common nmr method for structure elucidation of organic molecule
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Structure Elucidation with NuclearStructure Elucidation with Nuclear
Magnetic Resonance (NMR)Magnetic Resonance (NMR)
This was prepared from any material downloaded from internet.
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NMRspectrum Structure
Chem. ShiftJ CouplingPeak Int.
NOECorrelationH HC H O
OOH
O
CH3
O
H
OH
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
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Connections through spaceConnections through spacedipolar couplingdipolar coupling1D1D2D NOESY2D NOESYROESY, HOESY(HSQCROESY, HOESY(HSQC--NOESY)NOESY)usageusage connecting spin systemsconnecting spin systems structure determinationstructure determination
Connections through bondsConnections through bondsspinspin--spin couplingspin couplingHomonuclearHomonuclear 1D1D2D2D 11HH--11H COSY, DQFH COSY, DQF--COSY, TOCSYCOSY, TOCSY
usageusagespin system assignmentspin system assignmentHeteronuclearHeteronuclear
Direct (detectDirect (detect 1313C):C): APT, DEPT, HETCORAPT, DEPT, HETCOR
Inverse (detectInverse (detect11H):H): HMQC, HSQC, HMBC, HSQCHMQC, HSQC, HMBC, HSQC--COSY,COSY,
HSQCHSQC--TOCSY, HMQCTOCSY, HMQC--TOCSYTOCSY
usageusageassigningassigning heteronucleiheteronucleiconnecting spin systemsconnecting spin systems
Some common NMR experiments:Some common NMR experiments:
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COSY spectrum is used for determining the connectivities
between protons on the basis of geminal and vicinal couplings.Disadvantage: bulky dispersive diagonal peaks.
C C H
HH
Vicinal
Geminal
2D COSY2D COSY HomonuclearHomonuclear ShiftShift COCOrrelationrrelation SSpectroscoppectroscopYY
t1 AQ
The basic COSY (x=45 or 90) pulse sequence
90 xt2
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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_ 20001102
Time 8.04INSTRUM DRX500PROBHD 5 mm TBI 1H/PULPROG cosygpTD 1024SOLVENT CDCl3
NS 1DS 16SWH 2185.315 HzFIDRES 2.134096 Hz
AQ 0.2345700 secRG 40.3DW 228.800 usecDE 6.00 usecTE 288.0 K
D0 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.100
GPNAM2 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 MHz
WDW QSINESSB 0LB 0.00 HzGB 0PC 0.20
F1 - Processing parametersSI 1024
MC2 QFSF 500.1300144 MHz
WDW QSINE
SSB 0LB 0.00 HzGB 0
11 22 33 44
11
22
33
44
HO C
H1
H
C C C
H2
H
H3
H
H4
H
H
11--2222--33
33--44
2D Gradient COSY2D Gradient COSY--4545
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2D2D DQFDQFCOSYCOSY((DDoubleouble--QQuantumuantum FFilterediltered COSYCOSY))
90 90 90
t
1 t
2
allow J coupling constant measurementsallow J coupling constant measurements
observing crossobserving cross--peaks close to diagonalpeaks close to diagonal
But lower sensitivity (~40%) of COSY expt.But lower sensitivity (~40%) of COSY expt.
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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_ 20001103
Time 4.01INSTRUM DRX500PROBHD 5 mm TBI 1H/PULPROG cosydfetgp.2TD 4096SOLVENT Pyr
NS 8DS 16SWH 3822.630 HzFIDRES 0.933259 Hz
AQ 0.5359376 secRG 512DW 130.800 usecDE 6.00 usecTE 288.0 KD0 0.00000300 secD1 1.60000002 sec
D11 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.100
GPNAM2 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 MHz
FIDRES 7.465717 HzSW 7.643 ppmFnMODE undefined
F2 - Processing parametersSI 4096SF 500.1300038 MHz
WDW SINESSB 2LB 0.00 HzGB 0PC 0.20
F1 - Processing parametersSI 1024
MC2 echo-antiechoSF 500.1300038 MHz
WDW SINESSB 2LB 0.00 HzGB 0
2D Gradient DQF2D Gradient DQF--COSYCOSY
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33J=8.00HzJ=8.00Hz
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TOCSYTOCSY ((TOTOtaltal CCorrelationorrelation SSpectroscoppectroscopYY) or) or
HOHAHAHOHAHA((HoHomonuclearmonuclear HaHartmanrtman--HaHahn Spectroscopy)hn Spectroscopy)
t1 MLEV17 AQ
Pulse sequence for a TOCSY spectrum.
