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Chemical Shift: Internal References and Solvent Effects01:17

Chemical Shift: Internal References and Solvent Effects

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In an NMR sample, precise measurement of the absolute absorption frequencies of nuclei is difficult. A standard internal reference compound is added, and the frequency difference between the reference signal and sample signals is measured.
The internal reference compound generally used in NMR spectroscopy is tetramethylsilane (TMS). TMS is preferred because it is chemically inert, soluble in NMR solvents, and easily removable. Also, the highly shielded methyl protons in TMS yield an intense...
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Heating a crystalline solid increases the average energy of its atoms, molecules, or ions, and the solid gets hotter. At some point, the added energy becomes large enough to partially overcome the forces holding the molecules or ions of the solid in their fixed positions, and the solid begins the process of transitioning to the liquid state or melting. At this point, the temperature of the solid stops rising, despite the continual input of heat, and it remains constant until all of the solid is...
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Updated: Jan 18, 2026

Cryogenic Sample Loading into a Magic Angle Spinning Nuclear Magnetic Resonance Spectrometer that Preserves Cellular Viability
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Millisecond-time-scale controlled freeze-quench for solute-intermediate analysis by solid-state NMR.

Ieva Goldberga1, Trevys Chanal1, Tristan Georges1

  • 1Sorbonne Université, CNRS, Chimie de la Matière Condensée de Paris, LCMCP, F-75005 Paris, France. thierry.azais@sorbonne-universite.fr.

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Summary

This study introduces a novel cryo-fixation method for studying fast biomineralization reactions. The technique uses solid-state NMR to capture transient prenucleation species, enabling detailed analysis of crystallization intermediates.

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Area of Science:

  • Biomineralization
  • Materials Science
  • Analytical Chemistry

Background:

  • Biomineralization involves complex crystallization pathways with transient intermediates.
  • Characterizing these intermediates is challenging due to their rapid formation and dynamic nature.
  • Prenucleation species are particularly difficult to study because of their solubility, small size, and instability.

Purpose of the Study:

  • To develop an innovative method for trapping and characterizing reactive intermediates in fast crystallization processes.
  • To enable time-resolved analysis of non-classical nucleation and growth pathways.
  • To apply the method to study early-stage calcium phosphate crystallization.

Main Methods:

  • Developed a cryo-fixation technique by spraying solutions into liquid isopentane at -145 °C.
  • Adapted sample preparation for low-temperature solid-state Nuclear Magnetic Resonance (NMR) spectroscopy.
  • Utilized a stopped-flow device for millisecond-timescale control of reaction aging.

Main Results:

  • Successfully vitrified phosphate solutions and analyzed them using low-temperature 31P solid-state NMR.
  • Distinguished different phosphate species based on pH-dependent chemical shift anisotropy patterns.
  • Observed early stages of amorphous calcium phosphate nucleation within 20 milliseconds of reaction time.

Conclusions:

  • The cryo-fixation and low-temperature solid-state NMR method allows for the study of transient species in fast, out-of-equilibrium aqueous reactions.
  • This technique preserves the native environment of intermediates, providing insights into biomineralization processes.
  • The methodology is applicable to a wide range of aqueous reactions beyond biomineralization.