Module III
MAGNETIC RESONANCE SPECTROSCOPY
Nuclear Magnetic Resonance (NMR) spectroscopy is a powerful analytical technique used to
determine the structure of organic compounds, understand molecular dynamics, and study complex
mixtures.
• It identifies the carbon-hydrogen framework of an organic compound. Using this method and
other instrumental methods including infrared and mass spectrometry, scientists are able to
determine the entire structure of a molecule.
• Even though there are many other spectrometers including C-NMR and N-NMR, hydrogen (H-
NMR) was the first and is the most common atom used in nuclear magnetic resonance
spectroscopy.
• Many nuclei may be studied by NMR techniques, but hydrogen and carbon are most
commonly available. NMR gives information about the number of magnetically distinct atoms
of the type being studied.
• When hydrogen nuclei (protons) are studied, for instance, one can determine the number of
each of the distinct types of hydrogen nuclei as well as obtain information regarding the
nature of the immediate environment of each type. Similar information can be determined
for the carbon nuclei.
Principle of Nuclear Magnetic Resonance (NMR) Spectroscopy
The principle behind NMR is that many nuclei have spin and all nuclei are electrically charged.
If an external magnetic field is applied, an energy transfer is possible between the base energy to a
higher energy level (generally a single energy gap).
The energy transfer takes place at a wavelength that corresponds to radio frequencies and when the
spin returns to its base level, energy is emitted at the same frequency.
The signal that matches this transfer is measured in many ways and processed in order to yield an
NMR spectrum for the nucleus concerned.
Dr. R. C. Kulkarni | KLS GIT, BGM
Module III
1. Magnet
Superconducting Magnet: NMR instruments typically use a powerful superconducting magnet to
generate a strong and stable magnetic field. The strength of this magnetic field is measured in teslas
(T), with common laboratory NMR spectrometers operating at 1.5 T, 7 T, or higher.
Field Homogeneity: The homogeneity of the magnetic field is crucial for obtaining high-resolution
spectra. Shims are used to fine-tune the field uniformity.
Dr. R. C. Kulkarni | KLS GIT, BGM
Module III
2. Radiofrequency Transmitter
RF Coil: The RF coil generates the radiofrequency pulses required to excite the nuclei. The frequency
of the RF pulses must match the Larmor frequency (the precession frequency of the nuclei in the
magnetic field).
Pulse Programmer: The pulse programmer controls the timing, duration, and sequence of RF pulses,
which can be varied to perform different types of NMR experiments.
3. Sample Probe
Sample Tube: The sample is usually dissolved in a deuterated solvent (to avoid interference from
the solvent's hydrogen atoms) and placed in a thin-walled glass tube.
Probe: The sample tube is inserted into the probe, which is placed at the center of the magnetic
field. The probe contains the RF coil that transmits and detects the NMR signals.
4. Receiver and Detector
RF Receiver: After the nuclei absorb the RF radiation and re-emit it during relaxation, the same coil
or a different coil detects these signals.
Amplification and Conversion: The weak NMR signals are amplified and converted from analog to
digital form for processing.
Fourier Transform: The digital signal is processed using Fourier transform to convert the time-
domain signal into a frequency-domain spectrum, which is what is typically analyzed.
5. Computer System
Data Acquisition and Processing: The computer system controls the instrument, collects data, and
processes the NMR spectra. Software is used for spectrum analysis, including peak picking,
integration, and interpretation of chemical shifts, coupling constants, and other parameters.
Types of NMR Spectroscopy
1
H NMR (proton NMR)
The 1H (also known as proton) NMR spectroscopy is limited to ordinary hydrogens or protons. It is
the most often used in laboratories. Because most organic compounds contain numerous hydrogen
atoms, and the hydrogen atoms absorb energy of different wavelengths depending on their bonding
environment, proton NMR spectra provide a wealth of information about molecular structure.
It gives the information:
• the number of distinct hydrogens in the compound
• Unique hydrogen ratios
• Each hydrogen’s chemical surroundings
• Hydrogens in close proximity
Carbon-13 magnetic resonance spectroscopy
Dr. R. C. Kulkarni | KLS GIT, BGM
Module III
C-13 NMR spectroscopy is used to analyze the carbons in a molecule. Although it is less prevalent
than H-1, it is nevertheless utilized in organic chemistry laboratories. Natural carbon is almost
entirely constituted of the carbon-12 isotope, which has no magnetic moment and hence cannot be
detected by NMR techniques. Carbon-13 (13C) atoms, which account for around 1% of all carbon
atoms, do, however, absorb radio-frequency waves in a way comparable to hydrogen. As a result,
13
C NMR is feasible, and the technique yields useful information on the structure of the carbon
skeleton in organic compounds. Because just one out of every hundred carbon atoms in a molecule
contains a 13C isotope, and 13C atoms absorb electromagnetic radiation very poorly, 13C NMR
signals are approximately 6,000 times weaker than proton signals.