Different mixing time gives different degree of relay ofDifferent mixing time gives different degree of relay of
correlation. At small mixing time, TOCSY spectrum is similar tocorrelation. At small mixing time, TOCSY spectrum is similar toCOSY 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
H
HO C
H1
H
C C C
H2
H
H3
H
H4
H
H
COSYCOSY RLRL--COSYCOSY TOCSYTOCSY
mixing timemixing timet2
2D G di TOCSY2D G di t TOCSY
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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 DRX500
PROBHD 5 mm TBI 1H/PULPROG mlevetgpTD 1024SOLVENT CDCl3
NS 8DS 16SWH 2185.315 HzFIDRES 2.134096 Hz
AQ 0.2345700 secRG 16DW 228.800 usecDE 6.00 usecTE 300.0 KD0 0.00000300 secD1 1.79999995 secD9 0.01000000 secD11 0.03000000 sec
D16 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 dB
PL10 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 MHz
WDW SINESSB 3LB 0.00 HzGB 0PC 0.20
F1 - Processing parametersSI 1024
MC2 echo-antiechoSF 500.1300144 MHz
WDW 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 44
11
22
33
44
11--2222--33
33--44
2D G di t TOCSY2D G di t TOCSY
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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 DRX500
PROBHD 5 mm TBI 1H/PULPROG mlevetgpTD 1024SOLVENT CDCl3
NS 8DS 16SWH 2185.315 HzFIDRES 2.134096 Hz
AQ 0.2345700 secRG 16DW 228.800 usecDE 6.00 usecTE 300.0 KD0 0.00000300 secD1 1.79999995 secD9 0.02000000 secD11 0.03000000 sec
D16 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 dB
PL10 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 MHz
WDW SINESSB 2LB 0.00 HzGB 0PC 0.20
F1 - Processing parametersSI 1024
MC2 echo-antiechoSF 500.1300144 MHz
WDW SINESSB 2LB 0.00 HzGB 0
2D Gradient TOCSY,2D Gradient TOCSY, mixing time=20ms11 22 33 44
11
22
33
44
11--22
22--33
33--44
11--33
22--44
Same as RLSame as RLCOSYCOSYHO C
H1
H
C C C
H2
H
H3
H
H4
H
H
2D G di t TOCSY2D G di t TOCSY
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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 CDCl3
NS 8DS 16SWH 2185.315 HzFIDRES 2.134096 Hz
AQ 0.2343412 secRG 18DW 228.800 usecDE 6.00 usecTE 300.0 KD0 0.00000300 secD1 1.79999995 secD9 0.05000000 secD11 0.03000000 sec
D16 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 256
SFO1 500.1311 MHzFIDRES 8.535453 HzSW 4.369 ppm
F2 - Processing parametersSI 1024SF 500.1300144 MHz
WDW QSINESSB 4LB 0.00 HzGB 0PC 0.20
F1 - Processing parametersSI 1024
MC2 echo-antiechoSF 500.1300144 MHz
WDW QSINESSB 4LB 0.00 Hz
GB 0
2D Gradient TOCSY,2D Gradient TOCSY, mixing time=50ms11 22 33 44
11
22
33
44
11--22
22--33
33--44
11--33
22--4411--44
HO C
H1
H
C C C
H2
H
H3
H
H4
H
H
Total CorrelationTotal Correlation
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DEPT: Distortionless Enhancement by Polarization Transfer
Heteronuclear expt.Detection: 13C
Distinguish
CH, CH2, CH3
By suitable combination of
=45, 90 & 135 spectra
All CHs
Only CH
CH & CH3up
CH2 down
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HETCORHETCOR ((HetHeteronucleareronuclear chemical shiftchemical shift corcorrelation,relation, 11HH -- 1313C COSY)C COSY)
13C
1H t1 1 2
The standard pulse sequence forThe standard pulse sequence for 1313CC--detecteddetected11HH--1313C chemical shift correlation.C chemical shift correlation.
AQt2
1H decoupling
Removing 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
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HSQCHSQC ((HHeteronucleareteronuclear SSingleingle QQuantumuantum CCoherence)oherence)
AQ
1H
13C /2 /2 GARP-1t1 /2 /2
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 easierto perform.
