Nuclear Magnetic Resonance (NMR) Analysis

Nuclear Magnetic Resonance (NMR) Analysis 2019-02-15T18:55:28+00:00

NMR spectroscopy is a powerful analytical technique that can be used for a wide variety of solid and liquid materials.  Specific NMR active nuclei within a sample can be observed, which confers a unique level of selectivity for NMR spectroscopy.  AMRI routinely uses both solid-state and liquid-state cGMP NMR spectroscopy to characterize a wide variety of materials from small inorganic compounds to large molecules.  Specific NMR capabilities to analyze nuclei like 19F in the both solids and liquids are beneficial because fluorine is present in about 20% of pharmaceutical drugs.  Advanced NMR capabilities for analyzing large molecules are important because biologics are the fastest growing area of pharmaceutical drug development.

NMR Applications:

AMRI provides an in-depth and broad experience with NMR spectroscopy of small molecules, synthetic polymers, and biologics.  Some of the most common liquid and solid state NMR applications performed for our customers are summarized below.

  • 1D ¹H and ¹³C NMR spectroscopy with 19F, 31P, 29Si available as routine analyses.
  • 2D NMR analyses such as DQF-COSY (Figure 1), NOESY, TOCSY, HMQC, HSQC, and HMBC with resonance assignments to provide detailed information on chemical structure and conformation in solution.

  • USP/NF and EP compendial NMR methods
  • Qualitative and quantitative NMR (qNMR) method development and validation for all available liquids and solids NMR techniques. Most of our validated methods are used for clinical or commercial lot release or stability studies.
  • Comparability studies of biologics based on the sensitive and accurate fingerprint of the molecular conformation extracted from the NMR data
  • Identification and quantification of known and unknown impurities (solids and liquids)
  • Characterization of polymorphs, solvates, salts, co-crystals, and amorphous solids
  • Chemical structure identification (solids and liquids)
  • Analysis of formulations (solids and liquids)
  • Analysis of stereoisomers (solids and liquids)
  • Confirm the number of molecules in the asymmetric unit (solids)
  • Chemical exchange analysis (solids and liquids)
  • Molecular motion analysis (solids and liquids)
  • Conformational/structural analysis (solids and liquids).
  • HR-MAS NMR spectroscopy of semisolids and suspensions
  • 3D chemical structure determination of small molecules and biologics using NMR restrained molecular dynamics (MD) simulations

NMR Service Benefits at AMRI:

  • Extensive experience with NMR spectroscopy of small molecules and large molecules (polymers, unmodified and modified peptides, proteins, DNA, RNA, lipids)
  • Higher resolution and sensitivity with modern liquids and solids NMR spectrometers and probes
  • Customer-selected data interpretation levels (very basic to very detailed) for maximum value and flexibility
  • Higher quality 2D NMR spectra through advanced pulse sequences and data processing methods
  • cGMP compliance with FDA and EMA regulations to ensure the highest quality data, interpretation, and experimental consistency

Our equipment includes 300 MHz to 600 MHz NMR spectrometers some of which are capable of analyzing both solids and liquids.

Liquid-state 400 MHz NMR (Figure 2):

  • 5 mm double resonance pulsed field gradient (PFG) probe for high sensitivity on the X and Y channels (useful for typical small organic molecules and biologics).  Observe 1H (~400 MHz) or 19F (~376 MHz) on the high band channel and 15N (~40 MHz) to 31P (~162 MHz) on the low band channel.
  • 5 mm inverse detection PFG probe (useful for biologics and typical small organic molecule samples).  Observe 1H (~400 MHz) or 19F (~376 MHz) on the high band channel and 15N (~40 MHz) to 31P (~162 MHz) on the low band channel.
  • 5 mm dual broadband PFG probe (ideal for organic and inorganic materials to observe 13C, 15N, and other lower frequency nuclei or those nuclei with low natural abundance). Observe 1H (~400 MHz) or 19F (~376 MHz) on the high band channel and 15N (~40 MHz) to 31P (~162 MHz) on the low band channel.
  • Z-axis gradient capabilities for performing many of the newest pulsed field gradient experiments.
  • Wide range of 2D NMR spectroscopic techniques available for structural elucidation.
  • Variable temperature capabilities from −80 °C to 130 °C.
  • Sample quantities in ~0.5 mL solvent:
  • 1H or 19F – 0.1-5 mg, 31P – 10-20 mg, 13C – 20-50 mg

Liquid-state 600 MHz NMR:

  • 5 mm double resonance pulsed field gradient (PFG) cryoprobe for very high sensitivity on the X and Y channels for the most challenging chemical structure studies of minor impurities and large biological molecules.  Observe 1H (~600 MHz) or 19F (~564 MHz) on the high band channel and 15N (~60 MHz) to 31P (~243 MHz) on the low band channel.
  • 5 mm double resonance pulsed field gradient (PFG) standard probe for high sensitivity on the X and Y channels for the typical chemical structure determination of impurities and large biological molecules.  Observe 1H
  • (~600 MHz) or 19F (~564 MHz) on the high band channel and 109Ag (~28 MHz) to 31P (~243 MHz) on the low band channel.
  • Z-axis gradient capabilities for performing any of the newest pulsed field gradient experiments.
  • Wide range of 2D NMR spectroscopic techniques available for structural elucidation.
  • Variable temperature capabilities from −150 °C to 150 °C.
  • Very small sample quantities in ~0.5 mL solvent:
  • 1H or 19F – 0.01-5 mg, 31P – 1-10 mg, 13C – 1-20 mg

Solid-state 400 MHz NMR (Figure 3):

  • Agilent T3 narrow bore double and triple resonance 4 mm solid-state NMR probes.
  • Doty Scientific 4 mm HF probe specifically designed for simultaneous 1H decoupling while detecting 19F or the reverse.
  • Doty Scientific 4 mm HFX probe specifically designed for simultaneous 1H decoupling while detecting 19F or X nuclei or 19F decoupling while observing 1H or X.
  • Low frequency: Observe any NMR active nucleus with a resonance frequency between 15N (~40 MHz) and 31P (~162 MHz).
  • High Frequency: Observe or decouple 1H (~400 MHz) and 19F (~376 MHz) nuclei.
  • MAS and CP/MAS available up to 18 kHz spinning speed (with modern phase modulated decoupling and/or TOSS spinning sideband suppression) to offer the highest data quality.
  • Simultaneous 1H and 19F decoupling while observing on the X-channel, and 1H or 19F decoupling while observing 19F or 1H.
  • 2D correlation spectroscopy of common nuclei (1H, 13C, 15N, 19F, 29Si, 31P).
  • 2D MQMAS or STMAS of quadrupolar nuclei.
  • HR-MAS for semisolids and suspensions
  • Variable temperature capabilities from −75 °C to 100 °C
  • Sample quantities: 40-50 mg (optimal), 5-10 mg (possible)
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