Structure Elucidation with Nuclear Magnetic Resonance (NMR)
===
Magnetic Resonance (NMR) Magnetic Resonance (NMR)
This was prepared from any material downloaded from internet.
NMR Applications in Chemistry
Chem. Shift J Coupling NMR
Structure spectrum
Peak Int.
NOE
CH 3 5'
6' CH 2 OH
Correlation 2
OO
1 6 1' H-H，C-H 1' OH 9
4'
4 OH
3' 2'
6'' O
5''
OH
5 OH
4''
1'' COOCH 8 7'' 2
3''
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.
Some common NMR experiments: Some common NMR experiments:
Connections through bonds Connections through bonds （ （ spin spin - - spin coupling spin coupling ） ）
1 1 1 Homonuclear 1 Homonuclear ： 1D ： 1D 、 、 2D 2D H H - - H COSY, DQF H COSY, DQF - - COSY, TOCSY COSY, TOCSY
usage usage ： ： spin system assignment spin system assignment
Heteronuclear Heteronuclear ： ：
13 Direct (detect 13 Direct (detect C): APT, DEPT, HETCOR C): APT, DEPT, HETCOR
1 Inverse (detect 1 Inverse (detect H): HMQC, HSQC, HMBC, HSQC H): HMQC, HSQC, HMBC, HSQC - - COSY, COSY, HSQC HSQC - - TOCSY, HMQC TOCSY, HMQC - - TOCSY TOCSY
usage usage ： ： assigning assigning heteronuclei heteronuclei 、 、 connecting spin systems connecting spin systems
Connections through space Connections through space （ （ dipolar coupling dipolar coupling ） ）
1D 1D 、 、 2D NOESY 2D NOESY ， ， ROESY, HOESY(HSQC ROESY, HOESY(HSQC - - NOESY) NOESY) usage usage ： ： connecting spin systems connecting spin systems 、 、 structure determination structure determination
2D COSY 2D COSY — — Homonuclear Homonuclear Shift CO Shift CO rrelation rrelation S S pectroscopY pectroscop Y
1 AQ
The basic COSY (x=45 ° or 90°) pulse sequence
Vicinal Geminal
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 2D Gradient COSY - - 45 45
Current Data Parameters
EXPNO 1 1 NAME butcosy 1 2 2 3 3 4 4
PROCNO 1 F2 - Acquisition Parameters
Date_ 20001102 Time 8.04
ppm
INSTRUM DRX500 PROBHD 5 mm TBI 1H/ PULPROG cosygp TD 1024 SOLVENT CDCl3
4 NS 1 4 3
3 - - 4 4 DS 16 SWH 2185.315 Hz FIDRES 2.134096 Hz AQ 0.2345700 sec
1.0 RG 40.3
DW 228.800 usec DE 6.00 usec TE 288.0 K
2 D0 0.00000300 sec 2 - - 3 3
3 D1 1.60000002 sec
3 D13 0.00000300 sec 1 1 - - 2 2
D16 0.00010000 sec
2 IN0 0.00045765 sec 2
1.5 ============ CHANNEL f1 ============= NUC1 1H P0 3.00 usec P1 6.00 usec PL1 -4.00 dB SFO1 500.1310815 MHz
============ GRADIENT CHANNEL ========
2.0 GPNAM1 sine.100 GPNAM2 sine.100 GPX1 0.00 %
H GPX2 0.00 %
GPY1 0.00 % GPY2 0.00 % GPZ1 10.00 % GPZ2 10.00 %
HO P16 1000.00 usec C C C C H
2.5 F1 - Acquisition parameters ND0 1
H TD 256 H H H
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 1 PC 0.20 1
3.5 F1 - Processing parameters SI 1024 SF 500.1300144 MHz MC2 QF
WDW QSINE SSB 0 LB 0.00 Hz
3.5 3.0 2.5 2.0 1.5 1.0 ppm
GB 0
2D 2D DQF DQF － － COSY COSY ( ( D D ouble ouble - - Q Q uantum uantum F F iltered iltered COSY COSY ) )
) ) allow J coupling constant measurements allow J coupling constant measurements ) ) observing cross observing cross - - peaks close to diagonal peaks close to diagonal ) ) But lower sensitivity (~40%) of COSY expt. But lower sensitivity (~40%) of COSY expt.