13C decouplingRemoving JCH splittings
HMQCHMQC ((HHeteronucleareteronuclear MMultipleultiple QQuantumuantum CCoherence)oherence)
t2
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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 DRX500
PROBHD 5 mm TBI 1H/PULPROG invietgpsiTD 1024SOLVENT CDCl3
NS 1DS 16SWH 2185.315 HzFIDRES 2.134096 Hz
AQ 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 sec
D16 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 garp
NUC2 13CP3 17.70 usec
P4 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 MHz
WDW SINESSB 2LB 0.00 HzGB 0PC 0.20
F1 - Processing parametersSI 1024
MC2 echo-antiecho
SF 125.7577969 MHzWDW SINESSB 2LB 0.00 HzGB 0
HO C
H1
HC C C
H2
H
H3
H
H4
HH
11 22 33 44
11
22
33
44
HSQC spectrum: HHSQC spectrum: H--C correlatedC correlated
(HMQC and HETCOR spectra look similar)(HMQC and HETCOR spectra look similar)
C-d
imension
HH--dimensiondimension
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HSQCHSQC--TOCSYTOCSY
AQ
1H
13C /2 /2 GARP-1t1 /2 /2
Pulse sequence forPulse sequence for 11HH--detected HSQCdetected HSQC--TOCSY experimentTOCSY experimentthrough singlethrough single--quantum coherence.quantum coherence.
Longer mixing time gives longer extend of spinLonger mixing time gives longer extend of spin--spinspincorrelations.correlations.
SLy
MLEV17
SLy
mixing timemixing time
G di HSQCG di HSQC TOCSY( i i i 10 )TOCSY( i i i 10 )
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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 CDCl3
NS 4DS 16SWH 2185.315 HzFIDRES 2.134096 Hz
AQ 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 sec
D13 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 garp
NUC2 13CP3 15.70 usecP4 31.40 usecPCPD2 89.00 usecPL2 -2.00 dBPL12 13.00 dB
SFO2 125.7633722 MHz
============ GRADIENT CHANNEL ========P16 1000.00 usec
F1 - Acquisition parametersND0 2TD 256
SFO1 125.7634 MHzFIDRES 54.253471 HzSW 110.437 ppm
F2 - Processing parametersSI 1024SF 500.1300144 MHz
WDW SINESSB 2LB 0.00 HzGB 0PC 0.20
F1 - Processing parametersSI 512
MC2 echo-antiechoSF 125.7577961 MHz
WDW SINE
SSB 2LB 0.00 HzGB 0
HO C
H1
H
C C C
H2
H
H3
H
H4
H
H
Same asSame as HSQCHSQCCOSYCOSY spectrumspectrum
Gradient HSQCGradient HSQC--TOCSY(mixing time=10ms)TOCSY(mixing time=10ms)
11 22 33 44
11
22
33
44
11--11
11--22
22--11
HH--dimensiondimension
C-d
imension
G di t HSQCG di t HSQC TOCSY( i i ti 20 )TOCSY( i i ti 20 )
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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 CDCl3
NS 4DS 16SWH 2185.315 HzFIDRES 2.134096 Hz
AQ 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 sec
D13 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 usec
P17 2500.00 usecP28 2000.00 usecPL1 -4.00 dB
PL10 10.00 dBSFO1 500.1310815 MHz
============ CHANNEL f2 =============CPDPRG2 garp
NUC2 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 MHz
WDW SINESSB 2LB 0.00 HzGB 0PC 0.20
F1 - Processing parametersSI 512
MC2 echo-antiechoSF 125.7577961 MHz
WDW SINE
SSB 2LB 0.00 HzGB 0
11 22 33 44
11
22
33
44
HO C
H1
H
C C C
H2
H
H3
H
H4
H
H
Gradient HSQCGradient HSQC--TOCSY(mixing time=20ms)TOCSY(mixing time=20ms)
11--22
22--1111--11
11--33
33--11
C-d
imension
HH
--dimensiondimension
G di t HSQCG di t HSQC TOCSY( i i ti 80 )TOCSY( i i ti 80 )
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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 CDCl3
NS 4DS 16SWH 2185.315 HzFIDRES 2.134096 Hz
AQ 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 sec
D13 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 usec
P17 2500.00 usecP28 2000.00 usecPL1 -4.00 dBPL10 10.00 dBSFO1 500.1310815 MHz
============ CHANNEL f2 =============CPDPRG2 garp
NUC2 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 256
SFO1 125.7634 MHzFIDRES 54.253471 HzSW 110.437 ppm
F2 - Processing parametersSI 1024SF 500.1300133 MHz
WDW SINESSB 2LB 0.00 HzGB 0PC 0.20
F1 - Processing parametersSI 512
