C8953 NMR strukturní analýza seminář
Identification of an unknown compound Jan Novotný
176003@is.muni.cz
April 13, 2022
Task 0: Classification of an unknown substance
a    k AVUa A
1
1
V   a   /A aA*.. . ,
JAJL
JÜU_UlJ_L
JL-IL.
—^_1*-
-i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—r~
2
Ondřej Jurček
9
1 ^
ßbrV. 26, 2M7 O^Vd*
Task 0: Classification of an unknown substance
Terpene
Carbohydrate
Streroid
Peptide
a    í AAaaA
Alkaloid
1 1    .1 ll.
1
jjjJL
JUNiiii
J/lA
Coumarine
JLU_UlJ_L
t—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—i—r
7
T 5
r
3 2 1 ppm
Ondrej Jurček
ßbrV. 26, 2&7 O^Vd*
Task 1: Ci0Hi3NO
5[ppm]        Multiplicity Integral 1.05             triplet 3 1.75             singlet 3 3.70             quartet 2
7-7.60 complex multiplet 5
Ondřej Jurček
tori'. 26, 2SL7 O^Vd*
Task 1: Ci0Hi3NO
5[ppm]        Multiplicity Integral
1.05 a triplet 3 f A-1\|
1.75 c singlet 3 \,_/J \_
3.70 b quartet 2 e        d //
7-7.60 d-f complex multiplet 5 O
C
/
Ondřej Jurček
tori'. 26, 2SL7 0*Vd*
Task 2: CnHi5N02
5[ppm]    Multiplicity   -/(Hz) Integral
1.30	triplet	7	3
3.00	singlet	-	6
4.25	quartet	7	2
6.65	dublet	8	2
7.80	dublet	8	2
tori'. 26, 2SL7 O^Vd*
Task 2: CnHi5N02
5[ppm]    Multiplicity   J(Hz)    Integral ^       ff-^ O
1.30 b triplet 7 3 N
3.00 e singlet - 6
4.25 a quartet 7 2
6.65 d dublet 8 2
7.80 c dublet 8 2
Ondřej Jurček
26, 2SL7 O^Vd*
Task 3: Ci2Hi7NO - ^/COSY
!H NMR (300 MHz, CDCI3)
01    00    h OD
•í"   oi   o r-
(»1    Cl    Cl M
<j  in  oi  o  w h
Cl     rH     (C (\ O
oj   oj   h   -h   en cn
in m
m m m
oa • m in cn
inHPiHin«icri^co(ncn(ncninH
HO^COr-^mfOHCO^fOHCDCNO
u)inininir)inmin<noi(nonoicncn
•H  o  o ^
LLL L L L l J U J I J LAS
111111111111111111111111111111111111111111111111111111
7.7   7.5   7.3   7.1 ppm
1111111111111111111 > 1111111111111111111111111111111111111111111111111111111111111111111111111111 2.6   2.4   2.2   2.0   1.8   1.6   1.4   1.2 ppm
"~|-1-1-1-1-1-1-r
9 8
I 1 1 7
I 1 1 6
I 1 1 1 1 I 5 4
I 1 1 3
~l-[—1-r
2
—r
i
0.92 1.08
1.84 1.00
2.08 2.05 2.91
2.94 2.02
ppm
Ondřej Jurček
26, 2SL7 O^Vd*
Task 3: Ci2Hi7NO - ^/COSY
COSY (300 MHz, CDCI3)
Ondřej Jurček
F2
(ppm)
1-
3-
4
5
8
es &
9
26, 2&7 O^Vd*
Task 3: Ci2Hi7NO - ^-"C/HSQC.HMBC
HSQC (300 MHz, CDCI3)
Ondřej Jurček
F2
(ppm)
1-
I I  I  I--MM
MM--MM
MM--MM
7  T  T  1       I   1   1 ■
MM--MM
Mil--MM
i i i i    n i r
Mil--MM
140 120
100 80 60
Fl (ppm)
40
20
torí 26, 2B.7 •OTty(&>>
Task 3: Ci2Hi7NO - ^-"C/HSQC.HMBC
HMBC (300 MHz, CDCI3)
