Structure Elucidation with Nuclear

Magnetic Resonance (NMR)

This was prepared from any material downloaded from internet.

NMR Applications in Chemistry

Chem. Shift
J Coupling

NMR
spectrum

Peak Int.

Structure

NOE

10
6'

CH3

5'

CH2OH
OO

Correlation

H-HC-H

4'

5''

OH 1'
3' 2'

6''

COOCH2
7''

2''

Applications:
Sample quality control for Synthetic works.
Elucidation of chemical structures.
Getting functional group, bonding, dynamics, kinetics and
chemical exchange information of molecules.
3D structures of the molecules.

3
4

OH
1''

3''

H 7

OH

4''

O
1

O
OH

Some common NMR experiments:

Connections through bondsspin-spin coupling
Homonuclear 1D2D 1H-1H COSY, DQF-COSY, TOCSY
usagespin system assignment
Heteronuclear
Direct (detect 13C): APT, DEPT, HETCOR
Inverse (detect1H): HMQC, HSQC, HMBC, HSQC-COSY,
HSQC-TOCSY, HMQC-TOCSY
usageassigning heteronucleiconnecting spin systems
Connections through spacedipolar coupling
1D2D NOESYROESY, HOESY(HSQC-NOESY)
usage connecting spin systems structure determination

2D COSY Homonuclear Shift COrrelation SpectroscopY

x

90
t1

t2
AQ

The basic COSY (x=45 or 90) pulse sequence

Vicinal

Geminal

H H
C C H
COSY spectrum is used for determining the connectivities
between protons on the basis of geminal and vicinal couplings.
Disadvantage: bulky dispersive diagonal peaks.

2D Gradient COSY-45
1

2 3

Current Data Parameters

NAME
butcosy
EXPNO
1
PROCNO
1

ppm

3-4

2 3

1.0

2-3
1-2
1.5

2.0

H1 H2 H3 H4
HO C C C C H

2.5

H H H H

3.0

2.5

2.0

1.5

1.0

============ CHANNEL f1
NUC1
1H
P0
3.00
P1
6.00
PL1
-4.00
SFO1
500.1310815

=============
usec
usec
dB
MHz

============ GRADIENT CHANNEL ========

GPNAM1
sine.100
GPNAM2
sine.100
GPX1
0.00 %
GPX2
0.00 %
GPY1
0.00 %
GPY2
0.00 %
GPZ1
10.00 %
GPZ2
10.00 %
P16
1000.00 usec
F1 - Acquisition parameters
ND0
1
TD
256
SFO1
500.1311 MHz
FIDRES
8.535453 Hz
SW
4.369 ppm
FnMODE
undefined

3.0

F2 - Processing parameters
SI
2048
SF
500.1300144 MHz
WDW
QSINE
SSB
0
LB
0.00 Hz
GB
0
PC
0.20

3.5

F1 - Processing parameters
SI
1024
MC2
QF
SF
500.1300144 MHz
WDW
QSINE
SSB
0
LB
0.00 Hz
GB
0

3.5

F2 - Acquisition Parameters
Date_
20001102
Time
8.04
INSTRUM
DRX500
PROBHD
5 mm TBI 1H/
PULPROG
cosygp
TD
1024
SOLVENT
CDCl3
NS
1
DS
16
SWH
2185.315 Hz
FIDRES
2.134096 Hz
AQ
0.2345700 sec
RG
40.3
DW
228.800 usec
DE
6.00 usec
TE
288.0 K
D0
0.00000300 sec
D1
1.60000002 sec
D13
0.00000300 sec
D16
0.00010000 sec
IN0
0.00045765 sec

ppm

2D DQFCOSY(Double-Quantum Filtered COSY)

90

90
t1

90
t2

) allow J coupling constant measurements

) observing cross-peaks close to diagonal
) But lower sensitivity (~40%) of COSY expt.

