nmr spectrum
DESCRIPTION
Chem. Shift. J Coupling. Peak Int. NOE. Correlation H-H , C-H. NMR Applications in Chemistry. NMR spectrum. Structure. Applications: • Sample quality control for Synthetic works . • Elucidation of chemical structures. - PowerPoint PPT PresentationTRANSCRIPT
NMR spectrum Structure
Chem. Shift
J Coupling
Peak Int.NOE
CorrelationH-H , C-H
O
OOH
OCH3
O
HOH
OH
OH
CH2OH
COOCH2
1
2
3
4
5
6
7
8
9
10
1''
2''3''
4''
5'' 6''
1'2'3'
4'
5'
6'
7''
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.
NMR Applications in Chemistry
Connections through spaceConnections through space (( dipolar couplingdipolar coupling )) 1D1D 、、 2D NOESY2D NOESY ,, ROESY, HOESY(HSQC-NOESY)ROESY, HOESY(HSQC-NOESY)
usageusage : : connecting spin systems connecting spin systems 、、 structure determinatistructure determinati
onon
Connections through bondsConnections through bonds (( spin-spin couplingspin-spin coupling )) HomonuclearHomonuclear : : 1D1D 、、 2D 2D 11H-H-11H COSY, DQF-COSY, TOCSYH COSY, DQF-COSY, TOCSY
usageusage :: spin system assignmentspin system assignment
HeteronuclearHeteronuclear : : Direct (detect Direct (detect 1313C): C): APT, DEPT, HETCORAPT, DEPT, HETCOR
Inverse (detectInverse (detect11H): H): HMQC, HSQC, HMBC, HSQC-COSY, HMQC, HSQC, HMBC, HSQC-COSY,
HSQC-TOCSY, HMQC-TOCSYHSQC-TOCSY, HMQC-TOCSY
usageusage :: assigning heteronucleiassigning heteronuclei 、、 connecting spin systemsconnecting spin systems
Some common NMR experiments:Some common NMR experiments:
COSY spectrum is used for determining the connectivitiesbetween protons on the basis of geminal and vicinal couplings.Disadvantage: bulky dispersive diagonal peaks.
C C H
HHVicinal Geminal
2D COSY2D COSY — Homonuclear Shift — Homonuclear Shift COCOrrelation rrelation SSpectroscoppectroscopYY
t1 AQ
The basic COSY (x=45° or 90°) pulse sequence
90° x° t2
ppm
1.01.52.02.53.03.5 ppm
1.0
1.5
2.0
2.5
3.0
3.5
Current Data ParametersNAME butcosyEXPNO 1PROCNO 1
F2 - Acquisition ParametersDate_ 20001102Time 8.04INSTRUM DRX500PROBHD 5 mm TBI 1H/PULPROG cosygpTD 1024SOLVENT CDCl3NS 1DS 16SWH 2185.315 HzFIDRES 2.134096 HzAQ 0.2345700 secRG 40.3DW 228.800 usecDE 6.00 usecTE 288.0 KD0 0.00000300 secD1 1.60000002 secD13 0.00000300 secD16 0.00010000 secIN0 0.00045765 sec
============ CHANNEL f1 =============NUC1 1HP0 3.00 usecP1 6.00 usecPL1 -4.00 dBSFO1 500.1310815 MHz
============ GRADIENT CHANNEL ========GPNAM1 sine.100GPNAM2 sine.100GPX1 0.00 %GPX2 0.00 %GPY1 0.00 %GPY2 0.00 %GPZ1 10.00 %GPZ2 10.00 %P16 1000.00 usec
F1 - Acquisition parametersND0 1TD 256SFO1 500.1311 MHzFIDRES 8.535453 HzSW 4.369 ppmFnMODE undefined
F2 - Processing parametersSI 2048SF 500.1300144 MHzWDW QSINESSB 0LB 0.00 HzGB 0PC 0.20
F1 - Processing parametersSI 1024MC2 QFSF 500.1300144 MHzWDW QSINESSB 0LB 0.00 HzGB 0
11 22 33 4411
2233
44
HO C
H1
H
C C C
H2
H
H3
H
H4
H
H
1-21-22-32-3
3-43-4
2D Gradient COSY-452D Gradient COSY-45
2D 2D DQFDQF -- COSYCOSY((DDouble-ouble-QQuantum uantum FFiltered iltered COSYCOSY))
90 90 90
t1 t2
allow J coupling constant measurements allow J coupling constant measurements observing cross-peaks close to diagonalobserving cross-peaks close to diagonal But lower sensitivity (~40%) of COSY expt.But lower sensitivity (~40%) of COSY expt.