ifctMVNvitummii.mwirijiivi^i.imiiMi^ii^
F2
(ppm)
1-
2
3
4
5
O o
rl II I 1111   I I I I I I I I I   I 1 1 I | I I l l   l l l i II i l l   li li   l li l   l l l l 11 l l l   l l i i | l l l l   I I I I I I I I I   I 1 I I 1 I I 1 1   l l l l II l l l
180        160        140        120        100 80 60 40 20
Ondřej Jurček
26, 2B.7 •OTty(&>>
XH NMR (300 MHz, CDCI3)
l|l'"|""l""l""l""l""í""l""l""l""l |IIII|IIII|IIII|MII|IIII|IIII|IIII|IIII|IIII|IIII|IIII|IIII|IIII|IIII|MII|IIII|IIII|MII|IIII|
7.7   7.5   7.3   7.1     ppm 2.6   2.4   2.2   2.0   1.8   1.6   1.4   1.2 ppm
'fl
0.92 1.08
1.84 1.00
2.08 2.05 2.91
2.94 2.02
COSY(300 MHz, CDCI3)
F2
(ppm)
1-
i i i i—i i i i—i i i i—i i i i
1K Kl
f1111 f
9876543210 Fl (ppm)
Ondřej Jurček
m
26, 2SL7 O^Vd*
General comments
• inspect molecular formula CmHh0oNnXx: Degree of unsaturation m + 1 — 0.5(h + x — n)
• identify signals of CH3 and exchangeable protons in ID 1H sectrum
9 arbitrary numbering (e.g., from lower to higher value of chemical shift) of resolved resonances in all spectra
• identification of the individual spin systems using DQF-COSY
• resolve geminal protons using HSQC
• connect molecular fragments/isolated spins using HMBC, NOESY
specify the stereochemistry (relative configuration) by means of J— and NOE interaction
• in ID spectrum bottom blue numbers are integrals, labels in violet frames contains the arbitrary label (A-N), multiplet specification (use with caution, automatically determined), and position of a signal in ppm
9 UnHa-UnHb in 2D reffers to correlation of protons a and b of unknown compound Un
CO
73
ro
o
jo
-t co
ro — co
O
o
U) U)
ro
ro
□
4
.....
P
9
U) 4i
149.55 J>
■ 148.60 Q]
-138.75 O
-134.77 q
-123.50 m
77.26 77.00 76.75 68.84
- 56.98 Q
- 4034 X -35.19 —
- 22.59 c_
O D O
CO
CmHi.No in CDCľ
CD
LU
Q O
CG <
2     s 4-10"14"2
56 78
ppm
30 40 50 60 70 \- 80 90 t-100 110 120 -130 -140 150
(D
LU
Q O
00
<
3 4
56 78
" i"" i'"11" ■11" ■ ■ i1''11 ■ ■" i"" i ■1" i" ■1 r" ■ i"" i"" i""—
8.5  8.0  7.5  7.0 6.5  6.0  5.5  5.0 4.5  4.0  3.5  3.0 2.5  2.0 ppm
■ i1" 11 ■1111" " ľ1" i " " i ■1" i " ■1 r 1 ■ ■ i1 ■ ■ ■ i ■ ■ ■ i " 1111 ■ ■ i "111 ■ 8.5  8.0  7.5  7.0 6.5  6.0 5.5  5.0 4.5  4.0 3.5  3.0 2.5 2.0
ppm
30 40
\r 50
60 h 70 80 90 f-100 110 120 r 130 r 140 -150 ppm
"C^H HSQC
13C/H HMBC
CmHi.No in CDCľ
CD
LU
Q O
CG <
-10' '14^2
d      3 4
5 6 7d
I i 10,11,12
ppm
30 40 50 60 70 \- 80 90 t-100 110 120 -130 -140 150
(D
LU
Q O
00
<
3 4
5 6 78
" i"" i'"11" ■11" ■ ■ i1''11 ■ ■" i"" i ■1" i" ■1 r" ■ i"" i"" i""—
