Methods and Assays Offered

The MC@CHOP offers a variety of analytical techniques to quantitate stable isotope enrichment (13C, 2H, 15N) and to perform metabolic profiling. Our analytical repertoire includes the following:

  1. Stable Isotope Analysis
    • Stable isotope enrichment (13C, 15N, 2H, 18O) in glucose, amino acids, NH3, urea, tricarboxylic acid cycle intermediates, fatty acids and other intermediates of metabolism

    • 13CO2 enrichment in blood and/or expired air for studies of oxidative metabolism

    • Doubly labelled water (2H2O/H218O ratio) for studies of metabolic rate in a freely moving organism

    • Isotopic enrichment is determined with either:

      1. Liquid chromatography-tandem mass spectrometry

      2. Gas chromatography-mass spectrometry

      3. Isotope ratio mass spectrometry

  2. Measurement – both in vivo and in vitro - of flux through metabolic pathways, such as glycolysis, the tricarboxylic acid cycle, fatty acid oxidation, the urea cycle, protein synthesis/degradation.

  3. Metabolomics profiling including, measurements of amino acids, organic acids, NAD+, NADH, NADP+, NADPH, adenine nucleotides (AMP, ADP, ATP), Pi, triglycerides, free fatty acids, glucose, protein, etc.

  4. Carnitine / Acylcarnitines levels and profiling both in blood and tissue extracts

  5. Measurement of neurotransmitters and neuromodulators levels and profiling

  6. Measurment of D- or L- forms of amino acids and their levels both in physiological fluid and tissue extracts.

  7. Measurment of selected drug and/or drug metabolites as well as determining the impact of a given drug on metabolomics and fluxomics.

  8. Mesurement of signaling compounds such as, cAMP in various tissues.