ppm
3.63.84.04.24.44.64.85.05.25.45.65.86.06.2 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
Current Data ParametersNAME gbc35dqfEXPNO 2PROCNO 1
F2 - Acquisition ParametersDate_ 20001103Time 4.01INSTRUM DRX500PROBHD 5 mm TBI 1H/PULPROG cosydfetgp.2TD 4096SOLVENT PyrNS 8DS 16SWH 3822.630 HzFIDRES 0.933259 HzAQ 0.5359376 secRG 512DW 130.800 usecDE 6.00 usecTE 288.0 KD0 0.00000300 secD1 1.60000002 secD11 0.03000000 secD16 0.00010000 secDELTA 0.00110300 secDELTA1 0.00110800 secIN0 0.00026161 secl3 256
============ CHANNEL f1 =============NUC1 1HP1 5.80 usecP2 11.60 usecPL1 -4.00 dBSFO1 500.1317609 MHz
============ GRADIENT CHANNEL ========GPNAM1 sine.100GPNAM2 sine.100GPNAM3 sine.100GPX1 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 parametersND0 1TD 512SFO1 500.1318 MHzFIDRES 7.465717 HzSW 7.643 ppmFnMODE undefined
F2 - Processing parametersSI 4096SF 500.1300038 MHzWDW SINESSB 2LB 0.00 HzGB 0PC 0.20
F1 - Processing parametersSI 1024MC2 echo-antiechoSF 500.1300038 MHzWDW SINESSB 2LB 0.00 HzGB 0
2D Gradient DQF-COSY2D Gradient DQF-COSY
33J=8.00HzJ=8.00Hz
TOCSYTOCSY ( (TOTOtal tal CCorrelationorrelation SSpectroscoppectroscopYY) or) orHOHAHAHOHAHA((HoHomonuclear monuclear HaHartman-rtman-HaHahn Spectroscopy)hn Spectroscopy)
t1 MLEV17 AQ
Pulse sequence for a TOCSY spectrum.
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.
HO C
H1
H
C C C
H2
H
H3
H
H4
H
H
HO C
H1
H
C C C
H2
H
H3
H
H4
H
HHO C
H1
H
C C C
H2
H
H3
H
H4
H
H
COSYCOSY RL-COSYRL-COSY TOCSYTOCSY
mixing timemixing time t2
ppm
1.01.52.02.53.03.5 ppm
1.0
1.5
2.0
2.5
3.0
3.5
Current Data ParametersNAME buttoc10EXPNO 1PROCNO 1
F2 - Acquisition ParametersDate_ 20001102Time 4.14INSTRUM DRX500PROBHD 5 mm TBI 1H/PULPROG mlevetgpTD 1024SOLVENT CDCl3NS 8DS 16SWH 2185.315 HzFIDRES 2.134096 HzAQ 0.2345700 secRG 16DW 228.800 usecDE 6.00 usecTE 300.0 KD0 0.00000300 secD1 1.79999995 secD9 0.01000000 secD11 0.03000000 secD16 0.00010000 secDELTA 0.00109943 secDELTA1 0.00110800 secFACTOR1 0IN0 0.00045765 secl1 0l3 128SCALEF 6
============ CHANNEL f1 =============NUC1 1HP1 5.60 usecP2 11.20 usecP5 19.33 usecP6 29.00 usecP7 58.00 usecP17 2500.00 usecPL1 -4.00 dBPL10 10.00 dBSFO1 500.1310815 MHz
============ GRADIENT CHANNEL ========GPNAM1 sine.100GPNAM2 sine.100GPX1 0.00 %GPX2 0.00 %GPY1 0.00 %GPY2 0.00 %GPZ1 30.00 %GPZ2 30.00 %P16 1000.00 usec
F1 - Acquisition parametersND0 1TD 256SFO1 500.1311 MHzFIDRES 8.535453 HzSW 4.369 ppmFnMODE undefined
F2 - Processing parametersSI 1024SF 500.1300144 MHzWDW SINESSB 3LB 0.00 HzGB 0PC 0.20
F1 - Processing parametersSI 1024MC2 echo-antiechoSF 500.1300144 MHzWDW SINESSB 3LB 0.00 HzGB 0
HO C
H1
H
C C C
H2
H
H3
H
H4
H
H
2D Gradient TOCSY, 2D Gradient TOCSY, mixing time=10ms
