Basics in NMR

SPECTROSCOPIC METHODS
OF ORGANIC MOLECULES
Professor Ioannis P. Gerothanassis

Department of Chemistry, University of Ioannina
FEBRUARY 2018
SPECTROSCOPIC METHODS
OF ORGANIC COMPOUNDS
Concepts and Applications of NMR
Professor Ioannis P. Gerothanassis

Department of Chemistry, University of Ioannina
3
What is NMR ?
• A spectroscopic technique that gives information about

the number and types of atoms in a molecule.
• Nuclear magnetic resonance spectroscopy is a powerful
analytical technique used to characterize molecules by
identifying e.g. hydrogen-carbon frameworks within
molecules.
Nuclear Magnetic Resonance
In the Nucleus Involves At the Nucleus

Magnets
http://www.authorstream.com/Presentation/Darshantelange-1542377-nmr-spectroscopy-ppt/
4
Nuclear Magnetic
Nuclear Spin
• A nucleus with an odd atomic number or an odd
mass number has a nuclear spin.
• The spinning charged nucleus generates a
magnetic field e.g. acts like a bar magnets.
5
Precessional Motion:
Consider a spinning top, in which the
spinning axis of the top moves slowly
around the vertical. This is precessional
motion and the top is said to be precessing
around the vertical axis of earth’s
gravitational field.
Precessional Frequency:
Likewise the proton (tiny magnet)
precesses about the axis of the external
magnetic field in the same manner as
above.
6
The spinning proton in an external magnetic field

The nuclear magnetic resonance process; absorption occurs
when ν =ω
ΒΟ
7
Two Energy States in Quantum Chemical

Terms
 The magnetic fields of
the spinning nuclei will
align either with the
external field, or
against the field.
 An r.f. with the right
amount of energy can
be absorbed and
cause the spinning
proton to flip.
8
E, Magnet Strength and 

The energy difference ΔΕ is proportional to the
magnetic field strength B0 and .
E = h =  h B0
2
• Gyromagnetic ratio, , is a constant for each nucleus

(26,753 s-1gauss-1 for 1H).
9
Rf Field vs. Magnetic Field for a Proton

E
60 MHz
500 MHz
E = h/2(Bo)
100 MHz
1.41 T 2.35 T 11.74 T
Bo
10
Rf /  values for Some Nuclei

Nuclei Rf (MHz) Bo (T) /2 (MHz/T)
1H 500.00 11.74 42.58
13C 125.74 11.74 10.71
2H 76.78 11.74 6.54
19F 470.54 11.74 40.08

31P 202.51 11.74 17.25
11
Where Nuclei Resonate at 11.74T
12 ppm
2H
RCO2H 12 ppm
1H (TMS)
~380 ppm
31P (H3PO4) ~220 ppm

CH4
76.78 220 ppm 19F (CFCl3)
13C
500.00
PX3 CPH2
-SO2F -CF2-CF2-
202.51 470.54
125.74
R2C=O RCH3
100 200 300 400 500 MHz
http://textlab.io/doc/9993536/the-basics-of-nmr--powerpoint-
12
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Magnetic Shielding
• If all protons absorbed the same amount of
energy in a given magnetic field, not much
information could be obtained.
• But protons are surrounded by electrons that
shield them from the external field.
• Circulating electrons create an induced
magnetic field that opposes the external
magnetic field.
Beff=Bexternal-Bshielding
http://wps.prenhall.com/wps/media/objects/340/348272/Instructor_Resources/Chapter_13/Wade13.ppt
14
• Such shifting in position of NMR absorption signals which

arise due to the shielding or deshielding of proton by
surrounding electrons is called the Chemical shift.
15
Shielding or Deshielding Protons in a Molecule

