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Equipment/Services

Metabolomics and Bioenergetics Core

VAI’s Metabolomics and Bioenergentics Core offers a range of comprehensive metabolism and mass spectrometry services along with consulting and training for VAI investigators.

Thermo Fisher Orbitrap ID-X UHPLC/MS

Primary use: Untargeted and semi-untargeted metabolite profiling and stable isotope labeling

Capabilities

  • High-resolution/accurate mass data acquisition
  • Up to four chromatographic separations per sample to maximize metabolite coverage
  • MS2 fragmentation for compound identification and library matching
  • MSn fragmentation for structural elucidation

Specifications

  • Up to 500k resolution (FWHM)
  • < 2ppm mass accuracy
  • Electrospray ionization
  • Dual Vanquish UHPLC
  • Data dependent acquisition
  • CID and HCD fragmentation

Agilent 1290 UHPLC/ 6470 QQQ MS

Primary use: Targeted polar metabolite profiling and custom targeted method development

Capabilities

  • Ion-paired negative mode profiling of ~250 polar metabolites, including amino acids, nucleotides, carboxylates and sugar phosphates
  • dMRM mode limits transition monitoring only to retention windows of interest to maximize compounds detected per run
  • Highly reproducible ion-paired chromatography
  • High sensitivity and specificity of targeted compounds
  • Second UHPLC system for targeted method development of compounds not amenable to ion-pairing.

Specifications

  • Sub-femtogram detection limits (compound dependent)
  • Electrospray ionization

Agilent 7890 GC/5977 MSD

Primary use: Targeted metabolite profiling of central carbon metabolism and custom targeted method development

Capabilities

  • Profiling, quantitation, and stable isotope labeling of most amino acids and TCA cycle intermediates
  • In-source compound fragmentation for positive ID

Specifications

  • Sub-femtogram detection limits (compound dependent)
  • Electron-impact ionization

Seahorse XFe96

Primary use: Measure OCR and ECAR of live cells in a 96-well plate format. OCR and ECAR rates are key indicators of mitochondrial respiration and glycolysis and these measurements provide a systems-level view of cellular metabolic function in cultured cells and ex vivo samples

Capabilities

  • Four-port injection system and automated mixing allows real-time detection of responses to substrates, inhibitors and other compounds. Temperature is maintained at 37oC
  • A 96-well plate allows multiple conditions to be run at once. The system is optimized for compound screening and dose-response studies.
  • As few as 5,000 cells per well may be analyzed at high sensitivity (depending on cell type)
  • System is compatible with many different sample types

Specifications

  • 96 assay wells
  • 2µL microchamber volume