Equipment

With access to over $26 million in state-of-the-art metabolomics infrastructure, TMIC is able to offer the most comprehensive set of technologies for targeted/quantitative and untargeted metabolomics using NMR, GC-MS, LC-MS, UPLC, HPLC and metabolite imaging. TMIC is able to process 700 samples/week using as little as 20 µL of material. We continuously monitor the status and output quality of all our instruments to guarantee consistent and high quality results. Our scientists and technicians are fully trained in standard operating, maintenance and quality assurance procedures, and have many years experience in handling a wide range of samples and experimental procedures.

See here for the different services we offer and background on the science behind them.

List of Major Equipment in TMIC

NMR

In NMR-based metabolomics, spectral profiling or spectral deconvolution is frequently used to identify and quantify metabolites from 1D 1H NMR spectra. TMIC uses Chenomx Spectral Profiling software to identify and quantify metabolites from 1D 1H NMR spectra. TMIC has also developed a fully automated and quantitative NMR spectral profiling system, BAYESIL. Based on extensive testing with defined mixtures and real biological samples BAYESIL consistently performs with sensitivity and specificity greater than 98% for compound identification in mixtures with up to 60 different compounds. It also determines metabolite concentrations (down to 10 µM) within 10% of the known or expert-measured concentrations. In other words, BAYESIL operates at a level that meets or exceeds the performance of the most highly trained human experts. BAYESIL appears to be the first system that supports fully automated and fully quantitative NMR-based metabolomics. Using NMR-based metabolomics, it is possible to identify and quantify up to 209 metabolites in urine, 55 metabolites in blood, 50 metabolites in CSF, and 45 metabolites in cell or tissue extracts in a single analysis. NMR is ideal for studying amino acids, organic acids, amines, sugars and alcohols.

Find out more about NMR here.

TMIC currently houses the following NMR instrumentation:

500 MHz Varian NMR with 5mm cold probe

600 MHz Varian NMR with 5mm RT probe (2 instruments)

800 MHz Varian NMR with 5mm cold probe

A 700 MHz Varian NMR with 5mm cold probe is also available.

GC-MS

TMIC has developed multiple extraction techniques and uses external calibration-based quantitation for GC-MS analyses. AutoGC software is being developed by TMIC which will semi-automatically identify and quantify metabolites. The NIST08 GC-MS and Golm Metabolome Databases are also used for metabolite identification. With our GC-MS methods, identification and quantification of up to 100 metabolites in urine, 40 metabolites in blood, 60 metabolites in cerebrospinal fluid, and 35 metabolites in cell or tissue extracts is possible. GC-MS is ideal for studying amino acids, biogenic amines, primary amines, organic acids and sugars.

We also offer GC-MS-based lipidomics, using reverse phase HPLC-ELSD to separate lipid classes into 7 groups followed by FAMES and GC-MS analysis to identify and quantify the component fatty acid chains. TMIC-developed Combinatorial Lipid Reconstruction software is used to reconstruct lipid molecules and calculate their concentrations. These lipidomics methods are ideal for identifying/quantifying fatty acids, neutral lipids, phospholipids, and sphingolipids.

Additionally, separation, identification and quantification of small volatile and semi-volatile molecules by two-dimensional GC-MS is offered by TMIC. GCxGC is the gold standard in separating and quantifying exceedingly complex samples of volatile organic compounds, which play a key role in food, flavour and aroma metabolomics, as well as plant and animal communication.

Find out more about GC-MS here.

The following GC-MS and GCxGC instruments are available at TMIC:

Agilent 5695 GC-quad MS with sample changer

GCxGC FID/TOFMS equipped with a liquid nitrogen cryogenic quad jet modulator

GC-MS/MS Bruker SCION TQ GC456 cryo cooling on the PTV, backflush option available

A Agilent 5890 II GC-quad MS with sample changer is also available

HPLC

TMIC offers HPLC-based methods for identifying and quantifying carotenoids, polyphenols, chlorophylls, bile acids and thiols, as well as for lipidomics studies. Our lipidomics workflow includes reverse phase HPLC-ELSD, which separates lipid classes into 7 groups. FAMES and GC-MS analysis are then performed to identify and quantify the component fatty acid chains. Combinatorial Lipid Reconstruction software, developed by TMIC is used to reconstruct lipid molecules and calculate their concentrations. Our lipidomics assays are capable of identifying and quantifying 2000-3000 lipids per sample.

