carboxylated
shift atom index coupling partner constant and vector
5.35 25
11 35
7.16 24
20 20 1.5 H-C*C*C*-H
6.99 21
H-C*C*-
20 7.5 H
7.3 @@ 6
4 4 1.5 H-C*C*C*-H
6.79
H-C*C*-
21 21 7.5 H
24 24 1.5 H-C*C*C*-H
7.13 3
H-C*C*-
4 4 7.5 H
7.26 '@@ 4
H-C*C*-
3 3 7.5 H
6 6 1.5 H-C*C*C*-H
3.83 10
3.83 27
2.3 29

14. 31 31 7.1 H-CH-CH-H
2.3 32
31 31 7.1 H-CH-CH-H
4.59 13
2.01 31
29 29 7.1 H-CH-CH-H
7.6 36
38 38 15.1 H>C=C<H
6.91 38
36 36 15.1 H>C=C<H
6.91 39
37 37 15.1 H>C=C<H
•
above is Figure 2:13C spectra of Curcumin

15. above is Figure 3: the 2D1H -13C HSQC spectra of Curcumin
• Let’s dive right in, as the research students have provided
the spectra and determine the HSQC for Curcumin, with
the aid of the ChemdrawNMR software, and previous scan
of curcumin (proton and 13C). It is beneficial to keep
these spectra on hand. The Spinworks software, created
by Kirk Marat.also provides us with excellent ppm shifts
for both spectra
Working from top down, and left to right, the HSQC for
curcumin reads as such. The first peak evident in the
spectra is 13C at 55.934 ppm, And crossed with

16. Proton(designed 14) at 3.9620 ppm. The next peak is with
Proton(designed 12) at 5.8592 and a 13 C at 101 ppm
• This hydrogen must be attached to the OH group , or
might be the hydrogen in between the carbonyls on the
hexadione bridge.
• The carbon 13 peak at 109.3 cross with several protons.
Referenced with the spinworks data table for curcumin
proton data taken the Varian 600 MHz we have for Peak 3
in the HSQC specta with Peak 12 at 5.8592 ppm And
Peak 13 at 5.8124 ppm in the proton spectra. Please refer
to table one for the spinworks data.
• The carbon 13 peak at 109.3 ppm coupled with a hydrogen
(peak 5 at 7.1427 ppm) is an aromatic hydrogen. This
hydrogen resides on a benzene ring, and is obviously
confirmed by coupling with an aromatic 13C at 109.3 ppm
• It is this hydrogen that will be effected in the
carboxyalated form of curcumin. The carbon peaks at
122.6 and 123.8 ppm with peak 3 and 4 (off diagonal) of
the proton data gives some modest peaks in the
specta.Also evident are Peak 3 (122.6 ppm 13C with (6.473
ppm 1H, 6.503 ppm 1H )And, With peak 7 and 8 (shown in
the off diagonal) coupling 123.8 ppm 13C with (6.473 ppm
1
H , 6.503 ppm 1H). These hydrogens are on aromatic ring
next to hydroxyl groups. A Carbon of 114 ppm is

17. appropriate to be adjacent to these hydrogens. As
reference by the ChemdrawNMR softwareplatfom”The
benzene CH of which there are 3, give rise to 7.16 ppm,
6.99 ppm and 6.79 ppm. “On the hexadienone bridge,
between the two benzene rings (aromatic rings) are 2 pairs
of equivalent protons, (see table 2). The software also
allows for shift corrections
• It is this hydrogen that will be effected in the
carboxyalated form of curcumin. The carbon peaks at
122.6 and 123.8 ppm with peak 3 and 4 (off diagonal) of
the proton data gives some modest peaks in the
specta.Also evident are Peak 3 (122.6 ppm 13C with (6.473
ppm 1H, 6.503 ppm 1H )And, With peak 7 and 8 (shown in
the off diagonal) coupling 123.8 ppm 13C with (6.473 ppm
1
H , 6.503 ppm 1H). These hydrogens are on aromatic ring
next to hydroxyl groups. A Carbon of 114 ppm is
appropriate to be adjacent to these hydrogens. As
reference by the ChemdrawNMR softwareplatfom”The
benzene CH of which there are 3, give rise to 7.16 ppm,
6.99 ppm and 6.79 ppm. “On the hexadienone bridge,
between the two benzene rings (aromatic rings) are 2 pairs
of equivalent protons, (see table 2). The software also
allows for shift corrections
Mono Carboxyated is here