2D Gradient DQF 2D Gradient DQF - - COSY COSY
Current Data Parameters NAME gbc35dqf PROCNO 1 EXPNO 2
F2 - Acquisition Parameters Date_ 20001103 Time 4.01 INSTRUM DRX500
ppm
PROBHD 5 mm TBI 1H/ PULPROG cosydfetgp.2 TD 4096 NS 8 SOLVENT Pyr
3.6 SWH 3822.630 Hz DS 16 FIDRES 0.933259 Hz
AQ 0.5359376 sec
3.8 RG 512 DW 130.800 usec DE 6.00 usec
TE 288.0 K
4.0 D1 1.60000002 sec D0 0.00000300 sec D11 0.03000000 sec D16 0.00010000 sec
4.2 DELTA 0.00110300 sec
DELTA1 0.00110800 sec IN0 0.00026161 sec l3 256
4.4 ============ CHANNEL f1 ============= NUC1 1H P1 5.80 usec P2 11.60 usec PL1 -4.00 dB
4.6 SFO1 500.1317609 MHz ============ GRADIENT CHANNEL ========
GPNAM1 sine.100
4.8 GPNAM2 sine.100 GPNAM3 sine.100 GPX1 0.00 % GPX2 0.00 %
5.0 GPY1 0.00 % GPX3 0.00 % GPY2 0.00 % GPY3 0.00 %
5.2 GPZ1 30.00 %
GPZ2 10.00 % GPZ3 50.00 % P16 1000.00 usec
5.4 F1 - Acquisition parameters ND0 1 TD 512 SFO1 500.1318 MHz
SW 7.643 ppm 5.6 FIDRES 7.465717 Hz
FnMODE undefined
5.8 SI 4096 F2 - Processing parameters SF 500.1300038 MHz
SSB 2 WDW SINE
6.0 LB 0.00 Hz
GB 0 PC 0.20
6.2 F1 - Processing parameters
SI 1024 MC2 echo-antiecho SF 500.1300038 MHz
WDW SINE SSB 2 LB 0.00 Hz
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
GB 0
3 3 J=8.00Hz
J=8.00Hz
TOCSY TOCSY (TO ( TO tal C tal C orrelation S orrelation S pectroscopY pectroscop Y ) or ) or HOHAHA HOHAHA (Ho ( Ho monuclear Ha monuclear Ha rtman rtman -Ha - Ha hn Spectroscopy) hn Spectroscopy)
mixing time mixing time
MLEV17
AQ
Pulse sequence for a TOCSY spectrum.
H 1 H 2 H 3 H 4 H 1 H 2 H 3 H 4 H 1 H 2 H 3 H 4 HO C C C C H HO C C C C H HO C C C C H
COSY COSY
RL RL - - COSY COSY
TOCSY TOCSY
Different mixing time gives different degree of relay of Different mixing time gives different degree of relay of correlation. At small mixing time, TOCSY spectrum is similar to correlation. At small mixing time, TOCSY spectrum is similar to COSY spectrum. At long mixing time, gives total correlation. COSY spectrum. At long mixing time, gives total correlation.
