Nuclear Magnetic Resonance

Spectroscopy

Hugh Benhardt
February 29, 2008
Outline

  Nuclear Magnetic Resonance (NMR) theory

  Introduction to NMR spectroscopy
  Methods of NMR spectroscopy
  Modern applications
Intrinsic Properties of Nuclei

  Spin: rotation of a body around its own center of mass

  Nuclei with an odd number of protons/neutrons have spin
(e.g., 1H, 13C, 19F, 31P)
  As positively charged particles, these nuclei have an intrinsic
magnetic moment
  In an magnetic environment, nuclei behave like magnets
Orientation of Spinning Nuclei

  No external magnetic field: nuclear spins orient randomly

  External magnetic field: spinning nuclei align parallel or antiparallel
  Parallel orientation: lower energy, more favorable
Nuclear Magnetic Resonance

  When oriented nuclei are subjected to electromagnetic radiation at

the proper frequency
–  Energy absorption occurs
–  Lower energy state nuclei “spin-flip” to the higher energy state
–  Magnetic nuclei in resonance with applied radiation
  Resonant frequency dependent on strength of magnetic field and
identity of the nucleus
Shielding

  All 13C and 1H nuclei do not

resonate at the same frequency
  Each nucleus has its own local
magnetic field that opposes the
applied magnetic field
  Beffective = Bexternal – Blocal
  Nuclei in different electronic
environments resonate according to
their effective fields
Outline

  NMR theory
  Introduction to NMR spectroscopy
  Methods of NMR spectroscopy
  Modern applications
Purpose of NMR Spectroscopy

  Determine structural information

  Identify type of polymer
  Observe the effects of a reaction
mechanism
  Map the C-H framework of a
chemical compound
Experimental Setup

  Polymer sample dissolved in solvent

(e.g., cdcl3)
  Sample placed in a thin glass tube
  Tube inserted into an NMR spectrometer
between the poles of a magnet
  A magnetic field causes nuclei to align
  Nuclei irradiated at constant frequency
and varied magnetic field strength
  Each nucleus resonates at a different
strength and is displayed as a separate
peak on an NMR spectrum
NMR Spectrum

  Horizontal axis: effective field strength

  Vertical axis: intensity of energy absorption
Chemical Shift

  Chemical shift (δ): position on chart where a nucleus

absorbs energy
  δ = (observed chemical shift)/(spectrometer frequency)
  1 δ = 1 ppm (parts per million)
–  1 millionth of 300,000,000 Hz: 1 ppm = 300 Hz
  Relative term to the spectrometer frequency
  Allows comparison of results from different instruments
of varied strength
Chemically Equivalent Environments

  Absorption peak: reflective of electronic environment

  Nuclei in chemically equivalent environments show single
absorption peak
Absorption Peaks

  The area under each peak is proportional to the number of particles

  Integration of each peak yields the number of atoms at each
chemical shift
  “Stair-step” line
–  Ratio of step heights is proportional to the number of particles in
each peak
Outline

  NMR theory
  Introduction to NMR spectroscopy
  Methods of NMR spectroscopy
–  Carbon13 NMR
–  Hydrogen1 (Proton1) NMR
–  High frequency NMR
–  Solid-state NMR
  Modern applications
Carbon13 NMR

  Carbon13
–  Not commonly found in nature
–  Only carbon with nuclear spin
  Weak signals produce “noisy” results
  Signal averaging
–  Eliminates background noise
–  Increases time of procedure
  FT-NMR
–  Pulse of energy covers range of frequency at a constant
magnetic field strength
–  Records signals simultaneously
–  Fourier transforms manipulate data to produce normal
spectrum
Chemical Shift of Carbon Atoms

  Chemical shift is affected by the electronegativity of nearby atoms

  Electronegative atoms draw electrons away from carbon atoms,
leaving them deshielded
  Carbons bonded to oxygen, nitrogen, or halogen are located
downfield of alkane carbons
  Chemical shifts range from 0 - 220 δ
Uses of Carbon13 NMR

  Determine structural information

–  Number of nonequivalent carbon atoms in a molecule
–  Electronic environment: number of hydrogen atoms
attached to each carbon
Carbon13 vs. Hydrogen1 NMR

  Carbon and hydrogen atoms require a different amount of

energy to “spin-flip”
  Spectrometers of different field strengths required to resonate
each nucleus type
–  1H: 0 – 10 δ downfield from TMS
–  13C: 0 – 220 δ downfield from TMS
  Carbon and hydrogen atoms must be observed separately
Outline

