The MSU NMR facility offers a full range of NMR related services applicable to single sample or high-throughput demands. Services include structure determination, diffusion coefficient measurements, homogeneity/mixture analysis, ligand binding/screening, kinetics and reaction monitoring, dynamics analysis, quantitative analysis, purity/impurity analysis, quality control, unknown/natural product isolation and analysis, HPLC purification, metabolomics analysis (serum, plasma, urine, tissue, cells, and media), education (lab sections and tours) and training (operation and data analysis). The NMR facility is managed full time by Dr. Brian Tripet who has >12 years NMR experience and provides expertise and  help to users with NMR experiment selection, NMR experiment set-up, sample analysis, data analysis and any problems encountered during use, data transfer and data management.

 

A detailed description of present facility services are listed below.

Small Molecule Metabolomics Determination

  • 1D NMR experiments: 1H-NOSEYPR1D and 1H-ZGESGP.
  • 2D NMR experiments: 1H-1H TOCSY.

Small Molecule Structure Determination

  • 1D NMR experiments: 1H, 13C, 19F, 31P, 2H, DEPT(45,90,135,q), APT, selNOE, selCOSY, selTOCSY, T1, T2, H/D-exchange, other X nuclei.
  • 2D NMR experiments: COSY, TOCSY, HMBC, HETCOR/COLOC, HMQC, HSQC, NOESY, ROESY, ADEQUATE/INADEAQUATE.
  • 3D NMR experiments: HMQC-COSY, HSQC-TOCSY, HMBC1H, 13C, 31P-correlations.

Small Molecule Related Experiments

  • Diffusion NMR (DOSY)-diffusion coefficient measurements, homogeneity/mixture analysis.
  • Saturation-Transfer-Difference (STD) NMR –a simple fast method for ligand screening and characterization of small molecule-protein binding.

Biomolecular/Protein Structure Determination and Analysis

  • Full capabilities for acquiring all necessary 1D, 2D, 3D and nD NMR experiments required for the three-dimensional solution structure determination of large macromolecules such as proteins (NMR experiments include HN-HSQC, HNCA, HNCO, CBCA(CO)NH, NH(CA)NNH, and others).
  • Dynamics, T1, T2, NOE relaxation studies – for analysis of dynamic changes associated with mutations in protein sequences, ligand binding and macromolecular motions.
  • Ligand Binding studies – STD (Saturation-Transfer-Difference) and HSQC titration analysis for elucidating interacting ligands or binding site.

Quantitative Analysis

  • Quantitative analysis of known and unknowns compounds (e.g. juice sugars/carbohydrates, saturation level of lipids and fats in edible oils).
  • Internal and pre-made capillary inserts for standardization and normalization.

LC-SPE-NMR-MS

(a hyphenated LC - NMR spectroscopy system with enhanced sample recovery combined with real-time MS analysis and high-through-put capabilities).

  • Analysis of known and unknowns following reversed-phased or HILIC chromatography.
  • Analytes can be trapped as fraction or on solid-phase-support (SPE) cartridges for drying, elution with deuterated NMR solvents and NMR analysis.

HPLC Purification

  • Agilent 1100 HPLC and columns (reversed phased and HILIC chromatography) for preparative and analytical services.

Software and Instructional Use

  • Bruker Topspin -Spectra Processing and analysis (in-house).
  • Bruker Dynamics Center -Spectra Processing and Kinetic analysis (in-house).
  • Bruker CMCse -Small Molecule Structure Elucidation (in-house).
  • ACD Labs -Spectral Processing and Analysis (in-house).
  • ChenomxTM -Metabolite Profiling and Quantization (in-house).
  • MVAPCK Statistical Analysis of Spectra (in-house, scripts).

Supplies

The NMR Facility maintains on hand for purchase (or use) the following items:

  • NMR Tubes regular thin walled 7”
  • NMR Tubes (SampleJetTM) 3 mm, 5mm (5’’ and 7’’), individual and boxes of 100 tubes
  • Specialty low volume/high salt NMR tubes
    • Bruker shaped high salt NMR tubes
    • Small volume Shegimi regular and high-salt slotted NMR tubes
  • Deuterated solvents (Chloroform-d, DMSO-d6, Pyridine-d5, MeOH-d4, Acetonitrile-d3)

Time-sensitive services

  • The NMR facility is an open access 24/7 facility for those pre-approved and trained by the facility manager for high end instrumentation use.
  • The facility can accommodate off-campus users with after-hours uses in specific cases for time-sensitive work.

Training

  • Through the engagement of Dr. Brian Tripet, the NMR facility manager, the facility provides high-end training, to enable users with long term research projects to independently operate the various spectrometers. For limited sample users, the facility offers a drop-off/next day result service.
  • Training includes NMR experiment selection. For example, for a 2D 1H-13C HSQC there are 99 parameter sets and 144 pulse sequences. Which is most appropriate for the user is dependent on sample, solvent and concentration. Dr. Tripet streamlines this process while teaching users how to make the right choice.
  • Standard small molecule structure elucidation typically requires a minimum of 6 different types of NMR experiments. Larger molecule structure elucidation requires 10-15 different NMR experiments. Each requires the proper setting of parameters. Dr. Tripet teaches which parameters are most critical to generate data quickly and with high resolution.
  • Training is required for shimming the magnet, tuning and matching the probe, referencing and temperature calibration, pulse calibration for standard and non-standard X nuclei, NUS (Non-Uniform Sampling configuration setup. Undergraduate and graduates are trained on proper setup for integration analysis, and the availability and use of automated scripts for kinetic reaction monitoring. Data analysis software training for Chenomx, Topspin, ACD, Dynamics Center, NMRPipe, Sparky, CMCse and MVAPACK is given. Dr. Tripet has experience to cover all of these topics in depth.

Education

  • The NMR facility also provides in-house lab sections demonstrating NMR hardware/software while conducting lab course experiment/exercises. At present, the NMR facility with Dr. Tripet’s assistance conducts the following sections: 13C NMR analysis of edible oils (CHMY 421), 31P phosphate NMR and kinetics of Alkaline Phosphatase hydrolysis (BCH444), Basic proton and carbon NMR of organic unknowns (CHMY 333), 1D and 2D NMR (CHMY551). Rotational energetics (CHMY374). The NMR Facility is also developing a new graduate-level course in collaboration with Dr. Copié on NMR metabolomics (the course will have both hands on lab experiences and lectures on the theory and best practices for metabolomics research). A pilot version of this course will be offered in Summer or Fall 2019.

Collaborations

  • The NMR facility is open to, and actively encourages collaborations with existing and new users. The facility holds the right to limit such collaborations and refuse such participation if it is deemed the relationship is only for pro-bono services or not in the best interest of the core or its instrumentation.