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Sub topics of NMR spectroscopy
By
Hajira Mahmood
Ph.D. Scholar
Volume andConcentration Consideration
in NMR Spectroscopy
• Biomolecule samples.
• Prepare at least 0.05mm. higher than 1 mm , preferred as long as solution viscosity
does not increase significantly.
• Small molecule samples.
• About 3mm is sufficient for 13C direct NMR measurements, and10mM for 13C
measurements. If MW is 200, 10mg/500uL gives 100mM concentration.
• Note that for quantification, avoid preparing too high concentration which may
cause baseline and line shape issues.
Conti………
Sample volume should be minimum of 250 µL for a Shigemi tube
 160µL for a 3mm/5mm tube
 500 µL for a regular NMR tube.
Insufficient volume may cause bad shimming result, consequently spectra with
poor resolution & line shape.
In many cases the sample volume should weight more than the concentration
for a good NMR measurement.
The NMR sample should be homogeneous, free of air bubbles and insoluble
substances. High salt concentration and paramagnetic ions should be
avoided
Spectrometer setup
A typical NMR spectrometer setup consists of a superconducting
magnet, a probe, a console, and a computer.
The superconducting magnet is often a solenoid made from niobium
titanium alloy.
A large current flows around the loop, creating the strong continuous
magnetic field required to align the nuclei
The sample is placed in an NMR tube and dropped into the magnet on a
cushion of compressed air.
The strong magnetic field causes the nuclei to become aligned or
opposed to it.
Conti….
Deuterium Lock
The lock system keeps the spectrometer operating at a constant net
magnetic field.
Usually the liquid NMR uses deuterium solvent for stabilizing magnet
field (lock) & shimming (gradient shimming).
If you have to use samples without deuterium solvent, you are not able
to lock the field.
The magnet field may drift during the accumulation, resulted in broad
lines.
Probe Tunning for optimizing NMR signal
The probe is at the heart of an NMR spectrometer, in the centre of the
applied magnetic field. The probe holds the:
Sample and sample spinning apparatus
Radio frequency (RF) coils
Temperature controller
Detector coils
Conti…
The term tuning involves two connected activities:
Tuning the coil to the required frequency
Matching the coil to the correct impedance ensuring its efficient
operation
The circuit requiring tuning is composed of inductive (L) and
capacitive (C) elements and the L and C values determine the
frequency of the RF circuit.
A circuit is tuned by adjusting the capacitor values until the required
frequency is reached.
Sample Tune Placement
An NMR tube is a thin glass walled tube used to contain samples in nuclear magnetic
resonance spectroscopy.
Typically NMR tubes come in 5 mm diameters but can also range from 10 mm to
3 mm samples.
 It is important that the tubes are uniformly thick & well-balanced to ensure that NMR
tube spins at a regular rate (i.e. that they do not wobble usually about 20 Hz in an NMR
spectrometer.
For 1H, 0.5 - 5.0 mg of the sample is sufficient.
For high molecular weight samples, more concentrated solutions are recommended.
However, too concentrated solution leads to lower resolution due to saturation and/or
increased viscosity.
For 13C and other nuclei five times the concentration of 1H is recommended (20-100
mg.
sub topics of NMR 2.pptx

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sub topics of NMR 2.pptx

  • 1.
  • 2. Sub topics of NMR spectroscopy By Hajira Mahmood Ph.D. Scholar
  • 3. Volume andConcentration Consideration in NMR Spectroscopy • Biomolecule samples. • Prepare at least 0.05mm. higher than 1 mm , preferred as long as solution viscosity does not increase significantly. • Small molecule samples. • About 3mm is sufficient for 13C direct NMR measurements, and10mM for 13C measurements. If MW is 200, 10mg/500uL gives 100mM concentration. • Note that for quantification, avoid preparing too high concentration which may cause baseline and line shape issues.
  • 4. Conti……… Sample volume should be minimum of 250 µL for a Shigemi tube  160µL for a 3mm/5mm tube  500 µL for a regular NMR tube. Insufficient volume may cause bad shimming result, consequently spectra with poor resolution & line shape. In many cases the sample volume should weight more than the concentration for a good NMR measurement. The NMR sample should be homogeneous, free of air bubbles and insoluble substances. High salt concentration and paramagnetic ions should be avoided
  • 5. Spectrometer setup A typical NMR spectrometer setup consists of a superconducting magnet, a probe, a console, and a computer. The superconducting magnet is often a solenoid made from niobium titanium alloy. A large current flows around the loop, creating the strong continuous magnetic field required to align the nuclei The sample is placed in an NMR tube and dropped into the magnet on a cushion of compressed air. The strong magnetic field causes the nuclei to become aligned or opposed to it.
  • 7. Deuterium Lock The lock system keeps the spectrometer operating at a constant net magnetic field. Usually the liquid NMR uses deuterium solvent for stabilizing magnet field (lock) & shimming (gradient shimming). If you have to use samples without deuterium solvent, you are not able to lock the field. The magnet field may drift during the accumulation, resulted in broad lines.
  • 8. Probe Tunning for optimizing NMR signal The probe is at the heart of an NMR spectrometer, in the centre of the applied magnetic field. The probe holds the: Sample and sample spinning apparatus Radio frequency (RF) coils Temperature controller Detector coils
  • 9. Conti… The term tuning involves two connected activities: Tuning the coil to the required frequency Matching the coil to the correct impedance ensuring its efficient operation The circuit requiring tuning is composed of inductive (L) and capacitive (C) elements and the L and C values determine the frequency of the RF circuit. A circuit is tuned by adjusting the capacitor values until the required frequency is reached.
  • 10. Sample Tune Placement An NMR tube is a thin glass walled tube used to contain samples in nuclear magnetic resonance spectroscopy. Typically NMR tubes come in 5 mm diameters but can also range from 10 mm to 3 mm samples.  It is important that the tubes are uniformly thick & well-balanced to ensure that NMR tube spins at a regular rate (i.e. that they do not wobble usually about 20 Hz in an NMR spectrometer. For 1H, 0.5 - 5.0 mg of the sample is sufficient. For high molecular weight samples, more concentrated solutions are recommended. However, too concentrated solution leads to lower resolution due to saturation and/or increased viscosity. For 13C and other nuclei five times the concentration of 1H is recommended (20-100 mg.