utilización combinada de experimentos de rmn mw: 324,42

1

Utilización combinada de experimentos de RMN

UNIVERSITAT DE BARCELONA

ServeisServeis CientCientííficfic TTèècnicscnics

Varian Mercury 400

Unitat de RMN dUnitat de RMN d’’Alt CampAlt Camp Quinina

C20H24N2O2

MW: 324,42

http://en.wikipedia.org/wiki/Quinine

Unitat de RMN dUnitat de RMN d’’Alt CampAlt Camp Experimentos básicos I

1D 1H1D 1H--RMNRMNDet. NDet. Núúmero de protones, desplazamientos mero de protones, desplazamientos

ququíímicos, acoplamientos 1Hmicos, acoplamientos 1H--1H1H1D1D--13C13C--RMN RMN

•• Det. NDet. Núúmero de carbonos no equivalentesmero de carbonos no equivalentes•• Grupos funcionalesGrupos funcionales

Experimentos 1D con otros nExperimentos 1D con otros núúcleoscleos•• ( ( 31P, 19F, 31P, 15N31P, 19F, 31P, 15N…………))

gHSQCgHSQCDeterminaciDeterminacióón de las correlaciones directasn de las correlaciones directas 1H1H--13C13C

NO

Unitat de RMN dUnitat de RMN d’’Alt CampAlt Camp Experimentos básicos II

TOCSYTOCSY

Localización de los sistemas de acoplamiento.

COSY / DQFCOSYCOSY / DQFCOSY

Determinación de los acoplamientos directos 1H-1H ( 2-3 enlaces)

HMBCHMBCCorrelaciones a larga distancia 1H y X

NOESY/ROESYNOESY/ROESY

Relaciones en el espacio, det estructura 3D

2

Unitat de RMN dUnitat de RMN d’’Alt CampAlt Camp

ppm (f1)1.02.03.04.05.06.07.08.09.0

1.00 0.97 0.990.910.90 0.980.89 1.86 0.92 2.84 1.00 1.94 1.88 0.94 2.811.91

1D- H1

ppm (f1)4.905.005.105.205.305.405.505.605.70

Unitat de RMN dUnitat de RMN d’’Alt CampAlt Camp Espectro C13

ppm (t1)050100150

ppm (t1)145.0150.0155.0

At=1.1 s, d1=1s nt=30001h 50 minutos

Unitat de RMN dUnitat de RMN d’’Alt CampAlt Camp DEPT

ppm (t1) 255075100125150

nt=1024 d1=1 at=1.2 mult=1,1.5

~38 minutos

Unitat de RMN dUnitat de RMN d’’Alt CampAlt Camp Información previa

Desplazamientos quDesplazamientos quíímicos micos 11HHIntegraciónDeterminación de constantes de acoplamiento

24 protones

Desplazamientos quDesplazamientos quíímicos micos 1313CC

20 Carbonos zona aromzona aromááticaticaalifalifááticosticos

Posibles solapamientos

3


157,67147,66147,43144,09141,66131,40126,57121,43118,44114,43101,3071,7459,9656,9055,7043,2039,8327,8027,5121,72

Tablas de desplazamientos químicos

20 Carbonos20 Carbonos

23+1 protones23+1 protones

(4,7 Hz)(9,2 Hz)(4,7 Hz)(2,7 ; 9,2 Hz)(2,7 Hz)(7,6 , 10,3 16.9 Hz)(3,8 Hz) (1,4 (t) 16.9Hz)(1,2(t) 10,3 Hz) *

(10,1 13,7 Hz)comp

8,55 (1) d7,92 (1) d7,47 (1) d7,29 (1) dd7,27 (1) d5.71 (1) m5,53 (1) d4,95 (1) dt4,92 (1) dt4,37 (*) b 3,86 (3) s3.46 (1) mb3,12 (1) mb3,07 (1) dd2,65 (2) mc2,26 (1) mc1,77 (2) mc1,72 (1) mc1,51 (2) comp

*


4.94.955.005.055.105.155.205.255.305.355.405.455.505.555.605.655.705.755.80f1 (ppm)

5.75 (ddd, J = 17.53, 10.32, 7.53 Hz, 1H), 4.98 (td, J = 17.25, 1.32, 1.32 Hz, 1H), 4.94 (td, J = 10.45, 1.25, 1.25 Hz, 1H)