• Depending on electronic environment protons in
molecules are shielded or deshielded by different
amounts.
16
• Shielded proton shows absorption of signals to right side and

deshielded protons at left side of spectrum.
17
NMR Signals
• The number of signals shows how many different
kinds of protons are present.
• The location of the signals shows how shielded or

deshielded the proton is.
18
The NMR Graph
Integral: 1 3
19
NMR Signals

• The intensity of the signal shows the number of

protons of that type.
20
IR spectrum of methanol
21
• Any proton or set of protons which absorb at higher frequency

than TMS is given a positive value for (δ).
• The value of δ for a substance relative to TMS can be obtained

by Chemical shift, ppm.
Shift high frequency from TMS (in Hz)

δ=
Spectrometer frequency (in MHz)
• The value of chemical shift, δ expressed in ppm and their value

is between 0 to 10 in the δ scale.
22
Delta Scale
23
Hydrogen Chemical Shifts
ppm
http://rd.nctu.edu.tw/web.case/nctu-rd-2/upload/ckeditor/20150410151405.pdf
2424
Classification of the Chemical Shift of the -OH Functional Groups
Kontogianni et al., Org. Biomol. Chem, 2013, 11, 2013-2025

Siskos et al., Org. Biomol. Chem, 2013, 11, 7400-74011
25
2
3
O
H
O
4
17.5-20 ppm
14.75 ppm 14.17 ppm

A C
5 4
O O
H
12-13 ppm
26
Effect of Strength of Hydrogen Bond on 1H Chemical Shift
27
http://www.authorstream.com/Presentation/knowledgexplorers-2245188-anwar-nmr/
28
most
deshielded
29
30
Hydrogen Chemical Shifts
ppm
31
32
Aromatic Protons, 7-8 ppm
The aromatic protons

are deshielded!
33
Aldehyde Proton, 9-10 ppm
Electronegative
oxygen atom
34
Vinyl Protons, 5-6 ppm
35
Acetylenic Protons, 2.5 ppm
36
NMR Signals

• The intensity of the signal shows the number of

protons of that type.
• Signal splitting shows the number of protons on

adjacent atoms.
37
Spin-Spin Splitting
1,1,2-Tribromoethane
Nonequivalent protons on adjacent carbons.
38
Doublet: 1 Adjacent Proton
39
Triplet: 2 Adjacent Protons
40
Spin-Spin Coupling Constants

• Distance between the peaks of multiplet.
• Measured in Hz.
• Not dependent on strength of the external field.
• Multiplets with the same coupling constants may

come from adjacent groups of protons that split
each other.
• Protons separated by four or more bonds will ?
not couple.
41
Values for
Coupling Constants
42
Splitting Tree
a c
H H
C C
Hb
http:///
43
44
https://translate.google.gr/translate?hl=el&sl=en&u=http://bionmr.unl.edu/courses/chem99
1a_-_intro_to_nmr/lectures/chapter-3-NMR-coupling.ppt&prev=search
45
46
Hydrogen and Carbon
Chemical Shifts
47
Carbon-13
• 12C has no magnetic spin.
• 13C has a magnetic spin, but is only 1% of the carbon
in a sample.
• The gyromagnetic ratio of 13C is one-fourth of that
of 1H.
• Signals are weak, getting lost in noise.
• Hundreds of spectra are taken, averaged.
48
The NMR Spectrometer
continuous change in frequency
49
50
Periodic variation of experimental nuclear

shielding ranges
51
Fourier Transform NMR

• Nuclei in a magnetic field are given a radio-frequency pulse
close to their resonance frequency
• The nuclei absorb energy and process (spin) like little tops
• A complex time domain signal is produced, then decays as

the nuclei lose energy
• Free induction decay is converted to spectrum
http://www.chemistry-solutions.com/wp-content/uploads/2015/07/CHE-242-Unit-5-Chapter-13.ppt
52
excitation pulse (μs)

53
http://rutchem.rutgers.edu/~nmurali/nmr_course/Chem_542_Spring2010_Lecture_3.pdf
54
1H NMR spectra of caffeine 13C 13C