C-13 NMR can provide the following data:
• The total number of distinct carbons in the compound
• The compound’s chemical environment
Nuclear Magnetic Resonance (NMR) spectroscopy is a powerful analytical technique with a wide
range of applications across various fields. Here are some of the key applications:
1. Structural Determination of Organic Compounds
• Molecular Structure Elucidation: NMR spectroscopy is widely used to determine the
structure of organic molecules, including identifying functional groups, carbon-hydrogen
frameworks, and stereochemistry.
• Conformational Analysis: NMR can provide information about the conformations (3D
arrangements) of molecules, which is crucial in understanding their function and reactivity.
2. Biological and Biochemical Applications
• Protein Structure and Dynamics: NMR spectroscopy is used to determine the three-
dimensional structures of proteins and nucleic acids in solution. It is also used to study
protein-ligand interactions, folding processes, and molecular dynamics.
• Metabolomics: NMR is employed in metabolomics to analyze and quantify metabolites in
biological samples, providing insights into metabolic pathways and disease states.
3. Material Science
• Characterization of Polymers: NMR spectroscopy is used to study the structure,
composition, and dynamics of polymers. It can provide information on the sequence of
monomer units, branching, and molecular weight distribution.
• Solid-State NMR: This technique is used to study materials that are difficult to analyze in
solution, such as polymers, crystalline materials, and complex mixtures. It helps in
understanding the atomic-level structure of solids.
4. Pharmaceuticals and Drug Development
• Drug Discovery and Development: NMR is used to analyze the structure of drug candidates
and their interactions with biological targets. It also aids in the identification of impurities
and degradation products in pharmaceutical formulations.
• Quantitative NMR (qNMR): Used for the accurate quantification of drug substances,
excipients, and impurities, providing a robust method for purity assessment.
Dr. R. C. Kulkarni | KLS GIT, BGM
Module III
5. Chemical Analysis
• Complex Mixture Analysis: NMR spectroscopy can analyze complex mixtures, such as those
found in natural products, without the need for separation techniques like chromatography.
• Reaction Monitoring: NMR is used to monitor chemical reactions in real-time, providing
insights into reaction mechanisms, intermediates, and kinetics.
6. Environmental Science
• Pollutant Analysis: NMR is used to identify and quantify pollutants in environmental
samples, such as soil, water, and air.
7. Food Science
• Food Authentication: NMR is used to authenticate food products, identify adulterants, and
study the composition of complex food matrices.
• Quality Control: It helps in monitoring the quality and safety of food products by analyzing
their chemical composition and detecting contaminants.
8. Forensic Science
• Drug Analysis: NMR is used in forensic laboratories to identify and quantify drugs, poisons,
and other substances in biological samples.
• Document Examination: NMR can analyze inks and papers in questioned documents,
helping in forgery detection.
DMSO-d₆ and TMS are commonly used substances in NMR spectroscopy for specific purposes, such
as acting as solvents or references.
DMSO-d₆ (Deuterated Dimethyl Sulfoxide)
1. Chemical Structure and Composition:
1. DMSO-d₆ is a deuterated form of dimethyl sulfoxide (DMSO), where six hydrogen atoms
in the DMSO molecule are replaced by deuterium atoms
2. Chemical Formula: (CD3)2SO
3. Use in NMR Spectroscopy:
Solvent: DMSO-d₆ is commonly used as a solvent in NMR spectroscopy, especially for
polar and high-molecular-weight compounds that are not soluble in less polar solvents
like chloroform-d (CDCl₃).
Deuterated Solvent: The presence of deuterium instead of hydrogen minimizes
interference in the 1^{1}1H NMR spectrum because deuterium produces a much weaker
NMR signal due to its lower gyromagnetic ratio and different nuclear spin (I=1).
Residual Solvent Peak: DMSO-d₆ has a residual 1H signal (due to trace amounts of non-
deuterated DMSO) that typically appears at about 2.50 ppm in 1H NMR spectra, which is
useful as an internal reference for chemical shifts.
4. Solvent Properties:
Dr. R. C. Kulkarni | KLS GIT, BGM
Module III
High Boiling Point: DMSO-d₆ has a high boiling point (189°C), making it suitable for studying
compounds at elevated temperatures.
Polar Aprotic Nature: Its polar nature allows it to dissolve a wide range of substances, including
ionic compounds, making it versatile for NMR studies.
TMS (Tetramethylsilane)
Chemical Structure: TMS is an organosilicon compound with the formula . It consists of a
silicon atom bonded to four methyl groups.
Chemical Formula: (CH3)4Si
Use in NMR Spectroscopy:
• Chemical Shift Reference: TMS is the standard reference compound in NMR spectroscopy
for both 1H and 13C NMR. By convention, the chemical shift of TMS is set to 0 ppm in both
spectra.