Same as COSY spectrumSame as COSY spectrum
11 22 33 4411
2233
44
1-21-22-32-3
3-43-4
ppm
1.01.52.02.53.03.5 ppm
1.0
1.5
2.0
2.5
3.0
3.5
Current Data ParametersNAME buttoc20EXPNO 1PROCNO 1
F2 - Acquisition ParametersDate_ 20001102Time 5.28INSTRUM DRX500PROBHD 5 mm TBI 1H/PULPROG mlevetgpTD 1024SOLVENT CDCl3NS 8DS 16SWH 2185.315 HzFIDRES 2.134096 HzAQ 0.2345700 secRG 16DW 228.800 usecDE 6.00 usecTE 300.0 KD0 0.00000300 secD1 1.79999995 secD9 0.02000000 secD11 0.03000000 secD16 0.00010000 secDELTA 0.00109943 secDELTA1 0.00110800 secFACTOR1 1IN0 0.00045765 secl1 6l3 128SCALEF 6
============ CHANNEL f1 =============NUC1 1HP1 5.60 usecP2 11.20 usecP5 19.33 usecP6 29.00 usecP7 58.00 usecP17 2500.00 usecPL1 -4.00 dBPL10 10.00 dBSFO1 500.1310815 MHz
============ GRADIENT CHANNEL ========GPNAM1 sine.100GPNAM2 sine.100GPX1 0.00 %GPX2 0.00 %GPY1 0.00 %GPY2 0.00 %GPZ1 30.00 %GPZ2 30.00 %P16 1000.00 usec
F1 - Acquisition parametersND0 1TD 256SFO1 500.1311 MHzFIDRES 8.535453 HzSW 4.369 ppmFnMODE undefined
F2 - Processing parametersSI 1024SF 500.1300144 MHzWDW SINESSB 2LB 0.00 HzGB 0PC 0.20
F1 - Processing parametersSI 1024MC2 echo-antiechoSF 500.1300144 MHzWDW SINESSB 2LB 0.00 HzGB 0
2D Gradient TOCSY, 2D Gradient TOCSY, mixing time=20ms
11 22 33 4411
2233
44
1-21-22-32-3
3-43-4
1-31-3
2-42-4
Same as RLSame as RL -- COSYCOSY
HO C
H1
H
C C C
H2
H
H3
H
H4
H
H
ppm
1.01.52.02.53.03.5 ppm
1.0
1.5
2.0
2.5
3.0
3.5
Current Data ParametersNAME buttoc50EXPNO 1PROCNO 1
F2 - Acquisition ParametersDate_ 20001101Time 16.43INSTRUM DRX500PROBHD 5 mm TBI 1H/PULPROG mlevetgpTD 1024SOLVENT CDCl3NS 8DS 16SWH 2185.315 HzFIDRES 2.134096 HzAQ 0.2343412 secRG 18DW 228.800 usecDE 6.00 usecTE 300.0 KD0 0.00000300 secD1 1.79999995 secD9 0.05000000 secD11 0.03000000 secD16 0.00010000 secDELTA 0.00109943 secDELTA1 0.00110800 secFACTOR1 4IN0 0.00045765 secl1 24l3 128SCALEF 6
============ CHANNEL f1 =============NUC1 1HP1 5.60 usecP2 11.20 usecP5 19.33 usecP6 29.00 usecP7 58.00 usecP17 2500.00 usecPL1 -4.00 dBPL10 10.00 dBSFO1 500.1310815 MHz
============ GRADIENT CHANNEL ========GPNAM1 sine.100GPNAM2 sine.100GPX1 0.00 %GPX2 0.00 %GPY1 0.00 %GPY2 0.00 %GPZ1 30.00 %GPZ2 30.00 %P16 1000.00 usec
F1 - Acquisition parametersND0 1TD 256SFO1 500.1311 MHzFIDRES 8.535453 HzSW 4.369 ppm
F2 - Processing parametersSI 1024SF 500.1300144 MHzWDW QSINESSB 4LB 0.00 HzGB 0PC 0.20
F1 - Processing parametersSI 1024MC2 echo-antiechoSF 500.1300144 MHzWDW QSINESSB 4LB 0.00 HzGB 0
2D Gradient TOCSY, 2D Gradient TOCSY, mixing time=50ms
11 22 33 4411
2233
44
1-21-22-32-3
3-43-4
1-31-3
2-42-41-41-4
HO C
H1
H
C C C
H2
H
H3
H
H4
H
H
Total CorrelationTotal Correlation
DEPT: Distortionless Enhancement by Polarization Transfer Heteronuclear expt.