2D Gradient TOCSY, 2D Gradient TOCSY, mixing time=10ms
Current Data Parameters NAME buttoc10 PROCNO 1 EXPNO 1
Date_ 20001102 1 F2 - Acquisition Parameters 1 2 2 3 3 4 4
Time 4.14 INSTRUM DRX500
ppm PROBHD 5 mm TBI 1H/
PULPROG mlevetgp TD 1024 SOLVENT CDCl3 NS 8 DS 16
SWH 2185.315 Hz FIDRES 2.134096 Hz
4 AQ 0.2345700 sec 4 3 RG 16 3 - - 4 4
DE 6.00 usec DW 228.800 usec
D9 0.01000000 sec D1 1.79999995 sec D0 0.00000300 sec 1.0 TE 300.0 K
D11 0.03000000 sec D16 0.00010000 sec DELTA 0.00109943 sec
2 DELTA1 0.00110800 sec 2 - - 3 3 FACTOR1 0 3 IN0 0.00045765 sec 3
1 l1 0 1 - - 2 2
1.5 SCALEF 6 2 l3 128 2
============ CHANNEL f1 ============= P1 5.60 usec NUC1 1H P2 11.20 usec P5 19.33 usec
P7 58.00 usec P6 29.00 usec P17 2500.00 usec
2.0 PL1 -4.00 dB
SFO1 500.1310815 MHz H PL10 10.00 dB
GPNAM1 sine.100 ============ GRADIENT CHANNEL ======== GPNAM2 sine.100 GPX1 0.00 %
HO GPX2 0.00 % C C C C H GPY1 0.00 %
2.5 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
3.0 FnMODE undefined
F2 - Processing parameters Same as COSY spectrum SI 1024 SSB 3 WDW SINE Same as COSY spectrum SF 500.1300144 MHz
GB 0 LB 0.00 Hz
1 PC 0.20 1 3.5
F1 - Processing parameters SI 1024 MC2 echo-antiecho SSB 3 WDW SINE SF 500.1300144 MHz LB 0.00 Hz
3.5 3.0 2.5 2.0 1.5 1.0 ppm
GB 0
2D Gradient TOCSY, 2D Gradient TOCSY, mixing time=20ms
Current Data Parameters EXPNO 1 NAME buttoc20
1 PROCNO 1 1 2 2 3 3 4 4
F2 - Acquisition Parameters Date_ 20001102 Time 5.28 PROBHD 5 mm TBI 1H/ INSTRUM DRX500
ppm
PULPROG mlevetgp TD 1024 SOLVENT CDCl3 NS 8 DS 16
FIDRES 2.134096 Hz 4 SWH 2185.315 Hz 4 2 2 - - 4 4 3 3 - - 4 4
AQ 0.2345700 sec RG 16 DW 228.800 usec
DE 6.00 usec
1.0 TE 300.0 K
D1 1.79999995 sec D0 0.00000300 sec D9 0.02000000 sec D11 0.03000000 sec DELTA 0.00109943 sec D16 0.00010000 sec DELTA1 0.00110800 sec
3 FACTOR1 1 3
IN0 0.00045765 sec l1 6
2 l3 128 2 1 1 - - 2 2 1.5 SCALEF 6
P2 11.20 usec P1 5.60 usec NUC1 1H ============ CHANNEL f1 ============= P6 29.00 usec P5 19.33 usec P17 2500.00 usec P7 58.00 usec
2.0 PL1 -4.00 dB SFO1 500.1310815 MHz PL10 10.00 dB
GPNAM1 sine.100 H ============ GRADIENT CHANNEL ======== GPNAM2 sine.100
GPX1 0.00 % GPX2 0.00 %
GPY2 0.00 % 2.5 GPY1 0.00 %
P16 1000.00 usec GPZ2 30.00 % HO GPZ1 30.00 % C C C C H
H TD 256 H H H
F1 - Acquisition parameters ND0 1
SFO1 500.1311 MHz FIDRES 8.535453 Hz
Same as RL Same as RL － － COSY COSY
3.0 SW 4.369 ppm FnMODE undefined
F2 - Processing parameters SI 1024
WDW SINE SF 500.1300144 MHz SSB 2
LB 0.00 Hz 1 GB 0 1
3.5 PC 0.20
F1 - Processing parameters SI 1024 MC2 echo-antiecho SF 500.1300144 MHz WDW SINE SSB 2 LB 0.00 Hz
3.5 3.0 2.5 2.0 1.5 1.0 ppm
GB 0
2D Gradient TOCSY, 2D Gradient TOCSY, mixing time=50ms