2D Gradient DQF-COSY
Current Data Parameters
NAME
gbc35dqf
EXPNO
2
PROCNO
1

ppm
3.6
3.8
4.0
4.2
4.4
4.6
4.8
5.0
5.2
5.4
5.6
5.8
6.0
6.2

6.2

6.0

5.8

5.6

5.4

5.2

5.0

4.8

4.6

4.4

4.2

4.0

3.8

3.6

ppm

F2 - Acquisition Parameters
Date_
20001103
Time
4.01
INSTRUM
DRX500
PROBHD
5 mm TBI 1H/
PULPROG
cosydfetgp.2
TD
4096
SOLVENT
Pyr
NS
8
DS
16
SWH
3822.630 Hz
FIDRES
0.933259 Hz
AQ
0.5359376 sec
RG
512
DW
130.800 usec
DE
6.00 usec
TE
288.0 K
D0
0.00000300 sec
D1
1.60000002 sec
D11
0.03000000 sec
D16
0.00010000 sec
DELTA
0.00110300 sec
DELTA1
0.00110800 sec
IN0
0.00026161 sec
l3
256
============ CHANNEL f1
NUC1
1H
P1
5.80
P2
11.60
PL1
-4.00
SFO1
500.1317609

=============
usec
usec
dB
MHz

============ GRADIENT CHANNEL ========

GPNAM1
sine.100
GPNAM2
sine.100
GPNAM3
sine.100
GPX1
0.00 %
GPX2
0.00 %
GPX3
0.00 %
GPY1
0.00 %
GPY2
0.00 %
GPY3
0.00 %
GPZ1
30.00 %
GPZ2
10.00 %
GPZ3
50.00 %
P16
1000.00 usec
F1 - Acquisition parameters
ND0
1
TD
512
SFO1
500.1318 MHz
FIDRES
7.465717 Hz
SW
7.643 ppm
FnMODE
undefined
F2 - Processing parameters
SI
4096
SF
500.1300038 MHz
WDW
SINE
SSB
2
LB
0.00 Hz
GB
0
PC
0.20
F1 - Processing parameters
SI
1024
MC2
echo-antiecho
SF
500.1300038 MHz
WDW
SINE
SSB
2
LB
0.00 Hz
GB
0

3J=8.00Hz

TOCSY (TOtal Correlation SpectroscopY) or

HOHAHA(Homonuclear Hartman-Hahn Spectroscopy)
t1

mixing time
MLEV17

t2
AQ

Pulse sequence for a TOCSY spectrum.

H1 H2 H3 H4

H1 H2 H3 H4

H1 H2 H3 H4

HO C C C C H HO C C C C H HO C C C C H
H H H H

H H H H

H H H H

COSY

RL-COSY

TOCSY

Different mixing time gives different degree of relay of

correlation. At small mixing time, TOCSY spectrum is similar to
COSY spectrum. At long mixing time, gives total correlation.

2D Gradient TOCSY, mixing time=10ms

Current Data Parameters
NAME
buttoc10
EXPNO
1
PROCNO
1

2 3

ppm

3-4

2 3

1.0

2-3
1-2

1.5

2.0

H1 H2 H3 H4
HO C C C C H

2.5

H H H H
3.0

Same as COSY spectrum

3.0

2.5

2.0

1.5

1.0

============ CHANNEL f1
NUC1
1H
P1
5.60
P2
11.20
P5
19.33
P6
29.00
P7
58.00
P17
2500.00
PL1
-4.00
PL10
10.00
SFO1
500.1310815

=============
usec
usec
usec
usec
usec
usec
dB
dB
MHz

============ GRADIENT CHANNEL ========

GPNAM1
sine.100
GPNAM2
sine.100
GPX1
0.00 %
GPX2
0.00 %
GPY1
0.00 %
GPY2
0.00 %
GPZ1
30.00 %
GPZ2
30.00 %
P16
1000.00 usec
F1 - Acquisition parameters
ND0
1
TD
256
SFO1
500.1311 MHz
FIDRES
8.535453 Hz
SW
4.369 ppm
FnMODE
undefined
F2 - Processing parameters
SI
1024
SF
500.1300144 MHz
WDW
SINE
SSB
3
LB
0.00 Hz
GB
0
PC
0.20

