Fourier Transform NMR (FT-NMR)

Rutuja Dattatray Chougale,

Research Scholar,
M. Pharm Semester I,
Tatyasaheb Kore College of Pharmacy, Warananagar.

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 Introduction
Nuclear magnetic resonance (NMR) spectroscopy is a very important spectroscopic method
that provides crucial information relating to chemical structure of molecule.
NMR is a technique which gives information about the type and number of magnetically
distinct protons in a molecule.
Nuclear magnetic resonance spectroscopy is a spectroscopic method in which, under
appropriate conditions, a sample placed in a magnetic field, absorbs electromagnetic
radiation in the radiofrequency (rf) region.
The frequency of absorption is governed by the characteristics of the sample and it is a
function of certain nuclei constituting the sample.
Example : Proton NMR (1H-NMR) spectroscopy gives information about hydrogen nuclei,
and one can determine number of distinct types of hydrogen nuclei as well as information
regarding the nature of immediate chemical environment for each type of 1H nucleus.
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Why Fourier Transform NMR?
In a CW-NMR instrument operating on field sweep or frequency sweep modes, the
nuclei undergo excitation one at a time.
In the case of 1H-NMR, chemically distinct type of protons (phenyl, vinyl, methyl
and so on) are excited individually and come to resonance in succession
independently of other nuclei. The scanning continues till all varying types of
protons have come to resonance one by one.
The NMR spectrum obtained therefore, consists of a series of peaks plotted on the
abscissa (x-axis) corresponding to variation in the field or frequency.
As we look at first one type of hydrogen and then another scanning until all of the
types have come into resonance.
A disadvantage with this mode of operation is that the spectrum takes several
minutes to be recorded.
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Pulsed Fourier Transform NMR Spectrophotometer
An alternative approach followed very commonly in modern sophisticated
NMR instrument is the pulsed Fourier Transform NMR.
In this method, all transitions are stimulated simultaneously within a
fraction of a second by irradiating the sample (at a fixed B0) with a
powerful but short burst of rf energy (called a pulse) (approximately 10
s) that contains the complete range of frequencies required to cover the
varying types of protons (e.g., spread around 90 MHz in an instrument
with a 2.1 T magnet or around 100MHz for a 2.3 T instrument).

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 Fourier Transform NMR Spectroscopy
The Fourier Transformation is the basic mathematical calculation
necessary to convert the data in time domain (interferogram) to frequency
domain (NMR Spectrum).
i.e., time domain  Intensity v/s Time.
Frequency domain  Intensity v/s Frequency.
It was developed by JEAN BAPTISE JOSEPH FOURIER.

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Pulsed Fourier Transform NMR Spectrophotometer

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 Theory of FT-NMR
When magnetic nuclei are placed in a magnetic field and irradiated with a
pulse of radio frequency close to their resonant frequency, the nuclei
absorb some of the energy and precess like little tops at their resonant
frequencies.

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 Theory of FT-NMR
This precession of many nuclei at slightly different frequencies produces a
complex signal that decays as the nuclei loses the energy they had gained
from the pulse. This signal is called as free induction decay (FID) or
transient ,it contains all the information needed to calculate a spectrum.
The free induction decay can be recorded by a radio receiver and a
computer in 1-2 seconds and many FIDs can be averaged in few minutes.
A computer converts the average transients into a spectrum.

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 Pulsed Fourier Transform NMR Spectrophotometer
The powerful pulse excites all the magnetic nuclei in the molecule simultaneously
as they absorb their own respective frequencies out of the pulse and all the signals
are collected at the same time with a computer where these frequencies couple to
give beats.
On discontinuation of the pulse, the excited nuclei begin to relax to their original
spin states by losing their excitation energy. In this process, the nuclei re-emit the
absorption energies and coupling energies a electromagnetic radiation.
As one molecule contains several different types of 1H nuclei (protons), many
different frequencies of electromagnetic radiation are simultaneously emitted. This
emission is called a free-induction decay (FID) signal.
The intensity of FID rapidly decays with the time as all the absorbed frequencies
are eventually emitted by the excited nuclei.