Current Data Parameters NAME buttoc50
PROCNO 1 1 EXPNO 1 1 2 2 3 3 4 4
F2 - Acquisition Parameters Date_ 20001101 INSTRUM DRX500 Time 16.43
PULPROG mlevetgp ppm PROBHD 5 mm TBI 1H/
TD 1024 SOLVENT CDCl3 NS 8 DS 16
FIDRES 2.134096 Hz 4 SWH 2185.315 Hz 4 2 2 - - 4 4 3 3 - - 4 4 AQ 0.2343412 sec
1 RG 18 1 - - 4 4 DW 228.800 usec
TE 300.0 K DE 6.00 usec
1.0 D0 0.00000300 sec D1 1.79999995 sec
D9 0.05000000 sec D11 0.03000000 sec
DELTA 0.00109943 sec 1 D16 0.00010000 sec 1 - - 3 3 2 2 - - 3 3 FACTOR1 4 3 DELTA1 0.00110800 sec 3 IN0 0.00045765 sec 2 l1 24
1.5 SCALEF 6 l3 128
============ CHANNEL f1 ============= NUC1 1H P2 11.20 usec P1 5.60 usec P7 58.00 usec P6 29.00 usec P5 19.33 usec
2.0 P17 2500.00 usec PL1 -4.00 dB PL10 10.00 dB SFO1 500.1310815 MHz
GPNAM1 sine.100 ============ GRADIENT CHANNEL ========
GPNAM2 sine.100 H GPX1 0.00 %
GPX2 0.00 % GPY1 0.00 %
2.5 GPY2 0.00 % GPZ1 30.00 %
P16 1000.00 usec HO GPZ2 30.00 % C C C C H
F1 - Acquisition parameters ND0 1
H TD 256 H H FIDRES 8.535453 Hz H SFO1 500.1311 MHz
3.0 SW 4.369 ppm F2 - Processing parameters
SI 1024 Total Correlation SF 500.1300144 MHz Total Correlation
WDW QSINE SSB 4
LB 0.00 Hz 1 GB 0 1
3.5 PC 0.20
F1 - Processing parameters SI 1024 MC2 echo-antiecho WDW QSINE SF 500.1300144 MHz SSB 4 LB 0.00 Hz
3.5 3.0 2.5 2.0 1.5 1.0 ppm
GB 0
DEPT : D istortionless E nhancement by P olarization T ransfer
Heteronuclear expt. Detection: 13 C
Distinguish CH, CH2, CH 3
By suitable combination of
θ=45, 90 & 135 spectra
CH & CH 3 up CH 2 down
Only CH All CH’s
1 1 13 HETCOR 13 HETCOR ( ( Het Het eronuclear eronuclear chemical shift chemical shift cor cor relation, relation, H H - - C COSY) C COSY)
13 C
AQ
H decoupling
Removing J CH splittings
13 The standard pulse sequence for 13 The standard pulse sequence for C C - - detected detected
1 1 13 H 13 H - - C chemical shift correlation. C chemical shift correlation.
* But Inverse experiment has the following Advantages: •increase sensitivity of detecting the less sensitive nuclei
• 1 H is in the direct detection dimension => larger np => better resolution
HMQC HMQC (H ( H eteronuclear M eteronuclear M ultiple Q ultiple Q uantum C uantum C oherence) oherence) HSQC HSQC (H ( H eteronuclear S eteronuclear S ingle ingle Q Q uantum C uantum C oherence) oherence)
AQ
13 C Δ/2
Δ/2 GARP-1
13 C decoupling
1 1 Pulse sequence for 13 H-detected H- C
Removing J CH splittings
correlation through single-quantum coherence
• HSQC can give considerably better 13 C resolution and sensitivity
than HMQC for CH 2 groups of natural products
• HMQC has simpler pulse sequence and it is more robust and easier to perform.