Incremento de resolución

Procesado Perdida de sensibilidad


ppm

(t1

)50

100

150

ppm (t2)1.001.502.002.503.003.504.004.505.005.506.006.507.007.508.008.50

25

50

75

100

125

150ppm (t1

ppm (t1)1.02.03.04.05.06.07.08.0

CH2

CH

gHSQC editado

Nt =2 ni=256 Tiempo=30 min

Determinación de correlaciones directas 1H-X

Discriminación CH2 vs CH y CH3

C C C C

H H H H

CDCl3


ppm (t2)4.755.005.255.505.756.006.256.506.757.007.257.507.758.008.258.50

95.0

100.0

105.0

110.0

115.0

120.0

125.0

130.0

135.0

140.0

145.0

150.0ppm (t1pp

m (

t1)

100

110

120

130

140

150

p p m ( t 1 )5 . 0 05 . 5 06 . 0 06 . 5 07 . 0 07 . 5 08 . 0 08 . 5 0

CH2

gHSQC-I

4


ppm (t2)1.501.752.002.252.502.753.003.253.50

20.0

25.0

30.0

35.0

40.0

45.0

50.0

55.0

60.0

65.0

ppm (t1ppm

(t1)

2030

4050

60

CH2

CH2

CH

gHSQC-II

ppm (t1)1.502.002.503.003.50

Unitat de RMN dUnitat de RMN d’’Alt CampAlt Camp gHSQC-III

4 carbonos cuaternarios10 CH5 CH21 CH3

20 Carbonos 20 Carbonos

ppm (t2)3.754.004.254.504.755.005.255.50

55.0

57.5

60.0

62.5

65.0

67.5

70.0

72.5ppm (t1

pm (t1)4.004.505.005.50

OCH3

Unitat de RMN dUnitat de RMN d’’Alt CampAlt Camp gHSQCAD

1.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5f2 (ppm)

20

30

40

50

60

70

80

90

100

110

120

130

140

150

F2 campana seno 90F2 campana seno 90F1 seno cuadrado 90F1 seno cuadrado 90

F1

Unitat de RMN dUnitat de RMN d’’Alt CampAlt Camp Listado de señales

1.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.0f2 (ppm)

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

f1 (

ppm

)

{8.70,147.83}

{5.74,141.76}

{8.00,131.93} {7.98,131.92}

{7.33,121.78}

{7.32,121.80}

{7.53,118.64}

{5.00,114.77} {4.96,114.77}

{4.93,114.77}

{7.24,101.35}

{5.62,71.92}

{3.18,60.03}{3.15,56.97} {3.13,56.94}

{3.10,56.96}

{3.89,55.85}

{3.50,43.29} {2.71,43.26}

{2.31,39.80}

{1.85,27.84} {1.76,27.47}

{1.53,27.48}

{1.58,21.90}

Combinación de multipletes

5

Unitat de RMN dUnitat de RMN d’’Alt CampAlt Camp Mnova Tabla de señales

47.64133.55-8.81.7421.9121

47.64133.55-7.81.5221.9120

47.64133.55-9.31.4927.7119

39.70166.93-9.81.7327.7418

31.76166.9316.71.8128.0217

47.64166.9314.92.2740.1216

55.58133.55-14.42.6543.5015

55.58133.55-8.93.4743.5014

23.82233.719.23.8656.0113

38.20133.93-4.83.0857.2112

37.17134.02-12.02.6557.2111

47.64166.9316.13.1260.2710

39.70133.5521.55.5472.099

31.76233.712.07.22101.748

15.88166.93-10.24.98114.907

31.76166.93-13.04.91114.916

23.82133.5521.77.48118.945

31.76133.5512.17.29121.944

31.76133.5516.17.93131.943

15.8866.773.85.70142.202

15.88166.932.48.56147.951

Ancho 2Ancho 1Intensidadf2 (ppm)f1 (ppm)

EXCEL. WORD ...........