Detection: 13C
DistinguishCH, CH2, CH3
By suitable combination of =45, 90 & 135 spectra
All CH’s
Only CH
CH & CH3up CH2 down
HETCORHETCOR ( (HetHeteronuclear chemical shift eronuclear chemical shift corcorrelation, relation, 11H - H - 1313C COSY)C COSY)
13C
1H t1 1 2
The standard pulse sequence for The standard pulse sequence for 1313C-detectedC-detected11H-H-1313C chemical shift correlation.C chemical shift correlation.
AQt2
1H decouplingRemoving JCH splittings
*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
HSQCHSQC ( (HHeteronuclear eteronuclear SSingle ingle QQuantum uantum CCoherence)oherence)
AQ
1H
13C /2 /2 t1 /2 /2 GARP-1
Pulse sequence for 1H-detected 1H-13C 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.
13C decouplingRemoving JCH splittings
HMQCHMQC ( (HHeteronuclear eteronuclear MMultiple ultiple QQuantum uantum CCoherence)oherence)
t2
ppm
1.01.52.02.53.03.5 ppm
10
15
20
25
30
35
40
45
50
55
60
65
Current Data ParametersNAME buthsqcEXPNO 1PROCNO 1
F2 - Acquisition ParametersDate_ 20001102Time 10.31INSTRUM DRX500PROBHD 5 mm TBI 1H/PULPROG invietgpsiTD 1024SOLVENT CDCl3NS 1DS 16SWH 2185.315 HzFIDRES 2.134096 HzAQ 0.2345700 secRG 2298.8DW 228.800 usecDE 6.00 usecTE 300.0 KD0 0.00000300 secD1 2.00000000 secD4 0.00170000 secD11 0.03000000 secD13 0.00000300 secD16 0.00010000 secD24 0.00090000 secDELTA 0.00116720 secDELTA1 0.00110700 secIN0 0.00003600 secl3 256
============ CHANNEL f1 =============NUC1 1HP1 5.60 usecP2 11.20 usecP28 2500.00 usecPL1 -4.00 dBSFO1 500.1310815 MHz
============ CHANNEL f2 =============CPDPRG2 garpNUC2 13CP3 17.70 usecP4 35.40 usecPCPD2 89.00 usecPL2 -1.00 dBPL12 13.00 dBSFO2 125.7633722 MHz
============ GRADIENT CHANNEL ========GPNAM1 sine.100GPNAM2 sine.100GPX1 0.00 %GPX2 0.00 %GPY1 0.00 %GPY2 0.00 %GPZ1 80.00 %GPZ2 20.10 %P16 1000.00 usec
F1 - Acquisition parametersND0 2TD 512SFO1 125.7634 MHzFIDRES 27.126736 HzSW 110.437 ppmFnMODE undefined
F2 - Processing parametersSI 1024SF 500.1300144 MHzWDW SINESSB 2LB 0.00 HzGB 0PC 0.20
F1 - Processing parametersSI 1024MC2 echo-antiechoSF 125.7577969 MHzWDW SINESSB 2LB 0.00 HzGB 0
HO C
H1
H
C C C
H2
H
H3
H
H4
H
H
11 22 33 4411
2233
44HSQC spectrum: H-C correlatedHSQC spectrum: H-C correlated
(HMQC and HETCOR spectra look similar)(HMQC and HETCOR spectra look similar)
C-dime
nsion
H-dimensionH-dimension
HSQC-TOCSYHSQC-TOCSY
AQ1H
13C /2 /2 t1 /2 /2 GARP-1
Pulse sequence for Pulse sequence for 11H-detected HSQC-TOCSY experimentH-detected HSQC-TOCSY experimentthrough single-quantum coherence.through single-quantum coherence.
Longer mixing time gives longer extend of spin-spin Longer mixing time gives longer extend of spin-spin correlations.correlations.
