C8953
NMR structural analysis - seminar
1H-13C APT + 2D NMR spectra, COSY
Jan Novotný
novotnyjan@mail.muni.cz
March 11, 2020
1
H NMR spectrum of naringenine in d6-acetone
1
H NMR spectrum of naringenine in d6-acetone
1
2
3
5a,b
4
6
8
9
7
9
1
7
2
3
6
8
4
5a,b
Basics of 1D FT spectroscopy
Draw FT representation of attached FID
records (reciever is located in the +y
direction):
Basics of 1D FT spectroscopy
Draw FT representation of attached FID
records (reciever is located in the +y
direction):
Heteronuclear spin echo
By using vector diagrams determine the result of attached pulse sequence.
Lets consider isolated spin systems a) 13C-1H and
b) 13C-1H2.
τ=1/2J
90y 180x
13
C
τ=1/2J
180x
1
H
CPD
Heteronuclear spin echo
By using vector diagrams determine the result of attached pulse sequence.
Lets consider isolated spin systems a) 13C-1H and
b) 13C-1H2.
τ=1/2J
90y 180x
13
C
τ=1/2J
1
H
CPD
x
y
z
CH...J=JHC
CH2...J=2*JHC
y
x
y
z
y
180x
y
y
-Mx
+Mx
CH2ββCH2αα
CH2αα
CH2αα
CH2αα CH2ββ
CH2ββ
CH2ββ
CHβ CHβ
CHβ
CHβ
CHα
CHα
CHα
CHα
APT - Attached Proton Test
based on heteronuclear spin echo
t1 = 1/1JCH
phase of signals distinguished according to numbers of
directly attached protons
Cq, CH2 positive
CH, CH3 negative
Different 1
JCH =⇒ different intesities
90x 180y
180
C-13 acq
dec
t1
-1
-0.5
0
0.5
1
0 0.5 1 1.5 2
Relativeintensity
t (1/1
J )
Cq
CH
CH2
CH3
13
C APT Nicotine
13
C APT Nicotine
11
10
9
127
3
2,6
54
13
C APT 4
13
C APT 4
2D NMR
Second dimension f1
preparation period
=⇒ coherence
evolution period
t1
FT
−→ f1
increments
evolution of
coherence
mixing period
transfer of
encoded
magnetisation
measurable
signal
detection of signal
t2
FT
−→ f2
acq
t1
prep
evolution
mixing
detection
t2
5t1
4t1
3t1
2t1
t1
2D NMR
2D spektrum
FT in t1 - modulated 1D spectra
FT in t2 - 2D spectrum
COSY
easiest 2D
experiment
correlates H nuclei
based on 2/3J
coupling
through 2, 3, (4)
bonds
antiphase
off-diagonal
crosspeak between
coupled atoms
DQF-COSY modification
of basic
sequence, diagonal
crosspeaks in
absorption phase
acq
t1 t2
))
90 9090
P E M D
f1
f2
f1
f2
Hints for beginners
Determination of individual spin systems - sharing
off-diagonal crosspeaks
Isolated protons - only diagonal crosspeak
Already known rules: symmetry, diastereotopicity, most
shielded/deshielded atoms etc.
COSY : β-cyclodextrine
5.0
5.0
4.5
4.5
4.0
4.0
3.5
3.5
δ2 - 1
H (ppm)
5.0 5.0
4.5 4.5
4.0 4.0
3.5 3.5
δ1-1
H(ppm)
COSY : β-cyclodextrine
5.0
5.0
4.5
4.5
4.0
4.0
3.5
3.5
δ2 - 1
H (ppm)
5.0 5.0
4.5 4.5
4.0 4.0
3.5 3.5
δ1-1
H(ppm)
H1'-H1'
H1'-H2' H2'-H2'H3'-H2'
H3'-H4'
H3'-H3'
H4'-H4'
H5'-H4'
H6'-H5'
Chinin - 1D 1
H
COSY - Chinin
10
10
8
8
6
6
4
4
2
2
δ2 - 1H (ppm)
8 8
6 6
4 4
2 2
δ1-1H(ppm)
5
10
6
9
7
8
3
2
4
N
1
O
CH3
22
11
12
OH
17
16
N
13
15
14
18
19
20
21
COSY - Chinin10 8 6 4 2
8 8
6 6
4 4
2 2
δ1-1H(ppm)
5
10
6
9
7
8
3
2
4
N
1
O
CH3
22
11
12
OH
17
16
N
13
15
14
18
19
20
21
COSY - Chinin
10
10
8
8
6
6
4
4
2
2
δ2 - 1H (ppm)
8 8
6 6
4 4
2 2
δ1-1H(ppm)
5
10
6
9
7
8
3
2
4
N
1
O
CH3
22
11
12
OH
17
16
N
13
15
14
18
19
20
21
21
15,17
14,18
COSY - Chinin
10
10
8
8
6
6
4
4
2
2
δ2 - 1H (ppm)
8 8
6 6
4 4
2 2
δ1-1H(ppm)
5
10
6
9
7
8
3
2
4
N
1
O
CH3
22
11
12
OH
17
16
N
13
15
14
18
19
20
21
21
15,17
14,18
H20-H21'/H21''
H20-H19
H19-H18'/18''
H14'/''-H15'/''
COSY - Chinin
10
10
8
8
6
6
4
4
2
2
δ2 - 1H (ppm)
8 8
6 6
4 4
2 2
δ1-1H(ppm)
5
10
6
9
7
8
3
2
4
N
1
O
CH3
22
11
12
OH
17
16
N
13
15
14
18
19
20
21
15,17
H12-H17'/''
H11-H12
H3-H11
H2-H3
COSY - Chinin
10
10
8
8
6
6
4
4
2
2
δ2 - 1H (ppm)
8 8
6 6
4 4
2 2
δ1-1H(ppm)
5
10
6
9
7
8
3
2
4
N
1
O
CH3
22
11
12
OH
17
16
N
13
15
14
18
19
20
21
H8-H6
H9-H8
NOESY-600ms
H6-H11
H8-H22 H6-H22
H3-H17
H2
H3
H9
H8
H6
H11
H11
H12
H17''
H17'
H21
H20
H19
H18'
H18''
H14''
H15''
H15'
H22
H16-H17'
H19-H16
Next topic
1H-1H correlations (NOESY, ROESY, TOCSY)