3.5

3.5

F2 - Acquisition Parameters
Date_
20001102
Time
4.14
INSTRUM
DRX500
PROBHD
5 mm TBI 1H/
PULPROG
mlevetgp
TD
1024
SOLVENT
CDCl3
NS
8
DS
16
SWH
2185.315 Hz
FIDRES
2.134096 Hz
AQ
0.2345700 sec
RG
16
DW
228.800 usec
DE
6.00 usec
TE
300.0 K
D0
0.00000300 sec
D1
1.79999995 sec
D9
0.01000000 sec
D11
0.03000000 sec
D16
0.00010000 sec
DELTA
0.00109943 sec
DELTA1
0.00110800 sec
FACTOR1
0
IN0
0.00045765 sec
l1
0
l3
128
SCALEF
6

ppm

F1 - Processing parameters
SI
1024
MC2
echo-antiecho
SF
500.1300144 MHz
WDW
SINE
SSB
3
LB
0.00 Hz
GB
0

2D Gradient TOCSY, mixing time=20ms

1

2 3

Current Data Parameters

NAME
buttoc20
EXPNO
1
PROCNO
1

4
ppm

2-4 3-4

2 3

1.0

1-3
1-2

2-3
1.5

2.0

H1 H2 H3 H4
2.5

HO C C C C H
H H H H

Same as RLCOSY

3.0

3.0

2.5

2.0

1.5

============ CHANNEL f1
NUC1
1H
P1
5.60
P2
11.20
P5
19.33
P6
29.00
P7
58.00
P17
2500.00
PL1
-4.00
PL10
10.00
SFO1
500.1310815

=============
usec
usec
usec
usec
usec
usec
dB
dB
MHz

============ GRADIENT CHANNEL ========

GPNAM1
sine.100
GPNAM2
sine.100
GPX1
0.00 %
GPX2
0.00 %
GPY1
0.00 %
GPY2
0.00 %
GPZ1
30.00 %
GPZ2
30.00 %
P16
1000.00 usec
F1 - Acquisition parameters
ND0
1
TD
256
SFO1
500.1311 MHz
FIDRES
8.535453 Hz
SW
4.369 ppm
FnMODE
undefined
F2 - Processing parameters
SI
1024
SF
500.1300144 MHz
WDW
SINE
SSB
2
LB
0.00 Hz
GB
0
PC
0.20

3.5

3.5

F2 - Acquisition Parameters
Date_
20001102
Time
5.28
INSTRUM
DRX500
PROBHD
5 mm TBI 1H/
PULPROG
mlevetgp
TD
1024
SOLVENT
CDCl3
NS
8
DS
16
SWH
2185.315 Hz
FIDRES
2.134096 Hz
AQ
0.2345700 sec
RG
16
DW
228.800 usec
DE
6.00 usec
TE
300.0 K
D0
0.00000300 sec
D1
1.79999995 sec
D9
0.02000000 sec
D11
0.03000000 sec
D16
0.00010000 sec
DELTA
0.00109943 sec
DELTA1
0.00110800 sec
FACTOR1
1
IN0
0.00045765 sec
l1
6
l3
128
SCALEF
6

1.0

ppm

F1 - Processing parameters
SI
1024
MC2
echo-antiecho
SF
500.1300144 MHz
WDW
SINE
SSB
2
LB
0.00 Hz
GB
0