MC2 echo-antiechoSF 125.7577961 MHz
WDW SINE
SSB 2LB 0.00 HzGB 0
HO C
H1
H
C C C
H2
H
H3
H
H4
H
H
11 22 33 44
11
22
33
44
Gradient HSQCGradient HSQC--TOCSY(mixing time=80ms)TOCSY(mixing time=80ms)
11--22
22--1111--11
11--33
33--11
11--44
44--11
C-d
imension
HH
--dimensiondimension
HMBCHMBC ((HH t leteronuclear MM lti lultiple BB dond CC l ti S t )orrelation Spectroscopy)
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HMBCHMBC ((HHeteronucleareteronuclear MMultipleultiple--BBondond CCorrelation Spectroscopy)orrelation Spectroscopy)
13
C
1H t1AQ
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
Gradient HMBCGradient HMBC
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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 CDCl3
NS 4DS 16SWH 3443.526 HzFIDRES 3.362818 Hz
AQ 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 sec
IN0 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.100
GPNAM3 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 Hz
SW 167.039 ppmFnMODE undefined
F2 - Processing parametersSI 1024SF 500.1300144 MHz
WDW QSINESSB 0LB 0.00 HzGB 0PC 0.20
F1 - Processing parametersSI 512
MC2 QFSF 125.7577961 MHz
WDW QSINE
SSB 0LB 0.00 HzGB 0
N
N N
N
O
O
CH3
CH3
H3C
H
1
3
5
2
8
76
4
Gradient HMBCGradient HMBC
11
2233
55
11
44
33
22
55
66
77
88
33--44
33--5522--66
22--77
11--77
11--88
HH
--dimensiondimension
C-dimens
ion
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2D2D NOESYNOESY ((NNuclearuclear OOverhauserverhauser EEnhancementsnhancements SSpectroscoppectroscopYY))
t1 m AQ
The NOESY pulse sequence.
C ~ C
Ha HbVC*r-6, r
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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_ 20001103
Time 8.53INSTRUM DRX500PROBHD 5 mm TBI 1H/PULPROG noesygptpTD 1024SOLVENT CDCl3
NS 8DS 16SWH 3443.526 HzFIDRES 3.362818 Hz
AQ 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.100
GPNAM2 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 ppm
FnMODE undefined
F2 - Processing parametersSI 1024SF 500.1300144 MHz
WDW SINESSB 2LB 0.00 HzGB 0PC 0.20
F1 - Processing parametersSI 1024
MC2 TPPISF 500.1300144 MHz
WDW SINE
SSB 2LB 0.00 HzGB 0
N
N N
N
O
O
CH3
CH3
H3CH
1
3
5
2
8
76
4
33--CH3, 5CH3, 5--HH
Gradient NOESYGradient NOESY
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NOESYNOESY
gives sequentialgives sequential
assignment of peptidesassignment of peptides
2
3
1
2 3
CH2
C2N C
H
N
O
H
C C
H
CH2O
N
H
C
CH2
C
H
OH
O
1 321
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Spin-lock
90
t1
t2
mixing timemixing time
2D ROESY pulse program2D ROESY pulse program
For small molecule NOE can be very small or zeroROESY can be used in place of NOESY experiment. ROE
intensity is also related to the H-H distances.
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Structure Determination ProceduresStructure Determination Procedures
1D1D 11H &H & 1313C & DEPT (+MSC & DEPT (+MSIRIRbasic chemical structure orbasic chemical structure orfunctional groups informationfunctional groups information
EstablishEstablish
1313
CC--
11
H connections by thru bond JH connections by thru bond JCHCH couplingscouplingsHMQCHMQCHSQCHSQCHSQCHSQC--TOCSY experimentsTOCSY experiments
EstablishEstablish 11HH--11H connection (spin systems or partial piecesH connection (spin systems or partial piecesDecoupledDecoupled 11
H, 1D TOCSY, 2DH, 1D TOCSY, 2D 11
HH
--11
H COSY, TOCSYH COSY, TOCSY
exptsexpts
.
.
usually starts with wellusually starts with well--resolvedresolved 11H signalsH signalsLong range connectionsLong range connectionsconnecting spin systems & assigningconnecting spin systems & assigning
quaternary carbonquaternary carbon1D NOESY & 2D HMBC, NOESY, ROESY experiments1D NOESY & 2D HMBC, NOESY, ROESY experiments
3D structure or conformation determination3D structure or conformation determination
1D NOESY & 2D NOESY, ROESY, (HSQC)1D NOESY & 2D NOESY, ROESY, (HSQC)--NOESYNOESY exptsexpts..
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