• Internal Standard: Because TMS is chemically inert, volatile, and has a simple spectrum (a
single sharp peak at 0 ppm), it is an ideal internal standard for calibrating the chemical shifts
of other compounds.
Properties:
• Inert and Non-Polar: TMS is chemically inert, meaning it does not react with most
compounds, and its non-polar nature ensures that it does not interfere with the sample
being analyzed.
• Single Peak: In 1H NMR, TMS shows a single peak because all the hydrogen atoms are in
equivalent environments, making it easy to distinguish from other signals.
Dr. R. C. Kulkarni | KLS GIT, BGM
Module III
Dr. R. C. Kulkarni | KLS GIT, BGM
Module III
13C chemical shifts
Since the organic chemist is interested primarily in the molecular carbon skeleton, a 13C NMR
spectrum yields structural information much more directly than a proton spectrum: quaternary
carbons, as those of many functional groups (CN, C=O, C=NR), are detectable. In some cases, 13C NMR
easily distinguishes between groups that might otherwise be confused, e.g. cyanide (110-120 ppm)
and isocyanide (135-150 ppm).
Dr. R. C. Kulkarni | KLS GIT, BGM
Module III
Dr. R. C. Kulkarni | KLS GIT, BGM
Module III
Dr. R. C. Kulkarni | KLS GIT, BGM
Module III
Dr. R. C. Kulkarni | KLS GIT, BGM
Module III
In general hydrogen bonded protons show higher value of chemical shift than the non-hydrogen
bonded protons. Due to the high electro-negativity of the atom to which proton is hydrogen bonded,
the electron cloud around the proton is decreased and thus causes deshielding of the proton. The
downfield shift of the absorption depends on the strength of the hydrogen bonding.
Greater is the degree of hydrogen bonding of proton, greater the downfield shift of a proton. This is
due to hydrogen bonding that the chemical shifts of OH, NH and SH protons vary over a wide range.
It is unpredictable as the extent of hydrogen bonding depends on concentration, temperature, solvent
and compound structure. The OH signals tend to move downfield at higher
compound concentration because of increased hydrogen bonding. Also on increasing temperature,
hydrogen bonding decreases and hence OH signals moves slightly upfield. Both OH and NH signals
move downfield in H-bonding solvents like DMSO or acetone.
3. Solvent Effects
Chloroform (CDCl3) is the most common solvent for NMR measurements. Other deuterium labelled
compounds, such as deuterium oxide (D2O), benzene-d6 (C6D6), acetone-d6 (CD3COCD3) and DMSO-
d6 (CD3SOCD3) are also used as NMR solvents. All these solvents vary considerably in their polarity
and magnetic susceptibility. Hence the NMR spectrum recorded in one solvent may be slightly
different from that recorded in other solvent. Hence it is important to mention the solvent used. The
NMR signals for protons attached to carbon are generally shifted slightly by changing solvent except
where significant bonding or dipole-dipole interaction might arise.
The changes in chemical shifts due to solvents changes are minor being on the order of ±0.1 ppm.
However, in some cases the changes are more. The aromatic solvents benzene and pyridine cause
changes in chemical shifts as large as 0.5 to 0.8 ppm compared to less magnetically active solvents like
Dr. R. C. Kulkarni | KLS GIT, BGM
Module III
chloroform or acetone. It is observed that the carbonyl groups form weak π–π collision complexes
with benzene rings that persist long enough to exert a significant shielding influence on nearby groups.
The second noteworthy change can be observed in the spectrum of tert-butanol in DMSO, where the
hydroxyl proton is shifted 2.5 ppm downfield from where it is found in dilute chloroform solution. This
is due to strong hydrogen bonding of the alcohol O–H to the sulfoxide oxygen, which not only deshields
the hydroxyl proton, but secures it from very rapid exchange reactions that prevent the display of
spin-spin splitting. Similar but weaker hydrogen bonds are formed to the carbonyl oxygen of acetone
and the nitrogen of acetonitrile.
Dr. R. C. Kulkarni | KLS GIT, BGM
Module III
Dr. R. C. Kulkarni | KLS GIT, BGM
Module III
Dr. R. C. Kulkarni | KLS GIT, BGM
Module III
Dr. R. C. Kulkarni | KLS GIT, BGM
Module III
Dr. R. C. Kulkarni | KLS GIT, BGM
Module III
Dr. R. C. Kulkarni | KLS GIT, BGM
Module III
Dr. R. C. Kulkarni | KLS GIT, BGM
Module III
Dr. R. C. Kulkarni | KLS GIT, BGM
Module III
Dr. R. C. Kulkarni | KLS GIT, BGM
Module III
Dr. R. C. Kulkarni | KLS GIT, BGM
Module III
Dr. R. C. Kulkarni | KLS GIT, BGM