The services provided by the Yale DRCClinical Metabolism Core include the extraction, purification, derivatization, and instrumental analysis of concentrations and stable-isotopic enrichment of metabolites from tissues and plasma. The methods have been scaled to work with human and rat biopsy tissues of ~25 mg, and plasma volumes of ~5 to 25 µl. Purification techniques include ion exchange chromatography, dialysis, centrifugation or other appropriate techniques. Derivatization is determined by the chemical structure of the metabolite, the isotopic analysis desired, and the instrumental method employed. Specifically, the Core provides the following services:
- Preparation of extracts of tissue and plasma samples for NMR, LC-MS/MS, GC-MS, and COBAS analysis.
- Measurement of total concentrations of metabolites from plasma and urine originated from rat and mouse studies of diabetes and related metabolic diseases. These metabolic panels include Chem 7, liver function tests, lipid profiles, and divalent ions.
- Measurement of the plasma and tissue concentration profiles of lipid metabolites associated with the development and progression of diabetes and related metabolic diseases.
- Measurement of isotopic (e.g., 2H, 13C, 15N, and 18O) enrichment and concentrations of cytoplasmic and mitochondrial intermediary metabolites of glycolysis and the tricarboxylic acid cycle for determining energy production and mitochondrial function.
- Measurement of isotopic (e.g., 2H, 13C, 15N, and 18O) enrichment and concentrations of over 140 intermediary metabolites by GC-MS, LC/MS/MS, and NMR for the calculation of turnover of carbohydrates, lipids, and proteins.