Lipid and Metabolite Mass Spectrometry Facility
The Center for Human Nutrition lipid mass spectrometry lab is in the Green Science Building (L4.308) and occupies 2,000 square feet of newly renovated, state-of-the-art laboratory space housing over 16 mass spectrometers. This is a newly constructed open-concept lab with a modular design that allows equipment to relocated with minimal effort or disruption based on current needs.
Other than two chemical fume hoods and a sink, there are no fixed structures in the lab. Nitrogen and ultra-pure air are delivered from an external 6,000-gallon liquid nitrogen dewer and twin (redundant) Atlas Copco SF22+ scroll compressors with CD35+ dryers. All gasses are delivered through a fully stainless steel, medical grade piping network.
The lab is temperature controlled through a combination of house HVAC and four in-room water chilled CRAC units. Mechanical services (gas, electrical, and vent snorkels) are delivered primarily through ceiling mounted quick connect gas fittings, twist-lock power cords, and snorkels with additional electrical service along the walls.
There is currently 64 feet of mobile benches (AT Villa Ergo) for sample preparation, instrument maintenance, and other activities related to lipid mass spectrometry.
- SCIEX 4000 QTRAP Triple Quadrupole Mass Spectrometer
Shimadzu LC-20xr HPLC with ternary gradient capabilities (3 pumps), autosampler, column oven and degasser.
Primary use: Targeted analysis of lipid classes, method development, unique exploratory methods.
- Two SCIEX API 5000 Triple Quadrupole Mass Spectrometers
Shimadzu LC-20xr HPLC with ternary gradient capabilities (3 pumps), autosampler, column oven and degasser.
Primary use: targeted, quantitative analysis of sterols, oxysterols, bile acids, steroids, eicosanoids, and sphingolipids.
- SCIEX TripleTOF© 6600 Quadrupole time-of-flight Mass Spectrometer
LEAP Infuse PAL automated sample infusion platform.
Primary use: High-throughput untargeted lipidomic profiling using MS/MSALL workflows.
- SCIEX QTRAP© 6500 Triple Quadrupole/Ion Trap Mass Spectrometer
Shimadzu LC-30 Nexera HPLC with ternary gradient capabilities (3 pumps), autosampler, column oven and degasser with 4th pump for post-column addition of reagents.
Primary use: Semi-targeted lipidomic profiling using scan and MRM with normal-phase chromatography.
**Shared resource with the Manglesdorf/Kliewer laboratory in Pharmacology
- SCIEX QTRAP© 4500 Triple Quadrupole/Ion Trap Mass Spectrometer
Shimadzu LC-20AD HPLC with ternary gradient capabilities (three pumps), autosampler, column oven and degasser with 4th pump for post-column addition of reagents.
Primary use: Semi-targeted lipidomic profiling using scan and MRM with normal-phase chromatography.
**Shared resource with Tambar/Kohler laboratory in Biochemistry
- Agilent 7890A/5975c
Gas chromatograph quadrupole mass spectrometer with EI and PCI/ECNI capabilities.
Primary use: analysis of fatty acids when mass specificity is needed including stable isotope labeling experiments. Also used when increased detectability is required for specific fatty acids or non-esterified fatty acids.
- Agilent 7890C/5975c
Gas chromatograph quadrupole mass spectrometer.
Primary use: Analysis of metabolites and metabolic tracing.
**Shawn Burgess Lab
- Agilent 6890N/5973
Gas chromatograph quadrupole mass spectrometer with EI and PCI/ECNI capabilities.
Primary use: Analysis of sterols by and other lipids where increased chromatographic separation is needed not attainable by liquid chromatography methods.
- Thermo Scientific™ Orbitrap Fusion™ Lumos™ Tribrid™ 1M Mass Spectrometer
Shimadzu LC-30 Nexera HPLC with ternary gradient capabilities (3 pumps), autosampler, column oven and degasser.
Primary use: Lipid flux analysis using deuterated water as a substrate. The resolution of this mass spectrometer allows for the mass resolution of the 2H signal from the endogenous 13C signal. Data can be collected via the HPLC interface or by direct infusion.
**Matthew Mitsche Lab
- Thermo Scientific™ Q Exactive™ GC Orbitrap™ Mass Spectrometer
Thermo Scientific Trace 1310 GC system with EI/PCI/ECNI capabilities
Primary use: Denovo lipogenesis analysis (cholesterol and fatty acids) using deuterated water as the substrate. The resolution of this mass spectrometer allows for the mass resolution of the 2H signal from the endogenous 13C signal for lipids below ~ 300 Da. This platform complements the Fursion Lumos LC-MS for lipid and metabolite flux analysis for analytes better suited to GC analysis.
- SCIEX 3200 QTRAP Triple Quadrupole Mass Spectrometer
Shimadzu LC-20xr HPLC with ternary gradient capabilities (three pumps), autosampler, column oven and degasser.
Primary use: Targeted analysis of lipid classes, method development, unique exploratory methods.
**Shawn Burgess Lab
- Agilent 6980 Gas Chromatograph
Gas chromatograph with flame ionization detector.
Primary use: general fatty acid analysis and to confirm fatty acid measurements made with other analytical methods.
- Thermo Ultimate 3000 UHPLC
Contains Corona Ultra RS detector.
Primary use: Lipid class quantitation.
Sample Preparation:
- Hamilton Starlet liquid handling robot
- Biotage 96-well positive pressure SPE manifold
- Biotage 96-well nitrogen dryer
- (3) Thermo-Pierce 27-position nitrogen dryers
- (2) Labconco 50-position RapidVap Vertex nitrogen dryers
- Omni BeadRuptor bead beater/homogenizer
- (2) Benchtop centrifuges
- (2) Micro centrifuges
Sample Storage:
- (2) -80 °C freezers
- (2) -20 °C solvent-safe freezers
- Lab refrigerator
Air and Nitrogen Supply:
- Parker LCMS 5000 ultra-pure air and nitrogen generator to supply the API 5000 instruments
- 6000-gallon liquid nitrogen dewar to supply nitrogen to the L4.308 CHN mass spectrometry laboratory.
- (2) Atlas Copco SF22+ scroll compressors each with CD35+ air dryers and particle filters to generate "zero" air to L4.308 CHN mass spectrometry laboratory.