  NMR theory
  Introduction to NMR spectroscopy
  Methods of NMR spectroscopy
–  Carbon13 NMR
–  Hydrogen1 (Proton1) NMR
–  High frequency NMR
–  Solid-state NMR
  Modern applications
Hydrogen1 NMR

  The terms Hydrogen1 NMR and Proton1 NMR may be used

interchangeably
  Similar to 13C NMR, 1H NMR determines the number of
hydrogen atoms in a molecule
  Chemical shifts range from 0 – 10 δ
Spin-Spin Splitting
  The magnetic field of one nucleus may affect nearby hydrogen atoms
  Coupling: interaction of spins between adjacent hydrogen atoms
causes multiple absorption peaks
Spin-Spin Splitting (continued)

  Each hydrogen nucleus may align with or against a neighboring

magnetic field
  Nuclear realignment shifts the effective field of the neighboring
nucleus, changing the applied field necessary to cause resonance
  Coupling constant (J): distance between peaks
General Rules for Spin-Spin Splitting

  Chemically equivalent protons do not show spin-spin splitting

  Protons with n equivalent neighboring protons are split into a
multiplet of n+1 peaks with coupling constant J
  Two groups of protons coupled to each other have the same
coupling constant J
  Protons farther than 2 carbons apart do not couple
  Carbon spin-spin splitting:
–  13C atom are rarely adjacent in nature
–  Spin-spin splitting does not occur
Uses of Hydrogen1 NMR

  Determine structural information

–  Number of nonequivalent hydrogen atoms in a molecule
–  Electronic environment of each atom
–  Identify product of chemical reactions
Outline

  NMR theory
  Introduction to NMR spectroscopy
  Methods of NMR spectroscopy
–  Carbon13 NMR
–  Hydrogen1 (Proton1) NMR
–  High frequency NMR
–  Solid-state NMR
  Modern applications
High Frequency NMR

  Complex chemical
compounds have more
overlap of absorption
peaks due to multiple
electronic environments
  Increasing field strength
produces a much larger
spectrum
  Absorption peaks are
more spread out
  Easier for interpretation
of results
Outline

  NMR theory
  Introduction to NMR spectroscopy
  Methods of NMR spectroscopy
–  Carbon13 NMR
–  Hydrogen1 (Proton1) NMR
–  High frequency NMR
–  Solid-state NMR
  Modern applications
Solid-State NMR (SSNMR)

  Solids (crystals, gels, etc.) are dominated by anisotropic

(direction-dependent) interactions
  Anisotropic interactions have substantial influence on
the behavior of nuclear spins in solids
  Anisotropic interactions:
–  Chemical shift anisotropy
–  Internuclear dipolar coupling
Solid-State vs. Solution-State NMR

  Solution-state NMR
–  Solution: signal averaging compensates for anisotropic
interactions
–  Solids:
  Molecules held still; no signal averaging
  Interactions broaden spectral lines: less resolution
  Solid-state NMR
–  Magic Angle Spinning (MAS): spinning sample at the magic
angle with respect to the direction of the magnetic field
–  Decoupling of nuclear spin interactions
–  Increases resolution of NMR spectra
Outline

  NMR theory
  Introduction to NMR spectroscopy
  Methods of NMR spectroscopy
  Modern applications
–  Chemistry
–  Biology
–  Medicine
Chemistry

  Determine molecular structure of

different chemical compounds,
including polymers
  Identify products of chemical
reactions
  Means to study dynamics, kinetics,
and thermodynamics of chemical
systems
Biology

  Protein NMR
–  Obtain structural information on
complex proteins
–  Multidimensional experiments
needed to correlate frequencies
with distinct nuclei
Medicine
  Magnetic Resonance Imaging (MRI)
–  Insight into the structure of the
human body
–  Sharp contrast in soft tissue
  Clinical applications
–  Cardiovascular system
–  Musculoskeletal system
–  Oncology
–  Neurology
Summary

  Nuclear Magnetic Resonance (NMR): change in alignment of

atomic nuclei from preferred energy states due to intrinsic
magnetic properties
  Used to determine structural framework of chemical
compounds
  Applicable for multiple disciplines
–  Medicine
–  Chemistry
–  Biology
Questions?