Targeted Metabolomics

This service is provided by the Biomarkers Core Laboratory.

Overview

Targeted metabolomics is the hypothesis-driven, absolute quantitative assessment of a subset of metabolites often linked to a metabolic pathway (glycolysis, TCA cycle).

Biomarkers Core Laboratory (BCL) offers several Liquid Chromatography-Mass Spectrometry (LC-MS/MS) based targeted metabolomics assays to support preclinical, clinical and translational research. Core services also include the method development and validation of novel mass spectrometry - based targeted quantitative assays for the metabolites of your interest.

Our targeted metabolomics assays involve a reverse phase or HILIC based chromatographic separation followed by either selective or multiple reaction monitoring (SRM or MRM) in positive or negative electrospray ionization mode, depending on the nature of the analyte(s). The quantitative approach is based on the use of stable isotope labeled internal standards (13C, 2H) and external reference standards. Core customizes the sample preparation protocols (protein precipitation, Liquid-Liquid extraction and Solid Phase extraction) based on the physicochemical properties of the compound, their abundance, and the sample type among other factors.

Metabolomics Assays

BCL offers several targeted LC-MS /MS based assays for individual biomarkers, as well as panels targeting specific classes of metabolites. These assays are validated against a wide range of sample types including body fluids (plasma, serum, saliva, urine), cells, and various tissue types from human subjects and research animals.

The major metabolomics panels and the analytes include:

Biocrates MxP® Quant 500 metabolites Kit: BCL offers a comprehensive targeted metabolite quantitation using the MxP® Quant 500 kit (BIOCRATES Life Sciences AG, Innsbruck, Austria) with a coverage of up to 630 metabolites from 26 analyte classes. The Q500 kit is validated for use with both human and animal plasma, serum, tissue, feces, and cells. Metabolite coverage includes aminoacids and aminoacids related (#50), bile acids (#14), acylcarnitines (#40), fatty acids (#12), phosphatidylcholines (76), and triglycerides (242). The complete list can be found here.

Bile acids: CA, DCA, UDCA, HDCA, LCA, TLCA, TCDCA, GCA, TDCA, GDCA, TCA, GCDCA, GUDCA, TUDCA, (α+Ɯ)-MCA, β-MCA, and T(α+β)-MCA.

Acylcarnitines: Carnitine (C0), Acetylcarnitine (C2), Propionylcarnitine (C3), Butyrylcarnitine (C4), Isovalerylcarnitine (C5), Octanoylcarnitine (C8),  Myristoylcarnitine (C14), and Palmitoylcarnitine (C16)

Aminoacids:  Histidine, Arginine, Asparagine, Glutamine, Serine, Aspartic acid, Citrulline, Glutamic acid, Threonine, Alanine, Proline, Cystine, Lysine Tyrosine, Leucine, Isoleucine, Valine, Methionine, and Glycine

Catecholamine metabolites: Dopamine, Epinephrine, Norepinephrine, DOPAC, L-DOPA, DHPG, Methoxy tyramine, homovanillic acid 

Tryptophan Metabolites: Tryptophan, Kynurenine, Serotonin, Kyurenic acid, 3-Hydroxy Kyurenine, 3-Hydroxyanthranilic acid, 5-Hydroxyindole-3-acetic acid,  Quinolinic acid, Anthranilic acid

Polyamines: Putrescin, Spermine, and Spermidine

Free Fatty acids: Lauric acid (C12:0), Tridecanoic Acid (C13:0), Myristic acid (C14:0), Palmitic acid (C16:0), Stearic acid (C18:0), Oleic acid (C18:1), Linoleic acid (C18:2), alpha Linolenic acid (C18:3), Arachidic acid (C20:0), Arachidonic acid (C20:4), Eicosapentaenoic acid (C20:5), Arachidonic Acid (C22:4), , Docosapentaenoic acid (C22:5), and Docosahexaenoic acid (C22:6).

Tricarboxylic acids (TCA) :Citric acid, Isocitric acid, Fumaric acid, Succinic acid, Malic acid, Oxaloacetic acid, α-Ketoglutaric acid, Lactic acid, Aspartic acid, 2-Hydroxyglutaric acid, and Glutamic acid.

