Equipment

The most comprehensive set of technologies for targeted quantitative and untargeted metabolomics.

Major Equipment
NMR
GC-MS
HPLC
LC-MS
ICP-MS
CE-MS

Access to over $30 million in state-of-the-art metabolomics infrastructure!

Targeted/quantitative and untargeted metabolomics using NMR, CE-MS, GCxGC MS, GC-MS, LC-MS, UPLC, HPLC, and metabolite imaging.

TMIC is able to process 700 samples/week using as little as 20 µL of sample. 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 of experience in handling a wide range of samples and experimental procedures.

 

Major Equipment

Nuclear Magnetic Resonance (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, MagMet. Based on extensive testing with defined mixtures and real biological samples MagMet 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, MagMet operates at a level that meets or exceeds the performance of the most highly trained human experts. MagMet 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.

TMIC currently houses the following NMR instrumentation:

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

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

GC-MS and Multidimensional GCXGC 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. This platform is ideal for the analysis of volatile and semi-volatile compounds, as well as those compounds which can be derivatized with a standard methoximation / silylation chemistry which permits us to see sugars, sterols, and larger molecules. We also have a wide range of sample introduction methods available that can be used with GC×GC-TOFMS in addition to standard liquid injections. These include capabilities for large-volume injection, automated headspace, solid-phase microextraction (SPME), dynamic headspace, and thermal desorption from sorbent tubes. These allow us to easily tackle samples such as volatiles from soil, water, plants, food and beverages, and breath, in addition to more conventional biosamples (urine, plasma, etc).

The Following GC-MS And GCxGC Instruments Are Available At TMIC:
–An Agilent 5890 II GC-quad MS with sample changer is also available
 
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

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.

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

Also Available:
-Waters Acquity UPLC system with UV detector (2 instruments)
-Dionex Ultimate 3000 Dual HPLC system with UV detector

Beckman System Gold HPLC with ELSD detector
Agilent HPLC system with fluorescence detector
Waters NanoAcquity UPLC system

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.

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
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

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.

Perkin-Elmer Sciex Elan 6000 Quadrupole ICP-MS

CE-MS

Capillary electrophoresis (CE) offers versatile separation modes for global metabolomic analysis, and is especially well-suited to the analysis of polar/ionic metabolites from volume-limited (< 5uL) or mass-limited (< 5 mg dried weight) specimens. This is a major benefit to metabolomic studies involving precious bio-banked samples (3.2 mm dried blood spot punch), human tissue biopsies (muscle), or animal (mouse) models. Our CE-MS group also specializes in multisegment injection CE-MS for high throughput biomarker discovery which has proven to be particularly useful in large-scale studies involving thousands of samples.

We request that any publications, posters, or oral presentations resulting from work performed by The Metabolomics Innovation Centre acknowledge TMIC. This allows the Centre to report information such as impact on Canadian researchers to our funding agencies. Clients should also keep us apprised of any publications, posters, or oral presentations arising from work performed at TMIC.

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