1H 1H13C 13C

Cuaternarios

8,55 (1) d7,92 (1) d7,47 (1) d7,29 (1) dd7,27 (1) d5.71 (1) m5,53 (1) d4,95 (1) dt4,37 (*) b3,86 (3) s3.47 (1) mb

3,12 (1) mb3,07 (1) dd2,26 (1) mc1,77 (1) mc1,76 (1) mc1,75 (1) mc

147,43131,40118,44121,43101,30141,6671,74

114,43a

55,7043,20 b

157,67147,66144,09126,57

59,9656,9039,8327,8027,5121,72

4,92 (1)

2,65 (1)

2,65 (1)

1,50 (1)1,51 (1)

OH

OCH3

CH2

CH2

CH2

CH2

Resumen, 1H-13C


C20H24N2O2

MW: 324,42

1

2

N

HHO

N

H3CO

H

H1 6

5

4

3

2

7

89

4'5'

6'

1'

7'8'

10'

9'

2'

3'

10

11

Unitat de RMN dUnitat de RMN d’’Alt CampAlt Camp DQFCOSY /COSY

ppm (t2)2.03.04.05.06.07.08.0

2.0

3.0

4.0

5.0

6.0

7.0

8.0

ppm (t1

Nt=2 ni=256Tiempo=15 min

Determinación de acoplamientos 1H-1H directos C C C C

H H H H

H

6

Unitat de RMN dUnitat de RMN d’’Alt CampAlt Camp Procesado

PredicciPrediccióón lineal n lineal Ajuste de la funciAjuste de la funcióón de procesadon de procesado

Ajuste de Fase (exp en fase: Noesy, hsqc, tocsy, etc)

ReducciReduccióón ruido en t1n ruido en t1SimetrizaciSimetrizacióónn ((expexp CosyCosy))

Unitat de RMN dUnitat de RMN d’’Alt CampAlt Camp Experimentos en AV

F2 Seno cuadrado 0 F2 Seno cuadrado 0 F1 Seno cuadrado 0F1 Seno cuadrado 0

Seno cuadrado 0º Campana de seno 0º

7.07.17.27.37.47.57.67.77.87.98.08.18.28.38.48.58.68.78.88.99.09.1f2 (ppm)

7.0

7.1

7.2

7.3

7.4

7.5

7.6

7.7

7.8

7.9

8.0

8.1

8.2

8.3

8.4

8.5

8.6

8.7

8.8

8.9

9.0

9.1

6.97.07.17.27.37.47.57.67.77.87.98.08.18.28.38.48.58.68.78.88.99.0f2 (ppm)

6.8

6.9

7.0

7.1

7.2

7.3

7.4

7.5

7.6

7.7

7.8

7.9

8.0

8.1

8.2

8.3

8.4

8.5

8.6

8.7

8.8

8.9

9.0

9.1

9.2

Unitat de RMN dUnitat de RMN d’’Alt CampAlt Camp Posibilidades de mejora de la resolución en t2

Una alternativa a la Predicción lineal ?

Llenado de ceros

Predicción lineal

Covariance

(http://spin.magnet.fsu.edu/software/covNMR/covNMR.html)

Posibles artefactos•Señales similares al Tocsy•Señales fantasma

Unitat de RMN dUnitat de RMN d’’Alt CampAlt Camp Mejora de la resolución Covariance-NMR