Short-chain fatty acids (SCFA):Acetic acid, Propionic acid, Butyric acid, Isobutyric acid, Valeric acid, Isovaleric acid, Caproic acid, Isocaproic acid

Steroids: Estradiol, Progesterone, Testosterone, Estrone

Vitamin D metabolites: 25-hydroxyvitamin D2, 25-hydroxyvitamin D3

Note: In a few instances, metabolites included in the same class run as separate assays due to their varying dynamic range or chromatographic properties.

Instrumentation

Waters Xevo TQ-S

Xevo TQ-S MS from Waters Corporation features a StepWave ion guide that delivers the highest levels of sensitivity and robustness. It also has an innovative ScanWave™ collision cell technology to provide enhanced spectral LC-MS/MS data acquisition capabilities. At BCL, the TQ-S MS is integrated with Waters® Acquity UHPLC system for the sensitive detection of steroids, contraceptive drugs, and neurotransmitters among others. Integration of TQ-S with Waters® nanoacquity UPLC through the ionKey source allow us to employ the 150uM I. D. plug and play iKey separation device enabling highly efficient, sensitive microflow separations for the low abundant protein turnover applications.

 

Waters Xevo TQ-Absolute

The Waters Xevo™TQ absolute mass spectrometerwith the Binary Acquity Premier LC system has a sensitivity of 1pg on the column, dual mode ESi/APCi source, and capable of switching between MS and MS/MS in 3msec in a single injection. These features offer enhanced capabilities to measure low abundant metabolites often present at sub-nanomolar concentrations. TQ absolute MS is specifically designed for better performance at negative ionization mode, delivering up to 6 to 15X more sensitivity than other triple quadrupole instruments, which is ideal for the quantitation of most primary metabolites. Binary Acquity premier LC system is unique with its MaxPeak High-Performance Surfaces (HPS) Technology, designed specifically to improve the peak shape and reproducibility of the most challenging metal-sensitive compounds

 

Hours

The BCL is open Monday – Friday, 9am to 5pm. Samples may be dropped off during these hours and dry ice must be used during transportation.

Eligibility and Requirements

Targeted metabolomics services are available to all researchers at Columbia University and at other academic institutions.   

Investigators who are not affiliated with Columbia University will need to complete a service agreement that must be signed by the legal or purchasing departments of both parties. This process takes approximately four weeks to finalize. The requester's legal or purchasing department’s service agreement template is typically used, but one can be provided by Columbia if necessary. For more information on service agreements and pricing for non-CU initiated studies, please email the BCL director, Dr. Renu Nandakumar.

The BCL is not Clinical Laboratory Improvement Amendments (CLIA)-certified. The data generated from BCL is to be used for research purposes only and not for clinical and diagnostic applications.

Submitting Requests

A new protocol submission request must be initiated in iLab for every new project. Samples will not be accepted without an active service request. Additional request details forms must be submitted if more assays or samples need to be added to an existing protocol. Please contact us for the instructions on how to register and submit a service request through iLab.

Sample Submission

Please review the BCL’s User Policy and General Sample Submission Guidelines prior to submitting the samples and complete the downloadable sample list template to be delivered along with your samples.

When to Request This Service

We strongly encourage investigators to contact BCL director, Dr. Renu Nandakumar, prior to submitting a new service request. This will facilitate a discussion on the project, scope, feasibility, appropriate analytical strategy and other sample requirements. Please set up a consultation to initiate new metabolomics projects involving LCMS method development.

Cost

A complete list of the mass spectrometry-based assays and associated cost is available on our iLab site. If you do not find the assay that you are interested in, please contact us and we will work with you to explore your options.

Cite it, Submit it, Share it!

If your research has benefited from one or more Irving Institute resources, please remember to:

• Cite our CTSA grant, UL1 TR001873, in any relevant publications, abstracts, chapters, and/or posters.
• Submit your publications to PubMed Central (PMC) for compliance with the NIH Public Access Policy.
• Share your research updates with us by sending an email to: irving_institute@cumc.columbia.edu