8.5  8.0 7.5  7.0 6.5  6.0  5.5 5.0 4.5 4.0 3.5  3.0 2.5  2.0 ppm
■ i1" 11 ■1" i " " ľ1" i " " i ■1" i " ■1 r 1 ■ ■ i1 ■ ■ ■ i ■ ■ ■ i " 1111 ■ ■ i "111 ■ 8.5  8.0  7.5  7.0 6.5  6.0 5.5  5.0 4.5  4.0 3.5  3.0 2.5 2.0
ppm
30 40
\r 50
60 h 70 80 90 f-100 110 120 r 130 r 140 -150 ppm
"C^H HSQC
13C/H HMBC
ID *H of CioH200
4.0
3.5
3.0
2.5
fl (ppm)
2.0
1.5
1.0
0.5
Jan Novotný
Q3V(3v
1H-13C HSQC and NOESY
UnC1-Ha UnC2-Hd ^
UnC3-Hqs^^
UnC4 _
UnC5-HE— j
UnC6-HľL UnC4-Hf
UnC7-Hj
\
Á
UnC7-Hb UnC8-Hk UnC8-Hc
\ 1 /
UnC9-Hg
UnC10-Hm
\
20
H
30
E
40   B 2
3
50 3
60
4^
70
4 2
C02-1H (ppm)
6
_i_
uhHa-Hn
4 —i—
_2
UnHd
UnHk-Hr
UnHd-Hk
UnHe-Hm llnHa Hm   Unŕla"HI   I UnHb-Hj , m   UnHa-Hm      i       f   . UnHb-^i
/
/ UnHf-Hj YV lnMÄ UnHg-H* \UnHf-Hi
UnHg-Hn UnHf-H«L-----uu Hm    J ^UjßHg-Hi
"UnHh-
UnHI-H
UnHk-Hm
UnHh-Hk£ří ©
UnHh-Hj . 4 #
UnHm-Hn
\
!
4
O 0
I
co2- 1H (ppm)
2
3
Task 1: J-con necti vity of C10H20O
5
J_I_L
J_I_L
J_I_I_L
J_I_I_L
2_
J_I_I_L
J_I_I_L
E
a. 2
Cl e~
UnHc-Hm
UnHg-Hm
UnHk-Hmi
UnHb-Hj UnHc-Hk     \ UnHb"
UnHb-Hh
UijiiHb-Hf
UnHf-HjTUnHg-Hi
UnHh-Hk^1
UnHh-Hj UnHj-ffk
v
UnHdl " ■'
UnHe-HV UnHf-Hi
I
r
h
3^
UnHm-Hn
Jan Novotný
1
i
1
□ S1
i =
Task 1: J-connectivity of C10H20O
• methyls la,2d connected to CH 51
O remaining crosspeak of CH 51 to CH 9g
• methyls la,2d diastereotopic =>- chiral carbon 9
E
a 2
4
_i_
UnHg-Hm
-1-
4
UnHb-Hj
UpHb-Hf
n—m\
^ \UnHe-Hti V
3 2 C02- 1H (ppm)
I
Jan Novotný
□
S1
4/5
Task 1: J-con necti vity of C10H20O
5
J_I_L
E
o. 2
Q.
3^
J_I_L
J_I_I_L
J_I_I_L
2_
J_I_I_L
J_I_I_L
UnHc-Hm
UnHg-Hm'
UnHk-Hmi
UnHm-Hn
Jan Novotný
UnHb-Hj UnHc-Hk     \ UnHb-Hij
UnHb-Hh
UijiiHb-Hf
T-V—í-r->-V£
UnHf-Hi
\
UnHf-Hb
I
f
h
1
□ S1
i =
Task 1: J-connectivity of C10H20O
• CH 9g has crosspeaks with deshielded 10m => OH group (n)
9 CH 9g has two crosspeaks with diastereotopic protons 4if
E
a 2
4
_i_
UnHm—Hn
UnHb-Hh UnHb-Hj . U*Hb-Hf
UnHb-Hil
fUnHf-
UnHf-Hi _® JO_0
S> iff
3 2 C02- 1H (ppm)
I
Jan Novotný
□
S1
4/5
Task 1: J-con necti vity of C10H20O
5 4 3 2 1
-I_I_I_I_I_I_I_I_I_I_I_I_I_I_I_I_I_I_I_I_L
Task 1: J-connectivity of C10H20O
• CH lOm connected with
CH2 8ck 5 4 3 2 1
5 4 3 2 1
C02- 1H (ppm)
Jan Novotný
□
4/5
Task 1: J-con necti vity of C10H20O
Jan Novotný
Task 1: J-connectivity of C10H20O
CH2 8ck weakly coupled with CH2 7bj =>- closing ring
protons b and k coupled to CH 6h which is connected to methyl 3e
other expected crosspeaks in DQ-COSY c rowd ed / ove r I a p p ed, found topology confirmed
in HMBC (3/4Jhc) a,, A  H 2d
hUC
10.