NMR spectra of caffeine NMR spectra of caffeine
8 scans ~12 secs 8 scans ~12 secs 10,000 scans ~4.2 hours
55
56
Spin population NMR - Sensitivity

57
Nuclear Spins in Bo
• In an applied field strength of 7.05T the difference
in energy between nuclear spin states for
– 1H is approximately 0.120 J (0.0286 cal)/mol, which
corresponds to a frequency of 300 MHz (300,000,000
Hz).
– The population difference of the two nuclear spin
states is only ∼10-5 and, therefore, has meaning only
at the Avogadro number!
http://www.authorstream.com/Presentation/kesarwaniarti-1755809-nmr-spectroscopy/
58
NMR Sensitivity
• Relative sensitivity of 1H, 13C, 15N and other nuclei NMR spectra depend on
 Gyromagnetic ratio ()
 Natural abundance of the isotope
-  intrinsic property of nucleus can not be changed
  C)3 for 13C is 64x   N)3 for 15N is 1000x
1H is ~ 64x as sensitive as 13C and 1000x as sensitive as 15N !
Consider that the natural abundance of 13C is 1.1% and 15N is 0.37%
relative sensitivity increases to ~6,400x and ~2.7x105x !!
59
http://www.authorstream.com/Presentation/solairajan87-1493928-nmr-spectroscopy/
60
N.O.E. Transient NOE and Distances

61
N.O.E. Example “steady-state”
Ha Hb
Prof. Claudio Santi Dipartment of Chemistry and Technology of Drugs University of Perugia
62
NMR and Chemical Exchange

63
What information can NMR spectroscopy provide
Structural Theory NMR equivalent

Qualitative composition Choice of nucleus / isotope
Quantitative composition Signal intensities / integration
Functional groups Chemical shifts
Connectivity J (scalar) couplings
Spatial relationship of atoms NOE (distance)

J couplings (dihedral angle)
Dynamics Chemical exchange, lineshape

broadening
64
Books in NMR - Web Sites

Solving Problems with NMR Spectroscopy by Atta-ur-Rahman and Muhammad Iqbal Choudhary
http://www.elsevier.com/books/modern-nmr-techniques-for-chemistry-research/derome/978-0-08-
032513-2
Modern NMR Techniques for Chemistry Research by A.E. Derome, University of Oxford, UK
http://www.elsevier.com/books/modern-nmr-techniques-for-chemistry-research/derome/978-0-08-
032513-2
Modern NMR spectroscopy, a guide for chemists, by J. K. M. Sanders and B. K. Hunter

http://onlinelibrary.wiley.com/doi/10.1002/mrc.1260251217/abstract
Nuclear Magnetic Resonance : Volume 3 by R K Harris

http://pubs.rsc.org/en/Content/eBook/978-0-85186-272-9#!divbookcontent
NMR Spectroscopy: Basic Principles, Concepts and Applications in Chemistry, by H. Gunther

http://www.wiley-vch.de/publish/en/books/ISBN978-3-527-33000-3/
Carbon 13 NMR Spectroscopy, by Hans-Otto Kalinowski, Stefan Berger and Siegmar Braun
http://eu.wiley.com/WileyCDA/WileyTitle/productCd-0471913065.html
65
Some NMR Web Sites

The Basics of NMR by J.P. Hornak Hypertext based NMR course
http://www.cis.rit.edu/htbooks/nmr/nmr-main.htm
Integrated Spectral Data Base System for Organic Compounds

http://www.aist.go.jp/RIODB/SDBS/menu-e.html
Educational NMR Software All kinds of NMR software

http://www.york.ac.uk/depts/chem/services/nmr/edusoft.html
NMR Knowledge Base A lot of useful NMR links

http://www.spectroscopynow.com/
NMR Information Server News, Links, Conferences, Jobs

http://www.spincore.com/nmrinfo/
Technical Tidbits Useful source for the art of shimming