SLy
MLEV17
SLy
mixing timemixing time
ppm
1.01.52.02.53.03.5 ppm
10
15
20
25
30
35
40
45
50
55
60
65
Current Data ParametersNAME buthsto10EXPNO 1PROCNO 1
F2 - Acquisition ParametersDate_ 20001101Time 18.31INSTRUM DRX500PROBHD 5 mm TBI 1H/PULPROG invietgsmlTD 1024SOLVENT CDCl3NS 4DS 16SWH 2185.315 HzFIDRES 2.134096 HzAQ 0.2343412 secRG 2048DW 228.800 usecDE 6.00 usecTE 300.0 KCNST0 2.0000000D0 0.00000300 secD1 1.79999995 secD4 0.00170000 secD11 0.03000000 secD12 0.00002000 secD13 0.00000300 secD16 0.00010000 secd20 0.00111720 secd21 0.00110700 secIN0 0.00003600 secL1 3l3 128
============ CHANNEL f1 =============NUC1 1HP1 5.60 usecP2 11.20 usecP5 19.34 usecP6 29.00 usecP7 58.00 usecP17 2500.00 usecP28 2000.00 usecPL1 -4.00 dBPL10 10.00 dBSFO1 500.1310815 MHz
============ CHANNEL f2 =============CPDPRG2 garpNUC2 13CP3 15.70 usecP4 31.40 usecPCPD2 89.00 usecPL2 -2.00 dBPL12 13.00 dBSFO2 125.7633722 MHz
============ GRADIENT CHANNEL ========P16 1000.00 usec
F1 - Acquisition parametersND0 2TD 256SFO1 125.7634 MHzFIDRES 54.253471 HzSW 110.437 ppm
F2 - Processing parametersSI 1024SF 500.1300144 MHzWDW SINESSB 2LB 0.00 HzGB 0PC 0.20
F1 - Processing parametersSI 512MC2 echo-antiechoSF 125.7577961 MHzWDW SINESSB 2LB 0.00 HzGB 0
HO C
H1
H
C C C
H2
H
H3
H
H4
H
H
Same as Same as HSQCHSQC -- COSYCOSY spectrum spectrum
Gradient HSQC-TOCSY(mixing time=10ms)Gradient HSQC-TOCSY(mixing time=10ms)11 22 33 44
1122
3344
1-11-1
1-21-2
2-12-1
H-dimensionH-dimensionC-d
imensi
on
ppm
1.01.52.02.53.03.5 ppm
10
15
20
25
30
35
40
45
50
55
60
65
Current Data ParametersNAME buthsto20EXPNO 1PROCNO 1
F2 - Acquisition ParametersDate_ 20001101Time 19.08INSTRUM DRX500PROBHD 5 mm TBI 1H/PULPROG invietgsmlTD 1024SOLVENT CDCl3NS 4DS 16SWH 2185.315 HzFIDRES 2.134096 HzAQ 0.2343412 secRG 2048DW 228.800 usecDE 6.00 usecTE 300.0 KCNST0 2.0000000D0 0.00000300 secD1 1.79999995 secD4 0.00170000 secD11 0.03000000 secD12 0.00002000 secD13 0.00000300 secD16 0.00010000 secd20 0.00111720 secd21 0.00110700 secIN0 0.00003600 secL1 8l3 128
============ CHANNEL f1 =============NUC1 1HP1 5.60 usecP2 11.20 usecP5 19.34 usecP6 29.00 usecP7 58.00 usecP17 2500.00 usecP28 2000.00 usecPL1 -4.00 dBPL10 10.00 dBSFO1 500.1310815 MHz
============ CHANNEL f2 =============CPDPRG2 garpNUC2 13CP3 15.70 usecP4 31.40 usecPCPD2 89.00 usecPL2 -2.00 dBPL12 13.00 dBSFO2 125.7633722 MHz
============ GRADIENT CHANNEL ========P16 1000.00 usec
F1 - Acquisition parametersND0 2TD 256SFO1 125.7634 MHzFIDRES 54.253471 HzSW 110.437 ppm
F2 - Processing parametersSI 1024SF 500.1300144 MHzWDW SINESSB 2LB 0.00 HzGB 0PC 0.20
F1 - Processing parametersSI 512MC2 echo-antiechoSF 125.7577961 MHzWDW SINESSB 2LB 0.00 HzGB 0
11 22 33 4411
2233
44
HO C
H1
H
C C C
H2
H
H3
H
H4
H
H
Gradient HSQC-TOCSY(mixing time=20ms)Gradient HSQC-TOCSY(mixing time=20ms)
1-21-2
2-12-11-11-1
1-31-3
3-13-1
C-dime
nsion
H-dimensionH-dimension
ppm
1.01.52.02.53.03.5 ppm
10
15
20
25
30
35
40
45
50
55
60
65
Current Data ParametersNAME buthsto80EXPNO 1PROCNO 1