HSQC spectrum: H HSQC spectrum: H - - C correlated C correlated
EXPNO 1 NAME buthsqc Current Data Parameters
1 PROCNO 1 1 2 2 3 3 4 4 Date_ 20001102 F2 - Acquisition Parameters H Time 10.31 H - - dimension dimension
INSTRUM DRX500 PROBHD 5 mm TBI 1H/ ppm PULPROG invietgpsi TD 1024
SOLVENT CDCl3 NS 1
10 DS 16 SWH 2185.315 Hz
FIDRES 2.134096 Hz 4 AQ 0.2345700 sec 4
RG 2298.8 DW 228.800 usec DE 6.00 usec
15 TE 300.0 K D0 0.00000300 sec
D4 0.00170000 sec D1 2.00000000 sec
D16 0.00010000 sec D13 0.00000300 sec 20 D11 0.03000000 sec
DELTA 0.00116720 sec D24 0.00090000 sec IN0 0.00003600 sec DELTA1 0.00110700 sec
25 l3 256
============ CHANNEL f1 ============= NUC1 1H P2 11.20 usec P1 5.60 usec
SFO1 500.1310815 MHz PL1 -4.00 dB 30 P28 2500.00 usec
2 ============ CHANNEL f2 ============= 2 CPDPRG2 garp
P3 17.70 usec 35 NUC2 13C
PCPD2 89.00 usec P4 35.40 usec PL2 -1.00 dB SFO2 125.7633722 MHz PL12 13.00 dB
GPX1 0.00 % GPNAM2 sine.100 GPNAM1 sine.100 ============ GRADIENT CHANNEL ========
45 GPY1 0.00 % H GPX2 0.00 %
GPZ1 80.00 % GPY2 0.00 % P16 1000.00 usec GPZ2 20.10 %
50 HO F1 - Acquisition parameters C C C C H
TD 512 ND0 2 SFO1 125.7634 MHz
SW 110.437 ppm FIDRES 27.126736 Hz C-dimension FnMODE undefined
H H H H 55
SI 1024 F2 - Processing parameters
SSB 2 WDW SINE 60 SF 500.1300144 MHz
1 LB 0.00 Hz 1 GB 0
PC 0.20
65 F1 - Processing parameters
MC2 echo-antiecho SI 1024 WDW SINE SF 125.7577969 MHz LB 0.00 Hz SSB 2
3.5 3.0 2.5 2.0 1.5 1.0 ppm
GB 0
(HMQC and HETCOR spectra look similar) (HMQC and HETCOR spectra look similar)
HSQC HSQC - - TOCSY TOCSY
mixing time mixing time
SLy
SLy
MLEV17 AQ
13 C Δ/2
GARP-1
1 Pulse sequence for 1 Pulse sequence for H H - - detected HSQC detected HSQC - - TOCSY experiment TOCSY experiment through single through single - - quantum coherence. quantum coherence.
Longer mixing time gives longer extend of spin Longer mixing time gives longer extend of spin - - spin spin correlations. correlations.
Gradient HSQC Gradient HSQC - - TOCSY(mixing time=10ms) TOCSY(mixing time=10ms)
EXPNO 1 NAME buthsto10 Current Data Parameters
1 PROCNO 1 1 2 2 3 3 4 4
F2 - Acquisition Parameters Date_ 20001101
Time 18.31 H INSTRUM DRX500 H - - dimension dimension
PROBHD 5 mm TBI 1H/
ppm
PULPROG invietgsml TD 1024 SOLVENT CDCl3 NS 4
10 DS 16 SWH 2185.315 Hz FIDRES 2.134096 Hz
4 AQ 0.2343412 sec 4
DW 228.800 usec RG 2048
15 DE 6.00 usec TE 300.0 K
D0 0.00000300 sec 3 CNST0 2.0000000 3
D4 0.00170000 sec D1 1.79999995 sec
D16 0.00010000 sec D13 0.00000300 sec D12 0.00002000 sec D11 0.03000000 sec d20 0.00111720 sec d21 0.00110700 sec
25 L1 3 IN0 0.00003600 sec l3 128
============ CHANNEL f1 ============= P1 5.60 usec 30 NUC1 1H P2 11.20 usec
P7 58.00 usec P6 29.00 usec 2 P5 19.34 usec 2
35 P28 2000.00 usec 1 P17 2500.00 usec 1 - - 2 2
SFO1 500.1310815 MHz PL10 10.00 dB PL1 -4.00 dB