7

Unitat de RMN dUnitat de RMN d’’Alt CampAlt CampgCOSY-I nt=1 ni=256

Tiempo < 10 min

ppm (t2)7.508.008.50

7.00

7.50

8.00

8.50

ppm (t1

R

R

R

R

7,47

8.55

R

R

R7.27

7.92

7.29

ppm (t1)7.2007.2507.300

7.9007.950

*


5.53, 3.12, 1.76,1.515.53, 3.12, 1.76,1.51

R HHO

RH

HH

R

5.53

3.12

1.76 1.51

ppm (t2)2.03.04.05.06.0

1.0

2.0

3.0

4.0

5.0

6.0ppm (t1

OH

gCOSY-II


OH

5.71, 4,95, 4.925.71, 4,95, 4.92

ppm (t2)2.03.04.05.06.0

1.0

2.0

3.0

4.0

5.0

6.0ppm (t1

H

H

H

RR

H

2.26

5.71

4,954.92

gCOSY-III Unitat de RMN dUnitat de RMN d’’Alt CampAlt Camp

3.07, 2.65, 2.26

2.26

3.072.65

H

H

H

R

H

R

H HH

ppm (t2)1.502.002.503.003.50

1.50

2.00

2.50

3.00

3.50

ppm (t1

gCosy-IV

8


3.47, 2.65, 1.77, 1.70, 1.50

R

R

N

R

R

H

HH

H H

3.47

2.65

1.70

1.50

1.77

ppm (t2)1.502.002.503.003.50

1.50

2.00

2.50

3.00

3.50

ppm (t1

gCOSY-V

2H


ppm (t2)1.02.03.04.05.06.07.08.09.0

1.0

2.0

3.0

4.0

5.0

6.0

7.0

8.0

9.0

ppm (t1

TOCSY

C C C C C C

O

H HH H H

Determinación sistemas de acoplamiento

Quinina 2mg/0.8 ml

TOCSYnt=4 ni=256 mix=0.08tiempo 1h


8.55, 7,47

7.92, 7.29, 7.27

R

R

R

R

R

R

R

7.007.508.008.50

7.00

7.50

8.00

8.50

9.00ppm (t1

Tocsy-I Unitat de RMN dUnitat de RMN d’’Alt CampAlt Camp Tocsy-II

5.71, 4,95, 4.92

5.53, 3.12, 1.76,1.51

ppm (t2)1.502.002.503.003.504.004.505.005.50

2.0

3.0

4.0

5.0

ppm (t1

R HHO

RH

HH

R

5.53

3.12

1.76 1.51

9


3.07, 2.65, 2.26

ppm (t2)1.502.002.503.003.50

1.50

2.00

2.50

3.00

3.50

ppm (t1

3.47, 2.65, 1.77, 1.70, 1.50

2.26

3.072.65

H

H

H

R

H

R

H HH

R

R

N

R

R

H

HH

H H

3.47

2.65

1.70

1.50

1.77

Tocsy-III Unitat de RMN dUnitat de RMN d’’Alt CampAlt Camp Resultado Tocsy

3.47, 2.65, 1.77, 1.70, 1.50

5.71, 4,95, 4.92

5.53, 3.12, 1.76,1.51

8.55, 7,47

7.92, 7.29, 7.27

3.07, 2.65, 2.26

8,55 (1)7,92 (1)7,47 (1)7,29 (1)7,27 (1)5.71 (1)5,53 (1)4,95 (1)4,37 (*)3,86 (3)3.47 (1)3,12 (1)3,07 (1) 2,26 (1)1,77 (1)1,76 (1)1,75 (1)

4,92 (1)

2,65 (1)

2,65 (1)

1,50 (1)1,51 (1)


ppm (t1)2.03.04.05.06.0

OH

OCH3

11, 11’10 9 6 a 6 b2a8 32b5 a 5 b

7 b7 a

4

7.508.008.50N

HHO

N

H3CO

H

H1 6

5

4

3

2

7

89

4'5'

6'

1'

7'8'

10'

9'

2'

3'

10

11

7’

8’8’2’

3’

Asignación 1HUnitat de RMN dUnitat de RMN d’’Alt CampAlt Camp Asignación 13C

1)100110120130140

t1)2030405060

7’2’ 8’ 3’ 5’

87

543

24

OCH3

9

1110

N

HHO

N

H3CO

H

H1 6

5

4

3

2

7

89

4'5'

6'

1'

7'8'

10'

9'

2'

3'

10

11

10


N

HHO

N

H3CO

H

H

147,4

131,4

118,4

121,4

101,3

141,6

71,7

114,4

55,7

43,259,9

56,9

39,8

27,827,521,7

3.472.65

1.701.50

1.77

2.26

3.072.65

5.714,954.92

5.53

3.121.761.51

7,47

8.55

7.27

7.92

7.29


ppm (t2)7.007.508.008.50

100

150

ppm (t1

C C C C

H H H HHSQC

HMBC

gHMBC

Nt =4 ni=256 Jxnh=Tiempo=30 min

5 0 0

ppm (t2)7.007.508.008.50

100

150

ppm (t1

* Ghsqc superpuesto

**

*

*

*


ppm (t2)4.505.005.506.00

50

100

150

ppm (t1

Comparación gHMBC-gHSQC

* Ghsqc superpuesto

pp

m (

t1)

50

10

01

50

50

100

150

ppm (t1

*

*


N

HHO

R

H3CO

HR

H

H

H

H

H

8.55 143.3, 148.3 118.4

7.92 157.4 126.4

8.55

7.92

148.3

7.27

7.27 157.4, 148.3, 143.4, 121.4

126.4

157.4

143.3

HMBC asignación C cuaternarios

7,47 7,47 148.3, 126.4, 71.7

7.29

7.29 157,4 143.4, 101.3

11


N

HHO

N

H3CO

H

H

5.71 27.9, 39.8, 56.9

5.53 21.5, 59.9, 118.6, 148.13

ppm (t2)5.506.00

pp

m (

t1)