Specialized metabolite assays are also available, using fluorescence-based or UV-based HPLC assays. Specialized metabolite assays include polyphenols, phytochemicals, nucleotides/ nucleosides, vitamins, thiols, bile acids, exotic lipids and extracts.

Find out more about HPLC here.

TMIC houses 6 HPLC/UPLC systems that are dedicated to these analyses:

Beckman System Gold HPLC with ELSD detector

Agilent HPLC system with fluorescence detector

Waters NanoAcquity UPLC system

Waters Acquity UPLC system with UV detector (2 instruments) and Dionex Ultimate 3000 Dual HPLC system with UV detector are also available

LC-MS

TMIC has developed a number of direct injection and LC-MS methods for targeted and untargeted metabolomics. Tandem quadrupole and ion trap instruments are ideal for targeted and quantitative studies. Multiple reaction monitoring (MRM) is used on these instruments to selectively measure specific metabolites and their internal standard, providing absolute quantitation. Our TOF systems provide the high resolution and mass accuracy required for identification of unknowns in untargeted studies. Direct injection analysis has the advantage of speed, and we are currently able to identify and quantify up to 180 different metabolites in a single analysis from blood, urine, saliva, cerebrospinal fluid, tissue and cell extracts, including phospholipids, sphingolipids, lysophospholipids, acylcarnitines, amino acids, and biogenic amines.

TMIC also offers quantitative selective isotope labeling of metabolites (SILOM), which involves chemical modification techniques that are specific to amines, carboxylic acids and hydroxyl groups. This allows for efficient chemo-specific labeling of metabolites with 13C or 12C isotope tags. Spiking known quantities of 13C labelled compounds into biofluids labelled with 12C tags combined with high resolution MS analysis provides identification and quantification of hundreds of metabolites.

In untargeted studies, commercial software (SIMCA by Umetrics) and in-house developed software are used for data processing, including multivariate statistical analysis for identification of changes in metabolite abundance between groups of samples. Untargeted metabolomics studies also utilize UPLC separation followed by Q-TOF MS analysis, or FT-MS analysis, which can yield semi-quantitative data for up to >5000 features per analysis, in as little as 30 minutes.

Find out more about LC-MS here.

The following list of instrumentation includes tandem quadrupole, ion trap, and TOF instruments for targeted and untargeted studies:

ABI QTRAP 4000 MS with Turbo ESI spray probe

ABI QTRAP 4000 MS with Turbo ESI spray probe

Bruker Ion Trap MS with Agilent HPLC system

Agilent 6490 Triple Quadrupole MS

Bruker Q-TOF MS with Agilent HPLC system

Waters Synapt High Defintion Q-TOF MS with ion mobility

MALDI-MS

TMIC is the first facility in Canada to offer MS tissue imaging of endogenous metabolites. This service is available on our high-resolution TOF and FTMS instruments equipped with a MALDI source. Identification and relative abundance information is used to reconstruct colorful ion maps, resulting in a two-dimensional spatial resolution of small molecules (and proteins) in tissue. We can achieve image pixel resolutions of 150 um using FTICR mass spectrometry, and 50 um on the Bruker Ultraflex III MALDI-TOF/TOF.

Find out more about MALDI-MS here.

MALDI-MS analysis is offered on the following instruments:

Bruker TOF/TOF with MALDI-MS for imaging

Applied Biosystems 4800 MALDI TOF/TOF mass spectrometer (3 instruments)

Bruker 11.7 Tesla FT-ICR MS equipped with interchangeable ESI/MALDI source

Bruker Ultraflex III MALDI-TOF/TOF mass spectrometer

ICP-MS

TMIC also offers ICP-MS for quantitative metallomics. We routinely identify and quantify 20-40 metal ions in a wide variety of biological matrices.

A Perkin-Elmer Sciex Elan 6000 Quadrupole ICP-MS with a dual-stage detector is currently used for metal ion studies.

Find out more about ICP-MS here.