Metabolomic Core Equipment
The MC@CHOP facility includes a 2,200 square foot laboratory space where equipment is located and analyses are performed. All necessary fume hoods and bio-safety cabinets are part of the core. PC computers, iMacs and MacBooks are available for the staff. The computers are connected to the institutional database and used for data medium, data analyses, and storage.
- Our GC-MS instruments include 3 Hewlett-Packard/Agilent GC-MS, each with an automated sampling system that permits the analysis of up to 99 unattended samples. Each system is controlled by an individual computer/work station with Windows-based proprietary software from HP/Agilent. These instruments are used primarily for the analysis of 15N, 2H or 13C in glucose, amino acids, fatty acids, primary amines and organic acids (particularly tricarboxylic acid cycle intermediates such as citrate, ketoglutarate, succinate, malate).
- Isotope ratio-mass spectrometry for the analysis of 13CO2 in metabolic studies as well as the analysis of 2H2O in doubly-labelled water studies to measure the in vivo metabolic rate of fatty acid oxidation and glycolysis. Instruments include Thermo Finnigan DeltaPLUS continuous flow and Thermo DELTA V Advantage IRMS, equipped with an autosampler that permits unattended analysis of 96 samples. Sensitivity of the latter instrument (atom % excess) characteristically exceeds 10-5.
- Two LC-MS triple stage quadrupole systems (a VG Quattro II LC-MS unit and an Agilent 6400 LC-triple stage quadrupole system). The latter is used to perform metabolomic research, or the profiling in a single sample of > 100 individual metabolites to create a metabolic "fingerprint."
- HPLC instruments include two Agilent-1260 Infinity HPLC systems equipped with UV and fluorescent detectors, and are used to measure metabolite levels as an integral part of stable isotope analysis and determination of flux rates.
- Enspire 2300 multilabeled Reader for monitoring of enzyme assays, and other spectrophotometers used for enzymatic assays.
- Systems to monitor oxygen consumption in isolated mitochondria or cells, whole organ (e.g., liver, kidney, heart), perfusion systems as well as all necessary centrifuges, balances, pH meters, metabolic incubators, cold room, refrigerators and freezers for sample preparation and storage.