Nuclear Magnetic Resonance (NMR) Spectroscopy: Insights into Molecules & Diagnostics
Group 7, Lais P.C Alexandra P
Introduction
Basic principles of the general technique (chromatographic
spectroscopic)
What is spectroscopic: spectroscopy, is the study of the absorption and emission of light and other
radiation by matter, as related to the dependence of these processes on the wavelength of the
radiation. More recently, The definition has been expanded to include the study of the interactions
between particles such as electrons, protons, and ions, as well as their interaction with other
particles as a function of their collision energy.
Basic principles of the selected method (GC, HPLC, SEC, IEC, TLC, UV-VIS, IR,
AA, MS)
Nuclear Magnetic Resonance (NMR) spectroscopy is a powerful analytical technique used to study
the molecular structure and dynamics of organic and inorganic compounds. It provides information
about the arrangement of atoms in a molecule, their connectivity, and the environment they are in.
NMR is based on the interaction between the nuclei of certain atoms and an external magnetic field.
Nuclei with an odd number of protons or neutrons (i.e., having a nuclear spin) possess a magnetic
moment and can absorb and emit radiofrequency (RF) energy when placed in a strong magnetic
field and subjected to RF radiation. Here are the basic principles of NMR spectroscopy:
NMR Spectrometer:
An NMR spectrometer is the instrument used to measure the absorption and emission of
electromagnetic radiation by the nuclei. It consists of a strong magnet to create a uniform magnetic
field, A radiofrequency (RF) source to generate the excitation pulses, and a detector to measure the
emitted signals.
Instrumentation:
NMR spectrometers consist of a strong magnet, a sample holder, and RF equipment. The most
common nuclei used in NMR are hydrogen (^1H) and carbon-13 (^13C), due to their high natural
abundance and sensitivity.
Nuclear Spin:
Atomic nuclei with an odd number of protons or neutrons possess an intrinsic property called spin.
This spin creates a magnetic moment that interacts with an external magnetic field.
Chemical Shift:
The chemical shift is a crucial parameter in NMR spectroscopy. It is the difference in resonance frequency of a nucleus in a
compound compared to a reference compound (often tetramethylsilane, TMS). Chemical shifts are measured in parts per million
(ppm) and provide information about the local electronic environment of the nuclei.
Spin-Spin Coupling (J-Coupling):
Adjacent nuclei with different spins can influence each other's resonance frequencies through a
process called spin-spin coupling. This interaction results in the splitting of NMR signals into
multiplets, providing insights into the connectivity of neighboring atoms.
Nuclear Magnetic Resonance:
When a sample containing nuclei with spin is placed in a strong magnetic field, the nuclear
magnetic moments align either with or against the external field. This alignment creates a slight
excess of nuclear spins in the lower-energy state
Pourmodheji, H., Ghafar-Zadeh, E., & Magierowski, S. (2016). A Multidisciplinary Approach to High Throughput Nuclear Magnetic Resonance
Spectroscopy. Sensors, 16(6), 850. MDPI AG. Retrieved from http://dx.doi.org/10.3390/s16060850rrafo
Description of the application of interest
Absorption and Emission of Electromagnetic Radiation:
Study Objectives:
or
The objective of this study was to use nuclear magnetic resonance (NMR) spectroscopy to
By applying a specific frequency of electromagnetic radiation (radio waves) to the sample, the nuclei can absorb
identify and analyze metabolic alterations associated with chronic kidney disease (CKD)
energy and transition to a higher-energy state. This absorption occurs at a frequency that matches the energy
progression. The study aimed to provide a better understanding of the metabolic changes
difference between the two spin states, determined by the external magnetic field strength.
occurring in patients with CKD and potentially identify target pathways that could be modified
to slow down the disease's progression.