40 ============ CHANNEL f2 ============= CPDPRG2 garp
H NUC2 13C
3 P3 15.70 usec H
P4 31.40 usec PCPD2 89.00 usec
45 PL2 -2.00 dB PL12 13.00 dB
HO SFO2 125.7633722 MHz C C C C H
============ GRADIENT CHANNEL ======== P16 1000.00 usec
50 ND0 2 H F1 - Acquisition parameters H H H
TD 256 SW 110.437 ppm FIDRES 54.253471 Hz SFO1 125.7634 MHz
55 C-dimension F2 - Processing parameters
WDW SINE SF 500.1300144 MHz Same as HSQC SSB 2 Same as HSQC － － COSY
SI 1024
COSY spectrum spectrum
60 1 LB 0.00 Hz 1
PC 0.20 GB 0
65 MC2 echo-antiecho SI 512 F1 - Processing parameters
2 2 - - 1 1 WDW SINE SF 125.7577961 MHz
GB 0 LB 0.00 Hz 3.5 SSB 2 3.0 2.5 2.0 1.5 1.0 ppm
Gradient HSQC Gradient HSQC - - TOCSY(mixing time=20ms) TOCSY(mixing time=20ms)
PROCNO 1 EXPNO 1 NAME buthsto20 Current Data Parameters
2 2 3 3 4 4 F2 - Acquisition Parameters Time 19.08 Date_ 20001101 H INSTRUM DRX500 H - - dimension dimension
PROBHD 5 mm TBI 1H/
ppm
PULPROG invietgsml TD 1024 NS 4 SOLVENT CDCl3
10 DS 16 SWH 2185.315 Hz FIDRES 2.134096 Hz
4 AQ 0.2343412 sec 4 RG 2048
15 DW 228.800 usec DE 6.00 usec
D0 0.00000300 sec CNST0 2.0000000 TE 300.0 K
D1 1.79999995 sec 1 D4 0.00170000 sec 1 - - 3 3
20 D11 0.03000000 sec D16 0.00010000 sec D13 0.00000300 sec D12 0.00002000 sec d20 0.00111720 sec
25 IN0 0.00003600 sec d21 0.00110700 sec l3 128 L1 8
NUC1 1H ============ CHANNEL f1 =============
30 P1 5.60 usec P2 11.20 usec
P5 19.34 usec 2 P6 29.00 usec
P7 58.00 usec
1 P17 2500.00 usec 1 - - 2 2 35
PL1 -4.00 dB P28 2000.00 usec PL10 10.00 dB SFO1 500.1310815 MHz
40 ============ CHANNEL f2 ============= H CPDPRG2 garp
2 NUC2 13C 4 P3 15.70 usec P4 31.40 usec
PCPD2 89.00 usec PL2 -2.00 dB
45 HO PL12 13.00 dB C C C C H SFO2 125.7633722 MHz
============ GRADIENT CHANNEL ======== P16 1000.00 usec
50 H F1 - Acquisition parameters H H H ND0 2
TD 256 SFO1 125.7634 MHz FIDRES 54.253471 Hz
C-dimension
55 SW 110.437 ppm
F2 - Processing parameters
SI 1024 SF 500.1300144 MHz
60 SSB 2 WDW SINE LB 0.00 Hz GB 0
PC 0.20 F1 - Processing parameters
65 SI 512 1 MC2 echo-antiecho 1 - - 1 1 2 2 - - 1 1 3 3 - - 1 1
GB 0 LB 0.00 Hz SSB 2 3.5 WDW SINE 3.0 2.5 2.0 1.5 1.0 ppm
SF 125.7577961 MHz
Gradient HSQC Gradient HSQC - - TOCSY(mixing time=80ms) TOCSY(mixing time=80ms)
PROCNO 1 EXPNO 1 NAME buthsto80 Current Data Parameters
2 2 3 3 4 4 F2 - Acquisition Parameters Time 19.45 Date_ 20001101 H INSTRUM DRX500 H - - dimension dimension
PROBHD 5 mm TBI 1H/
ppm
PULPROG invietgsml TD 1024 NS 4 SOLVENT CDCl3
10 DS 16 SWH 2185.315 Hz FIDRES 2.134096 Hz
4 AQ 0.2343412 sec 4 RG 2048
15 DW 228.800 usec DE 6.00 usec TE 300.0 K D0 0.00000300 sec 3 CNST0 2.0000000 3 D1 1.79999995 sec
1 D4 0.00170000 sec 1 - - 3 3
20 D11 0.03000000 sec D12 0.00002000 sec D16 0.00010000 sec D13 0.00000300 sec d20 0.00111720 sec d21 0.00110700 sec
25 IN0 0.00003600 sec l3 128 L1 40