2D Gradient TOCSY, mixing time=50ms

1

2 3

Current Data Parameters

NAME
buttoc50
EXPNO
1
PROCNO
1

ppm

2-4 3-4

1-4

2 3

1.0

1-3
1-2

2-3
1.5

2.0

H1 H2 H3 H4

2.5

HO C C C C H
H H H H

3.0

3.5

3.0

2.5

2.0

1.5

============ CHANNEL f1
NUC1
1H
P1
5.60
P2
11.20
P5
19.33
P6
29.00
P7
58.00
P17
2500.00
PL1
-4.00
PL10
10.00
SFO1
500.1310815

=============
usec
usec
usec
usec
usec
usec
dB
dB
MHz

============ GRADIENT CHANNEL ========

GPNAM1
sine.100
GPNAM2
sine.100
GPX1
0.00 %
GPX2
0.00 %
GPY1
0.00 %
GPY2
0.00 %
GPZ1
30.00 %
GPZ2
30.00 %
P16
1000.00 usec
F1 - Acquisition parameters
ND0
1
TD
256
SFO1
500.1311 MHz
FIDRES
8.535453 Hz
SW
4.369 ppm
F2 - Processing parameters
SI
1024
SF
500.1300144 MHz
WDW
QSINE
SSB
4
LB
0.00 Hz
GB
0
PC
0.20

Total Correlation

3.5

F2 - Acquisition Parameters
Date_
20001101
Time
16.43
INSTRUM
DRX500
PROBHD
5 mm TBI 1H/
PULPROG
mlevetgp
TD
1024
SOLVENT
CDCl3
NS
8
DS
16
SWH
2185.315 Hz
FIDRES
2.134096 Hz
AQ
0.2343412 sec
RG
18
DW
228.800 usec
DE
6.00 usec
TE
300.0 K
D0
0.00000300 sec
D1
1.79999995 sec
D9
0.05000000 sec
D11
0.03000000 sec
D16
0.00010000 sec
DELTA
0.00109943 sec
DELTA1
0.00110800 sec
FACTOR1
4
IN0
0.00045765 sec
l1
24
l3
128
SCALEF
6

1.0

ppm

F1 - Processing parameters
SI
1024
MC2
echo-antiecho
SF
500.1300144 MHz
WDW
QSINE
SSB
4
LB
0.00 Hz
GB
0

DEPT: Distortionless Enhancement by Polarization Transfer

Heteronuclear expt.
Detection: 13C
Distinguish
CH, CH2, CH3
By suitable combination of
=45, 90 & 135 spectra
CH & CH3up
CH2 down

Only CH
All CHs

HETCOR (Heteronuclear chemical shift correlation, 1H - 13C COSY)

13C

AQ

1H

t1

t2

1H

decoupling
Removing JCH splittings

The standard pulse sequence for 13C-detected

1H-13C chemical shift correlation.
*But

Inverse experiment has the following Advantages:

increase sensitivity of detecting the less sensitive nuclei

1H is in the direct detection dimension => larger np =>
better resolution

HMQC (Heteronuclear Multiple Quantum Coherence)

HSQC (Heteronuclear Single Quantum Coherence)
t2

1H

AQ

13C

/2

/2

t1

/2

/2 GARP-1
13C

decoupling

Pulse sequence for 1H-detected 1H-13C Removing JCH splittings

correlation through single-quantum coherence

HSQC can give considerably better 13C resolution and sensitivity

than HMQC for CH2 groups of natural products
HMQC has simpler pulse sequence and it is more robust and easier
to perform.

HSQC spectrum: H-C correlated

1

2 3

H-dimension

Current Data Parameters

NAME
buthsqc
EXPNO
1
PROCNO
1

4
ppm

10

15
20
25

30

C-dimension

35
40

H1 H2 H3 H4

45

HO C C C C H

50

H H H H

55

60
65
3.5

3.0

2.5

2.0

1.5

(HMQC and HETCOR spectra look similar)