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 Pulsed Fourier Transform NMR Spectrophotometer
The observed FID is an interference signal between the radiofrequency
source and the frequency emitted by the excited protons.
This output is digitized in a computer and individual frequencies due to
different nuclei are extracted out from the interference pattern
(interferogram) by on a computer by a mathematical method called a
Fourier-transform analysis.
The Fourier transform separates the FID into its sine or cosine wave
components.
The wavelength is given as:
 = 1/nucleus - pulse

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 Schematic diagram of FT-NMR spectrometer with a
super-conducting magnet

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 Instrumentation

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 Components of FT-NMR
A radio transmitter coil that produces a short powerful pulse of radio waves.
A powerful magnet that produces strong magnetic fields.
The sample is placed in a glass tube that spins so the test material is subject to
uniform magnetic field.
A radio receiver coil that detects radio frequencies emitted as nuclei relax to a
lower energy level.
A computer that analyses and record the data.
In Fourier transform NMR instruments, the tube fits within the bore of the
superconducting magnet or a solenoid and spins about the z-axis which is vertical.
In other words, the probe is parallel to the z-axis of the magnet which is cooled
with liquid helium surrounded by liquid nitrogen in a large Dewar flask.

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 Sensitivity: Signal to noise ratio
Sensitivity of NMR technique is assessed in terms of the signal to noise ratio.
Noise refers to the randomelectronic signals usually visible as baseline
fluctuations.

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 Sensitivity: Signal to noise ratio
The signal to noise ratio (S/N) indicates the sensitivity of an FT-NMR
experiment
S/N = NsT2exc (Bdet)1/3(ns)1/2/T
Here, Ns = Number of spins in the system or sample amount
T2 = Transverse relaxation time(determine line width)
exc and det = Magnetogyric ratio of excited and detected nuclei
ns = Number of scans
B = Strength of external magnetic field
T = Temperature of the sample

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 Modifications in FT-NMR
In most labs, the cheapest and easiest route is to increase the number of scans n s,
but this increases the experiment time proportionately (Commonly used method in
13C NMR).

Routinely, sample tubes with 5mm (outer diameter) are available which use about
10 mg of sample dissolved in about 0.5 mL solvent. For greater sensitivity,
microprobes are available with outer diameters 1.0mm, 2.5mm or 3.0mm. As low
as 100 ng of an average molecular weight compound may be analyzed in a 1.0 mm
tube (volume 5 L) in a 600 MHz instrument.
The development of cryogenically cooled probes (cryo-probes) has significantly
decreased sample amount requirements. These have built in first-stage receivers
and rf coils which are cryogenically cooled (approx. 20K), and result in almost 4
times improvements in S/N values.

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 Modifications in FT-NMR
Higher field instruments (higher B) conceptually provide higher
sensitivity. For a fixed concentration (Ns), sample requirement becomes
almost 2.8 times lesser on a 600 MHz instrument compared to 300 MHz
instrument in order to obtain spectra with identical S/N :
S/N = Ns 600/Ns 300 = (600/300)1/3 2.8
Thus, if smaller sample amount is available, one should go in for highest
field instruments with the smallest possible diameter cryo-probe. A cryo-
capillary flow microprobe can be used to dissolve few nanograms of the
sample in approximately 1L of the solvent for highest sensitivity.

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 Advantages of FT-NMR
FT-NMR is much faster than CW-NMR method. An entire spectrum is
recorded, digitized and transformed within a few seconds. In comparison,
a CW spectrum takes 5 to 10 minutes.
The signal stand out clearly with low background noise.
With a computer and fast measurement, FID signal can be repeated and
averaged with a repetition every 2 seconds. As noise is random, its
intensity does not increase when repeated measurements of the spectrum
are added together. Hence, S/N ratio improves as number of scans ‘n’ is
increased.
S/N = fn

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 Advantages of FT-NMR
Pulsed NMR is more sensitive and weaker signals can be measured by this method.
The possibility of repeated FID measurements is an enormous advantage in cases
where signals are weak in intensity and which have a great amount of noise
associated with them.
Due to greater sensitivity, much lower concentrations of samples can be analyzed
compared to CW-NMR instruments. This is particularly useful for biological
samples where only microgram quantities may be available for analysis.
Sparingly soluble compounds can also be analyzed due to high sensitivity of the
techniques as even small number of nuclei in solution can give reasonably sharp
peaks.
This is useful for NMR studies on nuclei with low abundance and small magnetic
moments (e.g., 13C, 15N, 17O).

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