============ CHANNEL f1 ============= NUC1 1H
30 P1 5.60 usec P2 11.20 usec
P7 58.00 usec P6 29.00 usec P5 19.34 usec
1 P17 2500.00 usec 1 - - 2 2 35 P28 2000.00 usec
PL10 10.00 dB PL1 -4.00 dB SFO1 500.1310815 MHz
H 1 H 2 H 3 H 4 40
NUC2 13C CPDPRG2 garp ============ CHANNEL f2 ============= P3 15.70 usec P4 31.40 usec
HO PCPD2 89.00 usec C C C C H
PL12 13.00 dB 45 PL2 -2.00 dB
SFO2 125.7633722 MHz ============ GRADIENT CHANNEL ========
H P16 1000.00 usec H H H
50 F1 - Acquisition parameters ND0 2
TD 256 FIDRES 54.253471 Hz SFO1 125.7634 MHz SW 110.437 ppm
55 C-dimension F2 - Processing parameters
SI 1024 WDW SINE SF 500.1300133 MHz
60 GB 0 LB 0.00 Hz SSB 2
PC 0.20
65 SI 512 F1 - Processing parameters MC2 echo-antiecho
2 2 - - 1 1 3 3 - - 1 1 4 4 - - 1 1 WDW SINE SF 125.7577961 MHz
GB 0 LB 0.00 Hz 3.5 SSB 2 3.0 2.5 2.0 1.5 1.0 ppm
HMBC HMBC (H ( H eteronuclear M eteronuclear M ultiple ultiple -B - B ond C ond C orrelation Spectroscopy) orrelation Spectroscopy)
1 AQ
13 C
Pulse sequence for HMBC Pulse sequence for HMBC
C 2 ,C 3 and C 4 : Quaternary or protonated carbons X: O, N
Long range connections or connections between spin systems Long range connections or connections between spin systems
Gradient HMBC Gradient HMBC
2 2 Current Data Parameters NAME cfyhmbc
3 3 PROCNO 1 EXPNO 1
Date_ 20001103 F2 - Acquisition Parameters
INSTRUM DRX500 H Time 7.36 H - - dimension dimension
PULPROG inv4gplrnd ppm PROBHD 5 mm TBI 1H/
TD 1024 1 SOLVENT CDCl3 1
DS 16 NS 4
2 SWH 3443.526 Hz 2 30 FIDRES 3.362818 Hz AQ 0.1488800 sec
3 RG 11585.2 3 DW 145.200 usec
40 DE 6.00 usec D0 0.00000300 sec TE 300.0 K
D13 0.00000300 sec D6 0.07000000 sec 50 D1 1.50000000 sec
IN0 0.00002380 sec D16 0.00050000 sec
60 ============ CHANNEL f1 =============
NUC1 1H P1 6.00 usec P2 12.00 usec PL1 -4.00 dB
70 SFO1 500.1326379 MHz
P3 15.70 usec NUC2 13C 3 ============ CHANNEL f2 =============
80 PL2 -2.00 dB C-dimension SFO2 125.7693867 MHz O CH 3
GPNAM2 sine.100 GPNAM1 sine.100 90 ============ GRADIENT CHANNEL ========
GPNAM3 sine.100 1 GPX1 0.00 %
GPX3 0.00 % H GPX2 0.00 % 3 C
GPY2 0.00 % N GPY1 0.00 % 4 3 3 - - 4 4
GPY3 0.00 % 4 GPZ1 50.00 % 4
H GPZ3 40.10 %
GPZ2 30.00 %
7 P16 1000.00 usec
6 ND0 2 N TD 256
120 F1 - Acquisition parameters
SFO1 125.7694 MHz
FIDRES 82.064079 Hz SW 167.039 ppm
FnMODE undefined
3 F2 - Processing parameters 3 - - 5 5
5 SI 1024 140 5 SF 500.1300144 MHz CH
3 WDW QSINE 2 2 - - 6 6 SSB 0
6 LB 0.00 Hz 6 2 1 1 - - 7 7
PC 0.20 150 GB 0
7 7 F1 - Processing parameters 8 SI 512
2 2 - - 7 7 160 SF 125.7577961 MHz WDW QSINE
MC2 QF
1 1 - - 8 8 LB 0.00 Hz SSB 0
8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 ppm
GB 0
2D NOESY 2D NOESY (N ( N uclear O uclear O verhauser E verhauser E nhancements S nhancements S pectroscopY pectroscop Y ) ) AQ
The NOESY pulse sequence.