18. Figure 4, 5 and 6 are the 2D1H -13C HSQC spectra of mono
carboxylated Curcumin
Assign ment of theMONO-CARBOXYLATED CURCUMIN HSQC-ON THE
WALL

19. The peak at 6.953 ppm for hydrogen (and 6.94 ppm) corresponds to a
C=C-OH on the right benzene ring of the mono carboxylated form.
The carbon associated with this Hydrogen is the signal at 114 ppm. The
carbon experiencing a more electronegative environment is next to this
Carbon at 123.8 ppm . this carbon at 123.8 ppm gives a signal with a
proton at 7.063 ppm. This 7.063 also spin couples with a carbon
At 109 ppm. This is associated with a similar fragment,C=C-OCH3 on
the top of rightsided benzene ring. The carbon at 109 ppm couples
twice
With protons at 7.129 ppm and the one just mentioned thereafter.
The proton at 7.143 ppm we can designate Ha , which resides on top of
the rightsided benzene ring couples with 3 carbons at 109 ppm, 111.8
ppm
And 122.0 ppm. This fragment is the “Ha-‘C=C-OCH3”.
The signal at 7.15 ppm we can designate Hb , which resides on top of
the rightsided benzene ring, as well, and couples with 3 carbons at 109
ppm,
111.8 ppm And 122.0 ppm. This fragment is the “Hb-‘C=C-OCH3”.
a C-C=C-OH makes up the final piece in this portion of the spectra. It is
the bottom of the benzene ring on the right side
of the carboylated curcumin, unaffected by this substituent group. The
Hc-C-C=C-OH is responsible for the peak at 7.19 ppm,120ppm

20. and 7.19 ppm,120 ppm
This cluster of peaks resides in the bottom right portion of the HSQC spectra
mono carboxylated curcumin molecule.
The carbon at 140.5 ppm (Ca)signals with a hydrogen residing on the hexadione
brigde (H-Ca=C=H)
The carbon at 139.5 ppm (Cb)does the same for this fragment (H-C=Cb=H)
(5)Figure 4, 5 and 6 are the 2D1H -13C HSQC spectra of mono
carboxylated Curcumin

21. U(6)
The one carbon peak at 5.35 ppm sweeps the proton signals with 3.855 ppm and 3.82 ppm.
This is the C-O-CH3 group
conclusion
Heteronuclear Single Quantum Coherence (HSQC)Plots 1H NMR on x-axis and
13
C NMR on y-axis and Utilizes 1 bond coupling between H and C, Eliminating all
the H containing C’s eliminates many C’s assignments. Leaves only on-H
containing C’s to assign.
Using information from 1H NMR data alone is not a new concept. However, applications that
also use the 2D 1H-13C HSQC experiment are gaining more interest as a result of the growing
feasibility of acquiring these spectra routinely. The 2D HSQC experiment contains additional
information (i.e. 13C chemical shift) as well as easier identification of labile and diastereotopic

22. protons. I would like to thank the students and staff at the college of Staten Island, CUNY for
making this work possible. I find cooperative learning to be very important because it is crucial
for our students to learn to work in groups. This not only helps develop their social skills, but
also enhances their ability to develop the skills necessary to work collaboratively when they
enter graduate school.
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Educational Research Association”, Chicago, IL, April 3-7, 1991. [revised, 20 January,
1996]
• Implementation of FT-NMR Across the Chemistry Curriculum , Committee on
Professional Training (CPT) of the American Chemical Society, CCCE Dunal Admin –
October 16, 2009 to October 16, 2009.
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concepts in the context of solution calorimetry.” (2003). International Journal
of Science Education, 25(7), 779-800.
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curcumin
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