F2 - Acquisition ParametersDate_ 20001101Time 19.45INSTRUM DRX500PROBHD 5 mm TBI 1H/PULPROG invietgsmlTD 1024SOLVENT CDCl3NS 4DS 16SWH 2185.315 HzFIDRES 2.134096 HzAQ 0.2343412 secRG 2048DW 228.800 usecDE 6.00 usecTE 300.0 KCNST0 2.0000000D0 0.00000300 secD1 1.79999995 secD4 0.00170000 secD11 0.03000000 secD12 0.00002000 secD13 0.00000300 secD16 0.00010000 secd20 0.00111720 secd21 0.00110700 secIN0 0.00003600 secL1 40l3 128
============ CHANNEL f1 =============NUC1 1HP1 5.60 usecP2 11.20 usecP5 19.34 usecP6 29.00 usecP7 58.00 usecP17 2500.00 usecP28 2000.00 usecPL1 -4.00 dBPL10 10.00 dBSFO1 500.1310815 MHz
============ CHANNEL f2 =============CPDPRG2 garpNUC2 13CP3 15.70 usecP4 31.40 usecPCPD2 89.00 usecPL2 -2.00 dBPL12 13.00 dBSFO2 125.7633722 MHz
============ GRADIENT CHANNEL ========P16 1000.00 usec
F1 - Acquisition parametersND0 2TD 256SFO1 125.7634 MHzFIDRES 54.253471 HzSW 110.437 ppm
F2 - Processing parametersSI 1024SF 500.1300133 MHzWDW SINESSB 2LB 0.00 HzGB 0PC 0.20
F1 - Processing parametersSI 512MC2 echo-antiechoSF 125.7577961 MHzWDW SINESSB 2LB 0.00 HzGB 0
HO C
H1
H
C C C
H2
H
H3
H
H4
H
H
11 22 33 4411
2233
44Gradient HSQC-TOCSY(mixing time=80ms)Gradient HSQC-TOCSY(mixing time=80ms)
1-21-2
2-12-11-11-1
1-31-3
3-13-1
1-41-4
4-14-1
C-dime
nsion
H-dimensionH-dimension
HMBCHMBC ( (HHeteronuclear eteronuclear MMultiple-ultiple-BBond ond CCorrelation Spectroscopy)orrelation Spectroscopy)
13C
1H t1 AQ
C2, C3 and C4: Quaternary or protonated carbons X: O, N
C1 C2 C3 C4
H1
C1 X C2 C3
H1
Pulse sequence for HMBCPulse sequence for HMBC
Long range connections or connections between spin systemsLong range connections or connections between spin systems
ppm
3.54.04.55.05.56.06.57.07.58.0 ppm
30
40
50
60
70
80
90
100
110
120
130
140
150
160
Current Data ParametersNAME cfyhmbcEXPNO 1PROCNO 1
F2 - Acquisition ParametersDate_ 20001103Time 7.36INSTRUM DRX500PROBHD 5 mm TBI 1H/PULPROG inv4gplrndTD 1024SOLVENT CDCl3NS 4DS 16SWH 3443.526 HzFIDRES 3.362818 HzAQ 0.1488800 secRG 11585.2DW 145.200 usecDE 6.00 usecTE 300.0 KD0 0.00000300 secD1 1.50000000 secD6 0.07000000 secD13 0.00000300 secD16 0.00050000 secIN0 0.00002380 sec
============ CHANNEL f1 =============NUC1 1HP1 6.00 usecP2 12.00 usecPL1 -4.00 dBSFO1 500.1326379 MHz
============ CHANNEL f2 =============NUC2 13CP3 15.70 usecPL2 -2.00 dBSFO2 125.7693867 MHz
============ GRADIENT CHANNEL ========GPNAM1 sine.100GPNAM2 sine.100GPNAM3 sine.100GPX1 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 parametersND0 2TD 256SFO1 125.7694 MHzFIDRES 82.064079 HzSW 167.039 ppmFnMODE undefined
F2 - Processing parametersSI 1024SF 500.1300144 MHzWDW QSINESSB 0LB 0.00 HzGB 0PC 0.20
F1 - Processing parametersSI 512MC2 QFSF 125.7577961 MHzWDW QSINESSB 0LB 0.00 HzGB 0
N
N N
N
O
O
CH3
CH3
H3C
H
1
3
5
2
8
76
4
Gradient HMBCGradient HMBC11
223355
1144
3322
5566
7788
3-43-4
3-53-5 2-62-62-72-7
1-71-71-81-8
H-dimensionH-dimensionC-d
imensi
on
2D 2D NOESY NOESY ((NNuclear uclear OOverhauser verhauser EEnhancements nhancements SSpectroscoppectroscopYY))
t1 m AQ
The NOESY pulse sequence.