— C —~ — C —
∝C* r ,r <0.5nm
V -6
Ha Hb r <0.5nm
For resonance assignment, chemical structure elucidation & For resonance assignment, chemical structure elucidation &
3D structure determination 3D structure determination
Gradient NOESY Gradient NOESY
Current Data Parameters NAME cfynoesy EXPNO 1
PROCNO 1 F2 - Acquisition Parameters
Date_ 20001103 Time 8.53
ppm
INSTRUM DRX500 PROBHD 5 mm TBI 1H/ PULPROG noesygptp TD 1024 SOLVENT CDCl3 NS 8 DS 16 SWH 3443.526 Hz
3.5 FIDRES 3.362818 Hz AQ 0.1488800 sec RG 64
DW 145.200 usec TE 288.0 K DE 6.00 usec
3 3 - - CH3, 5 CH3, 5 - - H H
4.0 D0 0.00000300 sec D1 1.60000002 sec D8 0.40000001 sec
D16 0.00010000 sec IN0 0.00014520 sec d20 0.19890000 sec
4.5 ============ CHANNEL f1 =============
3 NUC1 1H
O P2 12.00 usec CH PL1 -4.00 dB
P1 6.00 usec
3 5.0 SFO1 500.1326379 MHz
1 ============ GRADIENT CHANNEL ========
H GPNAM1 sine.100
8 GPNAM2 sine.100 N N GPX1 0.00 % 4
5.5 GPX2 0.00 % GPY1 0.00 %
5 GPY2 0.00 % H
7 GPZ1 40.00 %
GPZ2 -40.00 %
6 P16 1000.00 usec N 6.0 O F1 - Acquisition parameters N
ND0 2 TD 256 SFO1 500.1326 MHz
CH SW 6.885 ppm
6.5 FIDRES 13.451274 Hz
FnMODE undefined
F2 - Processing parameters SI 1024
7.0 SF 500.1300144 MHz
WDW SINE SSB 2 LB 0.00 Hz
GB 0 PC 0.20
7.5 F1 - Processing parameters SI 1024 MC2 TPPI WDW SINE SF 500.1300144 MHz
8.0 SSB 2
LB 0.00 Hz
7.5 GB 0 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 ppm
CH 2 O
CH 2 O
CH 2 O
H N C C N C C N C C OH NOESY NOESY
2 gives sequential gives sequential
assignment of peptides assignment of peptides
2D ROESY pulse program 2D ROESY pulse program
mixing time mixing time
1 Spin-lock 2
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 Structure Determination Procedures
1 1 13 ¾ 13 ¾ 1D 1D H& H& C & DEPT (+MS C & DEPT (+MS 、 、 IR IR ， ， basic chemical structure or basic chemical structure or
functional groups information functional groups information ） ）
13 13 1 ¾ 1 ¾ Establish Establish C C - - H connections by thru bond J H connections by thru bond J CH CH couplings couplings HMQC HMQC 、 、 HSQC HSQC 、 、 HSQC HSQC - - TOCSY experiments TOCSY experiments
1 1 1 ¾ 1 ¾ Establish Establish H H - - H connection (spin systems or partial pieces H connection (spin systems or partial pieces ） ）
1 1 1 1 1 Decoupled 1 Decoupled H, 1D TOCSY, 2D H, 1D TOCSY, 2D H H - - H COSY, TOCSY H COSY, TOCSY expts expts . .
（usually starts with well （ usually starts with well - - resolved resolved H signals H signals ） ）
¾ ¾ Long range connections Long range connections （ （ connecting spin systems & assigning connecting spin systems & assigning
quaternary carbon quaternary carbon ） ）
1D NOESY & 2D HMBC, NOESY, ROESY experiments 1D NOESY & 2D HMBC, NOESY, ROESY experiments ¾ ¾ 3D structure or conformation determination 3D structure or conformation determination 1D NOESY & 2D NOESY, ROESY, (HSQC) 1D NOESY & 2D NOESY, ROESY, (HSQC) - - NOESY NOESY expts expts . .