Ceramide - ESI-MS/MS - Liebisch et al.

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* G.Liebisch, W. Drobnik, M. Reil, B. Trümbach, R. Arnecke, B. Olgemöller, A. Roscher, G. Schmitz ''Journal of Lipid Research'' '''1999''', ''40'', 1539-1546.
* G.Liebisch, W. Drobnik, M. Reil, B. Trümbach, R. Arnecke, B. Olgemöller, A. Roscher, G. Schmitz ''Journal of Lipid Research'' '''1999''', ''40'', 1539-1546.
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Institut für Klinische Chemie und Laboratoriumsmedizin; Klinikum der Universität Regensburg; Franz-Josef-Strauss-Allee 11; D-93042 Regensburg; Germany.
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[http://www.ncbi.nlm.nih.gov/sites/entrez PMID=10428992]
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Fax: 49-941-944-6202; e-mail: gerd.schmitz@klinik.uni-regensburg.de
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[[Category:Flow injection analysis]]
[[Category:Flow injection analysis]]
[[Category:Triple quadrupole]]
[[Category:Triple quadrupole]]

Revision as of 19:12, 27 April 2008

Contents

Sample preparation

  • lipid extraction according to Bligh and Dyer
  • internal standards are added prior lipid extraction
  • to avoid the loss of lipids, extraction was performed in glassware
  • samples are dissolved in methanol containing 5 mM ammonium formate/chloroform (3/1 = v/v)
Material Material used Internal Standard(s) Internal Standard(s) added
Cultured cells 100µg protein Cer 14:0, Cer 17:0 x ng
Human plasma 20µl Cer 14:0, Cer 17:0 x ng


Instrumentation and method

Pump

  • Type: binary high pressure gradient (Agilent 1100)
  • Mode: isocratic flow gradient
  • Solvent(s): 5 mM ammonium formate/chloroform (3/1 = v/v)
  • Gradient:
Time [min] Flow [ml/min]  % Solvent A  % Solvent B
0 0.05 100 0
0.1 0.03 100 0
1.1 0.2 100 0
1.3 0.05 100 0

Autosampler

  • Type: CTC Pal
  • Injection volume: 20µl
  • Wash solvent: methanol/chloroform = 1/1

Mass spectrometer

  • Type: Triple quadrupole (Micromass, Quattro Ultima)
  • Ionization mode: ESI positive
  • Ionization voltage:
  • Source temperature:
  • Collision gas: Argon
  • Collision gas pressure:
  • MS/MS-conditions:
    • precursor ion scan of m/z 264.2
    • multiple reaction monitoring table
Analyte Precursor [m/z] Precursor [m/z] Collision energy [eV]
Cer 14:0 534 xx


Data analysis and quantification

Data handling

Isotope correction

Calibration and quantification

Method Validation

Accuracy

Precision

LOD

LOQ

Recovery

Reference

  • G.Liebisch, W. Drobnik, M. Reil, B. Trümbach, R. Arnecke, B. Olgemöller, A. Roscher, G. Schmitz Journal of Lipid Research 1999, 40, 1539-1546.

PMID=10428992

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