Variable Temperature to Improve NMR Resolution

Many millions of dollars have been spent on high field NMR magnets to improve both sensitivity and chemical shift dispersion.  Many younger NMR users have had the good fortune to use only high field spectrometers where chemical shift resolution is often not an issue.  These users are not familiar with some of the "tricks" used to improve resolution which were needed on lower field instruments where chemical shift resolution was frequently a problem.  With the current helium shortage and the increasing popularity of low field permanent magnet spectrometers, these "tricks" will again become more and more common.  Among them are; the use of paramagnetic chemical shift reagents, the use of aromatic solvents or solvent mixtures and the use of variable temperature.  In this post, I would like to demonstrate the incredible power of simply changing the temperature at which the NMR data are collected.

The ¹H chemical shift is a sensitive parameter related to the conformation of a molecule.  In solution, small molecules may adopt a number of conformations whose populations depend on the potential energy profile.  Furthermore, the molecules are often in fast exchange between the available conformations and the observed chemical shift is the weighted average chemical shift of all of the conformations present.  As the temperature is changed, the populations of conformations are altered and the observed average chemical shift value may change.  The changes in chemical shifts at different temperatures are often enough to resolve resonance which may have overlapped with one another at room temperature.

The chemical shift of exchangeable protons ( -OH, -NH or NH₂) depends dramatically on the degree of both inter-molecular and intra-molecular hydrogen bonding.  When molecules with exchangeable protons are dissolved in aprotic solvents, one is often able to observe the exchangeable protons as well as their associated J couplings.  Such is the case with sucrose dissolved in DMSO-d₆ where all of the -OH protons can easily be observed.  When the temperature is changed, the populations of available conformations change and the degree of intra-molecular hydrogen bonding is affected with dramatic changes in the chemical shifts of the -OH resonances.  The figure below shows the anomeric and -OH region of the 500 MHz ¹H NMR spectrum of sucrose in DMSO-d₆ collected as a function of temperature.

All of the protons can be assigned with standard 2D NMR methods.  As the temperature is increased, the anomeric proton (1) moves to higher frequencies while the -OH protons (2-9) all move to lower frequencies to different extents.  Note that the resonances in the highlighted region of the spectrum at 21°C are overlapped with one another but at higher temperatures are fully resolved.  The resolution has increased by simply increasing the temperature.

Resolution During a Liquid Nitrogen Refill

Just as it is not advisable to collect high resolution NMR data during (and shortly after) a liquid helium refill, it is similarly not advisable to collect data during a liquid nitrogen fill. The figure below illustrates this point. A 300 MHz NMR spectrometer was set up to collect an NMR spectrum every minute before, during and after a liquid nitrogen refill. The resolution deteriorates as soon as the one-way valves are removed from the nitrogen cryostat. One can see that the resolution during the fill is much worse than before the fill. Even 60 minutes after the fill, the magnet has not fully recovered the resolution attained before the fill. The last spectrum of the series was collected after re-shimming the magnet and is comparable to the spectrum collected before the fill. The resolution profile during a fill may be different for every magnet. For this particular magnet, it is advisable not to collect high resolution data for at least 90 minutes after a nitrogen refill.

Resolution During a Liquid Helium Refill

Students sometimes ask whether or not they can collect NMR data during a liquid helium refill. The answer depends on the type of NMR experiment. For most solids experiments, where high resolution ( <~ 100 Hz) is not required, one can often collect data without a problem. In experiments for liquids, where high resolution is almost always required, the quality of data are affected drastically by the disturbance from the liquid helium refill and it is not advisable to collect data during this time. Furthermore, after a liquid helium refill, the magnet must recover in order to obtain the same resolution prior to the fill. The figure below illustrates this point. A 300 MHz NMR spectrometer was set up to collect an NMR spectrum every minute before, during and after a liquid helium refill. One can see that the resolution during the fill is much worse than before the fill. Even 15 minutes after the fill, the magnet has not fully recovered the resolution attained before the fill. The last spectrum of the series was collected after re-shimming the magnet and is comparable to the spectrum collected before the fill.
The resolution profile during a fill will probably be slightly different for every magnet. For this particular magnet, it is advisable not to collect high resolution data for at least 30 minutes after a helium fill.

Nitrogen Pressure and Spectral Resolution

Super-conducting NMR magnets are cooled with liquid helium. The boil off rate of the liquid helium is minimized by surrounding it with a high vacuum and a liquid nitrogen cryostat. The liquid nitrogen cryostat may either be vented directly to the atmosphere or it may be maintained at a pressure slightly above atmospheric pressure with the use of pressure relief valves. NMR users should be aware that the homogeneity of an NMR magnet is affected drastically by the pressure in the liquid nitrogen cryostat. The left-hand panel of the figure below shows a well resolved 400 MHz ¹H NMR spectrum of the methyl triplet of ethylbenzene in a well shimmed magnet where the nitrogen cryostat was maintained slightly above atmospheric pressure with pressure relief valves for several days. The spectrum in the right-hand panel was run under identical conditions after the cap to the nitrogen fill port was removed, relieving the pressure in the nitrogen cryostat.