DataEntryID 54
    General information
      Manuscript title: Quantification of intracellular metabolites in Escherichia coli K12 using liquid chromatographic-electrospray ionization tandem mass spectrometric techniques.
      PubMed ID: http://www.ncbi.nlm.nih.gov/pubmed/11488613
      Journal: Analytical Biochemistry
      Year: 2001
      Authors: Arne Buchholz, Ralf Takors, and Christian Wandrey
      Affiliations: Institute of Biotechnology, Research Center Juelich, 52425 Juelich, Germany
      Keywords: Escherichia coli, intracellular metabolites, ESI, extraction, LC-MS, quantification
      Full text article: no file uploaded
      Project name: not specified

    Experiment description
      Organism: Escherichia coli
      Strain: K-12 DSM
      Data type: metabolites at steady-state
      Data units: mM
      Execution date: not specified

    Experimental details
      Temperature (°C): not specified
      pH: not specified
      Carbon source: glucose
      Culture mode: chemostat
      Process condition: aerobic
      Dilution rate (h⁻¹): 0.125
      Working volume: 0.4 L
      Biomass concentration (g/L): not specified
      Medium composition: Minimal medium: 30 g/L glucose, 0.2 g/L NH4Cl, 2.0 g/L (NH4)2SO4, 3.25 g/L KH2PO4, 2.5 g/L K2HPO4, 1.5 g/L NaH2PO4, 0.5 g/L MgSO4; trace substances: 1.0E-2 g/L CaCl2, 5.0E-4 g/L ZnSO4, 2.5E-4 CuCl2, 2.5E-3 g/L MnSO4, 1.75E-3 g/L CoCl2, 1.25E-4 g/L H3BO3, 2.5E-3 g/L AlCl3, 5.0E-4 g/L Na2MoO4, 1.0E-2 g/L FeSO4.
      General protocol information: Sampling Method: Samples are taken after the cultures have remained constant at a cell density of 10.0 g/L for at least three residence times, thus ensuring steady-state conditions.; Quenching: Sampling was performed using precooled syringes containing 15 ml of the quenching fluid, 60%, v/v, aqueous methanol containing 70 mM Hepes (2-[4-(2-hydroxyethyl)- 1-piperazinyl]ethanesulfonic acid).; Extraction list: methanol-water; Analysis list: LC-ESI-MS;
      Methods description: Identification and Quantification of Metabolites - compounds were identified in cell extracts using retention time, m/z, specific fragmentation patterns, and spiking with known compounds. Quantification was accomplished via the [M-H]2 ion by applying the standard addition method [1]. A standard solution containing all analytes at a known concentration was prepared. By spiking cell extracts with increasing amounts of this standard solution, linear regression plots of peak area versus concentration were obtained. At least three data points were averaged per concentration in a concentration range of 0.005–1.5 mM. Limits of detection (LOD) were determined by taking three times the standard error of the estimate for the regression of these linear regression plots.

------------References------------
[1] Skoog, D. A., and Leary, J. J. (1992) Principles of Instrumental Analysis, Saunders, New York.
      Data file: http://kimosys.org/repository/54/download?parameter=1138; 
      Alternative formats: http://kimosys.org/repository/54/attached_files/130/download; http://kimosys.org/repository/54/attached_files/31/download; 

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      Entered by: Administrator KiMoSys
      Created: 2013-05-29 15:18:00 UTC
      Updated: 2020-04-24 16:10:32 UTC
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