CH,
8,
H'
H
5	
^9	b
H1	
	
	H
E
& 2 X
3
4
_i_
UnHc-Hm
DnHg-Hm"
UnHJc-Hm™
UnHm—Hn
—i—
4
DnHa-Hl OnHd-Hl
DnHg-Hl
-
UnHB-HJ i UpHb-Hf
UnHc-Hk    \ OnHb-Hif I»
^^^V , \   \ J Ivbsi-bJ',/
UnHf-
3 2 Cö2- 1H (ppm)
I
Jan Novotný
□
4/5
Task 1: J-connectivity of C10H20O
3(H
35 A
E
Q. O.
O
co
40H
45H
5(H
Jan Novotný
_i_i_i_
j_i_i_
HMBC
«nC6-Hin^\ W6-Hb UnC6-Hk—----s T
UnC7-He^ UnC6-Hf
h 30
UnC7-Hk
h 35
UnC7-Hj
UnC7-Hb
UnC7-Hi
h 40
UnC8-He
UnC8-Hm
UnC8-Hn
UnC8-Hk    „ft „. \ UnC8-Hb UnC8-Hh
\
^u*iv*o—au
h 45
UnC9-Hn UnC9-Hm
\ \
UnC9:H1    UnC9-Hd ""^-f \  UnC9-Hi \ Un""Ha
UnC9-Hg
h 50
=
Task 1: J-connectivity of C10H20O
• CH2 8ck weakly coupled with CH2 7bj => closing ring
• protons b and k coupled to CH 6h which is connected to methyl 3e
9 other expected crosspeaks in DQ-COSY c rowd ed / ove r I a p p ed, found topology confirmed
in HMBC {3/4JHc)
a      1  H 2 d H3a N XH
MENTHOL
Jan Novotný
30-
35
CL Q.
O
co
40
45
50
4
_1_
UnC6-Hh
UnC6-He
HMBC
UnC6-Hi UnC6-Hk
V
UnC6-Hb - 30
UnC7-He^ UnC6-Hf
UnC7-Hk'
UnC7-H] \
\
- 35
UnC7-Hb
UnC7-Hi
UnC8-He
UnC8-Hm   UnC8_Hk \ UnC8-Hb
UnC8-Hn \ xUnC8-Hh \ /
\ \ \\y
nnrQ-Hl UnC8-Hc UnC9-H„ UnC.-H, U„C9-Ha
\ \ \     -   \ ^/
UnC9-Hg
2
40
45
■ 50
Q)2-1H (ppm)
□ fl1
□
S1
4/5
Task 1:
Stereochemistry of menthol C10H20O
lo2:
homonuclear/heteronuclear couplings
• large couplings preserved in ID slices of HSQC:
axial H - 2 visible interactions (geminal and vicinal) x equatorial H - only geminal a ID TOCSY: selective decoupling =^ simplification of complex multiplets
• DQF-COSY: analysis of phase sensitive spectrum
o lo3: NOE contacts (axial strong)
Hx
HB 3JX.A = small 3JX.B = large
C
3JHC= 6-8 Hz 2JHr = 5-7 HZ
HB
3JX.A = small 3JX.B = small
C
=  1-3 HZ
R
2JHr = 0-2 Hz
Jan Novotný
□
5/5
Task 1:
Stereochemistry of menthol C10H20O
lo2:
homonuclear/heteronuclear couplings
• large couplings preserved in ID slices of HSQC:
axial H - 2 visible interactions (geminal and vicinal) x equatorial H - only geminal a ID TOCSY: selective decoupling =^ simplification of complex multiplets
• DQF-COSY: analysis of phase sensitive spectrum
o lo3: NOE contacts (axial strong)
Jan Novotný
□
5/5