1.0

ppm

F2 - Acquisition Parameters
Date_
20001102
Time
10.31
INSTRUM
DRX500
PROBHD
5 mm TBI 1H/
PULPROG
invietgpsi
TD
1024
SOLVENT
CDCl3
NS
1
DS
16
SWH
2185.315 Hz
FIDRES
2.134096 Hz
AQ
0.2345700 sec
RG
2298.8
DW
228.800 usec
DE
6.00 usec
TE
300.0 K
D0
0.00000300 sec
D1
2.00000000 sec
D4
0.00170000 sec
D11
0.03000000 sec
D13
0.00000300 sec
D16
0.00010000 sec
D24
0.00090000 sec
DELTA
0.00116720 sec
DELTA1
0.00110700 sec
IN0
0.00003600 sec
l3
256
============ CHANNEL f1
NUC1
1H
P1
5.60
P2
11.20
P28
2500.00
PL1
-4.00
SFO1
500.1310815

=============

============ CHANNEL f2
CPDPRG2
garp
NUC2
13C
P3
17.70
P4
35.40
PCPD2
89.00
PL2
-1.00
PL12
13.00
SFO2
125.7633722

=============

usec
usec
usec
dB
MHz

usec
usec
usec
dB
dB
MHz

============ GRADIENT CHANNEL ========

GPNAM1
sine.100
GPNAM2
sine.100
GPX1
0.00 %
GPX2
0.00 %
GPY1
0.00 %
GPY2
0.00 %
GPZ1
80.00 %
GPZ2
20.10 %
P16
1000.00 usec
F1 - Acquisition parameters
ND0
2
TD
512
SFO1
125.7634 MHz
FIDRES
27.126736 Hz
SW
110.437 ppm
FnMODE
undefined
F2 - Processing parameters
SI
1024
SF
500.1300144 MHz
WDW
SINE
SSB
2
LB
0.00 Hz
GB
0
PC
0.20
F1 - Processing parameters
SI
1024
MC2
echo-antiecho
SF
125.7577969 MHz
WDW
SINE
SSB
2
LB
0.00 Hz
GB
0

HSQC-TOCSY

mixing time
SLy

1H

13C

SLy

MLEV17

/2

/2

t1

/2

/2

AQ

GARP-1

Pulse sequence for 1H-detected HSQC-TOCSY experiment

through single-quantum coherence.
Longer mixing time gives longer extend of spin-spin
correlations.

Gradient HSQC-TOCSY(mixing time=10ms)

1

H-dimension

2 3

Current Data Parameters

NAME
buthsto10
EXPNO
1
PROCNO
1

4
ppm

10

15
20
25

30

C-dimension

1-2

35
40

H1 H2 H3 H4

45

HO C C C C H

50
55

Same as HSQCCOSY spectrum

3.5

2.5

2.0

1.5

60
65

2-1
3.0

============ CHANNEL f1
NUC1
1H
P1
5.60
P2
11.20
P5
19.34
P6
29.00
P7
58.00
P17
2500.00
P28
2000.00
PL1
-4.00
PL10
10.00
SFO1
500.1310815
============ CHANNEL f2
CPDPRG2
garp
NUC2
13C
P3
15.70
P4
31.40
PCPD2
89.00
PL2
-2.00
PL12
13.00
SFO2
125.7633722

=============
usec
usec
usec
usec
usec
usec
usec
dB
dB
MHz
=============
usec
usec
usec
dB
dB
MHz

============ GRADIENT CHANNEL ========

P16
1000.00 usec

H H H H

1-1

F2 - Acquisition Parameters
Date_
20001101
Time
18.31
INSTRUM
DRX500
PROBHD
5 mm TBI 1H/
PULPROG
invietgsml
TD
1024
SOLVENT
CDCl3
NS
4
DS
16
SWH
2185.315 Hz
FIDRES
2.134096 Hz
AQ
0.2343412 sec
RG
2048
DW
228.800 usec
DE
6.00 usec
TE
300.0 K
CNST0
2.0000000
D0
0.00000300 sec
D1
1.79999995 sec
D4
0.00170000 sec
D11
0.03000000 sec
D12
0.00002000 sec
D13
0.00000300 sec
D16
0.00010000 sec
d20
0.00111720 sec
d21
0.00110700 sec
IN0
0.00003600 sec
L1
3
l3
128