http://www.acornnmr.com/nmr_topics.htm
BMRB (BioMagResBank) Database of NMR resonance assignments

http://www.bmrb.wisc.edu/
66
STRUCTURE EUCIDATION USING NMR

SPECTROSCOPY – AN EXAMPLE
THE CASE OF THE FLAVONOID QUERCETIN

67
68
69
70
2.2 Hz 8.5Hz, 2.2 Hz 2.02 Hz 2.02 Hz
71
TWO DIMENSIONAL SPECTRA

THE 1H-1H COSY EXAMPLE
Jacobsen, NMR Spectroscopy Explained, Wiley, 2007

72
Homonuclear 2 Dimensional Experiment:
1H – 1H COSY (COrrelated SpectroscopY)
1H
1H
73
Homonuclear 2 Dimensional Experiment:
1H – 1H TOCSY (TOtal Correlated SpectroscopY)
1H
1H
Heteronuclear 2 Dimensional Experiment: 74
1H – 13C HSQC (Heteronuclear Single Quantum Spectroscopy)
13C
1H
75
Heteronuclear 2 Dimensional Experiment:

1H – 13C HMBC (Heteronuclear Multiple Bond Correlation)
H5’ H8
H6
H7’ H6’
2’
8 1
B
5’ 13C
A C 6’
6 4
C-OH
C-OH
C=O
1H

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SPECTROSCOPIC METHODS

Professor Ioannis P. Gerothanassis

Concepts and Applications of NMR

Professor Ioannis P. Gerothanassis

• A spectroscopic technique that gives information about

In the Nucleus Involves At the Nucleus

The spinning proton in an external magnetic field

Two Energy States in Quantum Chemical

E, Magnet Strength and 

• Gyromagnetic ratio, , is a constant for each nucleus

Rf Field vs. Magnetic Field for a Proton

1.41 T 2.35 T 11.74 T

Rf /  values for Some Nuclei

1H 500.00 11.74 42.58

13C 125.74 11.74 10.71

2H 76.78 11.74 6.54

19F 470.54 11.74 40.08

Where Nuclei Resonate at 11.74T

31P (H3PO4) ~220 ppm

100 200 300 400 500 MHz

• Such shifting in position of NMR absorption signals which

Shielding or Deshielding Protons in a Molecule

• Shielded proton shows absorption of signals to right side and

• The location of the signals shows how shielded or

The NMR Graph

• The location of the signals shows how shielded or

• The intensity of the signal shows the number of

• Any proton or set of protons which absorb at higher frequency

• The value of δ for a substance relative to TMS can be obtained

Shift high frequency from TMS (in Hz)

• The value of chemical shift, δ expressed in ppm and their value

Hydrogen Chemical Shifts

Kontogianni et al., Org. Biomol. Chem, 2013, 11, 2013-2025

14.75 ppm 14.17 ppm

Hydrogen Chemical Shifts

Aromatic Protons, 7-8 ppm

The aromatic protons

Aldehyde Proton, 9-10 ppm

Vinyl Protons, 5-6 ppm

Acetylenic Protons, 2.5 ppm

• The location of the signals shows how shielded or

• The intensity of the signal shows the number of

• Signal splitting shows the number of protons on

Doublet: 1 Adjacent Proton

Triplet: 2 Adjacent Protons

Spin-Spin Coupling Constants

• Multiplets with the same coupling constants may

The NMR Spectrometer

continuous change in frequency

Periodic variation of experimental nuclear

Fourier Transform NMR

• A complex time domain signal is produced, then decays as

• Free induction decay is converted to spectrum

excitation pulse (μs)

1H NMR spectra of caffeine 13C 13C

Spin population NMR - Sensitivity

-  intrinsic property of nucleus can not be changed

1H is ~ 64x as sensitive as 13C and 1000x as sensitive as 15N !

N.O.E. Transient NOE and Distances

N.O.E. Example “steady-state”

NMR and Chemical Exchange

What information can NMR spectroscopy provide