50

10

01

50


N

HHO

N

H3CO

H

H

147,4

131,4

118,4

121,4

101,3

141,6

71,7

114,4

55,7

43,259,9

56,9

39,8

27,827,521,7

3.472.65

1.701.50

1.77

2.26

3.072.65

5.714,95

4.92

5.53

3.121.761.51

7,47

8.55

7.27

7.92

7.29

148.3

126.4

157.4

143.3


ppm (t2)2.03.04.05.06.07.08.0

1.0

2.0

3.0

4.0

5.0

6.0

7.0

8.0

ppm (t1

NOESY

Nt =2 ni=256 Mix=0.5 sTiempo=2 horas


ROESYNOESY Mix=0.5 s Mix=0.3 s

ppm (t2)1.502.002.503.003.504.004.505.005.506.006.507.007.508.00

1.50

2.00

2.50

3.00

3.50

4.00

4.50

5.00

5.50

6.00

6.50

7.00

7.50

8.00

8.50ppm (t1

ppm (t2)2.03.04.05.06.07.08.0

2.0

3.0

4.0

5.0

6.0

7.0

8.0

ppm (t1

NOESY vs ROESY

12


Curvas del Noe/Roe en función del tiempo de correlación.

Frecuencia del equipo 500 MHz y distancia entre protones 2.9 Aº.

1

-10.1 10

-0.5

0.5

τc (ns)

η

NOE

ROE

0

68%

100%

NOESY/ ROESY

Noesy

Noe-1d

Roesy

Tiempo de mezcla

ω0 τc=1.12ωτc << 1 ωτc >> 1

pequeñas medianas grandes


ppm (t2)2.03.04.05.06.07.08.0

1.0

2.0

3.0

4.0

5.0

6.0

7.0

8.0

ppm (t1

ppm (t2)2.03.04.05.06.07.08.0

1.0

2.0

3.0

4.0

5.0

6.0

7.0

8.0

ppm (t1

Comparación NOESY COSY

NOESY

COSY


NOESY-I

ppm (t2)7.007.508.008.50

2.0

3.0

4.0

5.0

6.0

7.0

8.0

9.0ppm (t1

N

HHO

N

H3CO

H

H

H

H1.71

3’

5’

3’ 5’

9

88

5

5


N

HHO

N

H3CO

H

H

H

H

H

ppm (t2)1.502.002.503.003.504.004.505.005.50

2.0

3.0

4.0

5.0

ppm (t1

1 . 5 02 . 0 02 . 5 03 . 0 03 . 5 04 . 0 04 . 5 05 . 0 05 . 5 0

3.47

1,51

2.65

NOESY-II

6b 2b91011

OH 6a18 2a

34

5a 7a5b 7b

13


N

HHO

N

H

H3CO

H

H

HH

H H

H

H

2.26

1.77

1.50

3.47

3.07

1.70

N

HHO

N

H

H3CO

H

H

HH

H H

H

H

ppm (t2)1.502.002.503.003.50

1.50

2.00

2.50

3.00

3.50

4.00ppm (t1

2.26

1.77 1.50

3.47

3.07

1.70

6b 2b

46a

18 2a

35a 7a

5b 7b

NOESY-III


ppm (f1) 0.501.001.502.002.503.003.504.004.505.005.506.006.507.007.508.008.50ppm (f1) 0.501.001.502.002.503.003.504.004.505.005.506.006.507.007.508.008.50ppm (f1) 0.501.001.502.002.503.003.504.004.505.005.506.006.507.007.508.008.50

NOESY1D

1.77

1.70

6a

6b

3.47

3

42a1110

5b

2.26

1.503.07

N

HHO

N

H

H3CO

H

H

HH

H H

H

H

5b92’ , 3’

PFGE NOE1Dmix=500 ms nt=128


N

HHO

N

H3CO

H

H

H

HH

H

H

H

H

H

HH

147,4

131,4

118,4

121,4

101,3

141,6

71,7

114,4

55,7

43,259,9

56,9

39,8

27,8

27,521,7

148.3126.4

157.4

143.3

3.47

2.651.70

1.50

1.77

2.26

3.072.65

5.71

4,95

4.92

5.53

3.121.761.51

7,47

8.55

7.27

7.92

7.29

utilización combinada de experimentos de rmn mw: 324,42

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