1.0

ppm

F1 - Acquisition parameters
ND0
2
TD
256
SFO1
125.7634 MHz
FIDRES
54.253471 Hz
SW
110.437 ppm
F2 - Processing parameters
SI
1024
SF
500.1300144 MHz
WDW
SINE
SSB
2
LB
0.00 Hz
GB
0
PC
0.20
F1 - Processing parameters
SI
512
MC2
echo-antiecho
SF
125.7577961 MHz
WDW
SINE
SSB
2
LB
0.00 Hz
GB
0

Gradient HSQC-TOCSY(mixing time=20ms)

1

H-dimension

2 3

Current Data Parameters

NAME
buthsto20
EXPNO
1
PROCNO
1

4
ppm

10

15

1-3

20
25

30

C-dimension

1-2

35

H1 H2 H3 H4

40

HO C C C C H

45

F2 - Acquisition Parameters
Date_
20001101
Time
19.08
INSTRUM
DRX500
PROBHD
5 mm TBI 1H/
PULPROG
invietgsml
TD
1024
SOLVENT
CDCl3
NS
4
DS
16
SWH
2185.315 Hz
FIDRES
2.134096 Hz
AQ
0.2343412 sec
RG
2048
DW
228.800 usec
DE
6.00 usec
TE
300.0 K
CNST0
2.0000000
D0
0.00000300 sec
D1
1.79999995 sec
D4
0.00170000 sec
D11
0.03000000 sec
D12
0.00002000 sec
D13
0.00000300 sec
D16
0.00010000 sec
d20
0.00111720 sec
d21
0.00110700 sec
IN0
0.00003600 sec
L1
8
l3
128
============ CHANNEL f1
NUC1
1H
P1
5.60
P2
11.20
P5
19.34
P6
29.00
P7
58.00
P17
2500.00
P28
2000.00
PL1
-4.00
PL10
10.00
SFO1
500.1310815
============ CHANNEL f2
CPDPRG2
garp
NUC2
13C
P3
15.70
P4
31.40
PCPD2
89.00
PL2
-2.00
PL12
13.00
SFO2
125.7633722

=============
usec
usec
usec
usec
usec
usec
usec
dB
dB
MHz
=============
usec
usec
usec
dB
dB
MHz

============ GRADIENT CHANNEL ========

P16
1000.00 usec

H H H H

50
55

60

1-1
3.5

65

2-1 3-1
3.0

2.5

2.0

1.5

1.0

ppm

F1 - Acquisition parameters
ND0
2
TD
256
SFO1
125.7634 MHz
FIDRES
54.253471 Hz
SW
110.437 ppm
F2 - Processing parameters
SI
1024
SF
500.1300144 MHz
WDW
SINE
SSB
2
LB
0.00 Hz
GB
0
PC
0.20
F1 - Processing parameters
SI
512
MC2
echo-antiecho
SF
125.7577961 MHz
WDW
SINE
SSB
2
LB
0.00 Hz
GB
0

Gradient HSQC-TOCSY(mixing time=80ms)

1

H-dimension

2 3

Current Data Parameters

NAME
buthsto80
EXPNO
1
PROCNO
1

4
ppm

10

1-4

15

1-3

20
25

30

1-2

35

C-dimension

H1 H2 H 3 H 4

40

HO C C C C H

45
50
55

60

3.5

4-1

2-1 3-1
3.0

2.5

2.0

============ CHANNEL f1
NUC1
1H
P1
5.60
P2
11.20
P5
19.34
P6
29.00
P7
58.00
P17
2500.00
P28
2000.00
PL1
-4.00
PL10
10.00
SFO1
500.1310815
============ CHANNEL f2
CPDPRG2
garp
NUC2
13C
P3
15.70
P4
31.40
PCPD2
89.00
PL2
-2.00
PL12
13.00
SFO2
125.7633722