—C — ~ —C —
Ha HbVC*r -6, r0.5nm
r0.5nm
For resonance assignment, chemical structure elucidation & For resonance assignment, chemical structure elucidation & 3D structure determination3D structure determination
t2
ppm
3.54.04.55.05.56.06.57.07.5 ppm
3.5
4.0
4.5
5.0
5.5
6.0
6.5
7.0
7.5
8.0
Current Data ParametersNAME cfynoesyEXPNO 1PROCNO 1
F2 - Acquisition ParametersDate_ 20001103Time 8.53INSTRUM DRX500PROBHD 5 mm TBI 1H/PULPROG noesygptpTD 1024SOLVENT CDCl3NS 8DS 16SWH 3443.526 HzFIDRES 3.362818 HzAQ 0.1488800 secRG 64DW 145.200 usecDE 6.00 usecTE 288.0 KD0 0.00000300 secD1 1.60000002 secD8 0.40000001 secD16 0.00010000 secd20 0.19890000 secIN0 0.00014520 sec
============ CHANNEL f1 =============NUC1 1HP1 6.00 usecP2 12.00 usecPL1 -4.00 dBSFO1 500.1326379 MHz
============ GRADIENT CHANNEL ========GPNAM1 sine.100GPNAM2 sine.100GPX1 0.00 %GPX2 0.00 %GPY1 0.00 %GPY2 0.00 %GPZ1 40.00 %GPZ2 -40.00 %P16 1000.00 usec
F1 - Acquisition parametersND0 2TD 256SFO1 500.1326 MHzFIDRES 13.451274 HzSW 6.885 ppmFnMODE undefined
F2 - Processing parametersSI 1024SF 500.1300144 MHzWDW SINESSB 2LB 0.00 HzGB 0PC 0.20
F1 - Processing parametersSI 1024MC2 TPPISF 500.1300144 MHzWDW SINESSB 2LB 0.00 HzGB 0
N
N N
N
O
O
CH3
CH3
H3C
H
1
3
5
2
8
76
4
3-CH3, 5-H3-CH3, 5-H
Gradient NOESYGradient NOESY
NOESY NOESY gives sequential gives sequential
assignment of peptidesassignment of peptides
23
12 3
CH2
CH2N C
H
N
O
H
C C
H
CH2O
N
H
C
CH2
C
H
OH
O
1 321
Spin-lock
90°
t1 t2
2D ROESY pulse program2D ROESY pulse program
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.
mixing timemixing time
Structure Determination ProceduresStructure Determination Procedures
1D 1D 11H & H & 1313C & DEPT (+MSC & DEPT (+MS 、、 IRIR ,, basic chemical structure basic chemical structure or or functional groups informationfunctional groups information ))Establish Establish 1313C-C-11H connections by thru bond JH connections by thru bond JCHCH couplin couplingsgs HMQCHMQC 、、 HSQCHSQC 、、 HSQC-TOCSY experiments HSQC-TOCSY experiments Establish Establish 11H-H-11H connection (spin systems or partial piH connection (spin systems or partial pieceseces )) Decoupled Decoupled 11H, 1D TOCSY, 2D H, 1D TOCSY, 2D 11H-H-11H COSY, TOCSY expH COSY, TOCSY expts.ts. (( usually starts with well-resolved usually starts with well-resolved 11H signalsH signals ))Long range connectionsLong range connections (( connecting spin systems connecting spin systems & assigning & assigning quaternary carbonquaternary carbon )) 1D NOESY & 2D HMBC, NOESY, ROESY experiments1D NOESY & 2D HMBC, NOESY, ROESY experiments3D structure or conformation determination3D structure or conformation determination 1D NOESY & 2D NOESY, ROESY, (HSQC)-NOESY expts.1D NOESY & 2D NOESY, ROESY, (HSQC)-NOESY expts.