Resonance Condition:
The frequency at which the nuclei absorb electromagnetic radiation is known as the resonance frequency. It depends
Application of the Method:
on the strength of the magnetic field and the type of nucleus being observed. The resonance frequency can be
The researchers employed 1H NMR spectroscopy combined with multivariate analysis to
calculated using the Larmor equation: ω = γB₀, where ω is the resonance frequency, γ is the gyromagnetic ratio of the
analyze serum metabolites in patients with various stages of CKD. They compared the
nucleus, and B₀ is the magnetic field strength.
metabolite profiles of CKD patients with those of healthy controls using NMR spectroscopy and
explored the differences in metabolite concentrations and pathways.
Integration:
The area under an NMR signal is proportional to the number of equivalent nuclei giving rise to that signal. This
Significant Findings/Results:
integration can provide information about the relative number of different types of nuclei in a molecule.
The 1H NMR spectroscopy analysis combined with multivariate analysis revealed significant
differences in the metabolite profiles between CKD patients and healthy controls. The study
2D NMR Techniques: identified 15 endogenous metabolites that differed between the two groups. Metabolic
Two-dimensional NMR techniques, such as COSY (correlation spectroscopy), NOESY (nuclear Overhauser effect
Pathway Analysis (MetPA) further demonstrated significant intergroup differences in five
spectroscopy), and HSQC (heteronuclear single quantum coherence), allow for the identification of connectivities
potential target pathways and 14 metabolites. The discriminant models based on these
between different nuclei in a molecule, enabling the determination of complex molecular structures.
findings indicated that CKD patients and healthy controls could be differentiated based on
their metabolic fingerprints. The results suggested that CKD and its progression have an
Nuclear Spin Relaxation: impact on selected metabolic pathways.
After absorbing energy, the nuclei return to their lower-energy state, releasing the absorbed energy as electromagnetic
radiation. This relaxation process occurs through two mechanisms: spin-lattice relaxation (T₁) and spin-spin relaxation
Conclusion:
(T₂). T₁ represents the time taken for the nuclear spin to recover parallel to the magnetic field, while T₂ represents the The study demonstrated the application of 1H NMR spectroscopy in identifying metabolic
time for the loss of phase coherence between spins. disorders associated with CKD. The findings provided valuable insights into the metabolic
alterations occurring in CKD patients and highlighted specific target pathways that may be
Applications: modified to slow down the disease's progression. The use of NMR spectroscopy for rapid
NMR spectroscopy is widely used in organic chemistry, biochemistry, drug discovery, and material science to determine detection of endogenous metabolites offers the potential for monitoring and understanding
molecular structures, study reaction mechanisms, analyze molecular dynamics, and investigate intermolecular CKD-related metabolic changes, potentially leading to improved prevention and
interactions. management strategies.
Overall, NMR spectroscopy provides detailed information about the structure, dynamics, and interactions of molecules, Summary of method and
making it an indispensable tool in various scientific disciplines.
application of interest
The study utilized 1H NMR spectroscopy to
analyze serum samples from CKD patients and
healthy controls. The technique allowed for the
detection and quantification of endogenous
metabolites in the samples. The obtained NMR
spectra were subjected to multivariate analysis
to identify significant differences in the
metabolite profiles between the two groups. This
approach provided insights into the metabolic
alterations associated with CKD and identified
target pathways that could be potentially
modified to slow down the progression of the
disease.
FollowShiqi WangMSCA Early Stage Researcher at Imperial College London. (2014, October 28). Nuclear magnetic
resonance spectroscopy (NMR spectroscopy) - ITN snal - marie curie initial trainin
g network. ITN SNAL - Marie Curie Initial Training Network - Smart Nano-objects for Alteration of Lipid bilayers
. https://itn-snal.net/2014/10/nuclear-magnetic-resonance-spectroscopy/
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Mika, A., Wojtowicz, W., Ząbek, A., Młynarz, P., Chmielewski, M., Sledzinski, T., & Stepnowski, P.
(2018). Application of nuclear magnetic resonance spectroscopy for the detection of metabolic
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Mika, A., Wojtowicz, W., Ząbek, A., Młynarz, P., Chmielewski, M., Sledzinski, T., & Stepnowski, P. (2018). Application of nuclear magnetic resonance spectroscopy for the detection of metabolic Biomedical Analysis, 149, 1–8. https://doi.org/10.1016/j.jpba.2017.10.037
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