=============
usec
usec
usec
usec
usec
usec
usec
dB
dB
MHz
=============
usec
usec
usec
dB
dB
MHz

============ GRADIENT CHANNEL ========

P16
1000.00 usec

H H H H

1-1

F2 - Acquisition Parameters
Date_
20001101
Time
19.45
INSTRUM
DRX500
PROBHD
5 mm TBI 1H/
PULPROG
invietgsml
TD
1024
SOLVENT
CDCl3
NS
4
DS
16
SWH
2185.315 Hz
FIDRES
2.134096 Hz
AQ
0.2343412 sec
RG
2048
DW
228.800 usec
DE
6.00 usec
TE
300.0 K
CNST0
2.0000000
D0
0.00000300 sec
D1
1.79999995 sec
D4
0.00170000 sec
D11
0.03000000 sec
D12
0.00002000 sec
D13
0.00000300 sec
D16
0.00010000 sec
d20
0.00111720 sec
d21
0.00110700 sec
IN0
0.00003600 sec
L1
40
l3
128

1.5

1.0

65
ppm

F1 - Acquisition parameters
ND0
2
TD
256
SFO1
125.7634 MHz
FIDRES
54.253471 Hz
SW
110.437 ppm
F2 - Processing parameters
SI
1024
SF
500.1300133 MHz
WDW
SINE
SSB
2
LB
0.00 Hz
GB
0
PC
0.20
F1 - Processing parameters
SI
512
MC2
echo-antiecho
SF
125.7577961 MHz
WDW
SINE
SSB
2
LB
0.00 Hz
GB
0

HMBC (Heteronuclear Multiple-Bond Correlation Spectroscopy)

t1

1H

AQ

13C

Pulse sequence for HMBC

H1
C1

H1

C2

C3

C4

C1

C2, C3 and C4: Quaternary or protonated carbons

C2

C3

X: O, N

Long range connections or connections between spin systems

Gradient HMBC
3
5

2
1

H-dimension

C-dimension 3 2 1

Current Data Parameters

NAME
cfyhmbc
EXPNO
1
PROCNO
1

ppm
30
40
50
60
70

1 H3C

CH3
4

90

3-4
5

80

110

120

3-5

CH3

876 5

100

130

2-6
2-7

8.0

7.5

7.0

6.5

6.0

5.5

5.0

4.5

4.0

140

1-7
1-8

3.5

150
160
ppm

F2 - Acquisition Parameters
Date_
20001103
Time
7.36
INSTRUM
DRX500
PROBHD
5 mm TBI 1H/
PULPROG
inv4gplrnd
TD
1024
SOLVENT
CDCl3
NS
4
DS
16
SWH
3443.526 Hz
FIDRES
3.362818 Hz
AQ
0.1488800 sec
RG
11585.2
DW
145.200 usec
DE
6.00 usec
TE
300.0 K
D0
0.00000300 sec
D1
1.50000000 sec
D6
0.07000000 sec
D13
0.00000300 sec
D16
0.00050000 sec
IN0
0.00002380 sec
============ CHANNEL f1
NUC1
1H
P1
6.00
P2
12.00
PL1
-4.00
SFO1
500.1326379

=============

============ CHANNEL f2
NUC2
13C
P3
15.70
PL2
-2.00
SFO2
125.7693867

=============

usec
usec
dB
MHz

usec
dB
MHz

============ GRADIENT CHANNEL ========

GPNAM1
sine.100
GPNAM2
sine.100
GPNAM3
sine.100
GPX1
0.00 %
GPX2
0.00 %
GPX3
0.00 %
GPY1
0.00 %
GPY2
0.00 %
GPY3
0.00 %
GPZ1
50.00 %
GPZ2
30.00 %
GPZ3
40.10 %
P16
1000.00 usec
F1 - Acquisition parameters
ND0
2
TD
256
SFO1
125.7694 MHz
FIDRES
82.064079 Hz
SW
167.039 ppm
FnMODE
undefined
F2 - Processing parameters
SI
1024
SF
500.1300144 MHz
WDW
QSINE
SSB
0
LB
0.00 Hz
GB
0
PC
0.20
F1 - Processing parameters
SI
512
MC2
QF
SF
125.7577961 MHz
WDW
QSINE
SSB
0
LB
0.00 Hz
GB
0

2D NOESY (Nuclear Overhauser Enhancements SpectroscopY)

t1

t2

AQ

The NOESY pulse sequence.

C ~ C
Ha

Hb
r<0.5nm

VC*r -6, r<0.5nm

For resonance assignment, chemical structure elucidation &

3D structure determination

Gradient NOESY
Current Data Parameters
NAME
cfynoesy
EXPNO
1
PROCNO
1

ppm

3.5

3-CH3, 5-H

4.0

4.5

O
1 H 3C

CH3
N

5.5

5.0

6.0

CH3

6.5

7.0

7.5

7.5

7.0

6.5

6.0

5.5

5.0

4.5

4.0

3.5

8.0
ppm

F2 - Acquisition Parameters
Date_
20001103
Time
8.53
INSTRUM
DRX500
PROBHD
5 mm TBI 1H/
PULPROG
noesygptp
TD
1024
SOLVENT
CDCl3
NS
8
DS
16
SWH
3443.526 Hz
FIDRES
3.362818 Hz
AQ
0.1488800 sec
RG
64
DW
145.200 usec
DE
6.00 usec
TE
288.0 K
D0
0.00000300 sec
D1
1.60000002 sec
D8
0.40000001 sec
D16
0.00010000 sec
d20
0.19890000 sec
IN0
0.00014520 sec
============ CHANNEL f1
NUC1
1H
P1
6.00
P2
12.00
PL1
-4.00
SFO1
500.1326379

=============
usec
usec
dB
MHz

============ GRADIENT CHANNEL ========

GPNAM1
sine.100
GPNAM2
sine.100
GPX1
0.00 %
GPX2
0.00 %
GPY1
0.00 %
GPY2
0.00 %
GPZ1
40.00 %
GPZ2
-40.00 %
P16
1000.00 usec
F1 - Acquisition parameters
ND0
2
TD
256
SFO1
500.1326 MHz
FIDRES
13.451274 Hz
SW
6.885 ppm
FnMODE
undefined
F2 - Processing parameters
SI
1024
SF
500.1300144 MHz
WDW
SINE
SSB
2
LB
0.00 Hz
GB
0
PC
0.20
F1 - Processing parameters
SI
1024
MC2
TPPI
SF
500.1300144 MHz
WDW
SINE
SSB
2
LB
0.00 Hz
GB
0

1
2

CH2O

CH2 O

3
CH2 O

H2N C

C N C

C N C

H H

H H

OH

NOESY
gives sequential
assignment of peptides

2D ROESY pulse program

90

mixing time

t1

Spin-lock

t2

For small molecule NOE can be very small or zero

ROESY can be used in place of NOESY experiment. ROE
intensity is also related to the H-H distances.

Structure Determination Procedures

1D 1H & 13C & DEPT (+MSIRbasic chemical structure or
functional groups information
Establish 13C-1H connections by thru bond JCH couplings
HMQCHSQCHSQC-TOCSY experiments
Establish 1H-1H connection (spin systems or partial pieces
Decoupled 1H, 1D TOCSY, 2D 1H-1H COSY, TOCSY expts.
usually starts with well-resolved 1H signals
Long range connectionsconnecting spin systems & assigning
quaternary carbon
1D NOESY & 2D HMBC, NOESY, ROESY experiments
3D structure or conformation determination
1D NOESY & 2D NOESY, ROESY, (HSQC)-NOESY expts.