Offline nanoESI Phospholipid Profiling - LC-MS/MS

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Sample preparation

  • liquid-liquid extraction according to Bligh & Dyer
  • sample sovlent: Lipid extracts are diluted 1:100 in acetonitrile/2-propanol 5:2
Material Material used Internal Standard(s) Internal Standard(s) added
Lipid extracts 10µl 24:0PC, 24:0PE, 24:0PS 500pmol each

Instrumentation and method

Mass spectrometer

  • Type: Thermo TSQ Quantum Ultra
  • Ionization mode: nanoESI
  • Ionization voltage: 1500V
  • Capillary temperature: 220°C
  • Capillary voltage: 35V
  • Collision gas: Argon
  • Collision gas pressure: 0.7mTorr
  • MS/MS-conditions:
    • scan range: m/z 400-900
    • resolution: 0.7 peak width FWHH
    • scan modes
Analyte Precursor of [m/z] Neutral loss [MW] Collision energy [eV]
PC 184 35
PE 141 25
PS 185 25

Data analysis and quantification

Data handling

  • Spectra are smoothed and peak lists exported to Excel.

Calibration and quantification

  • calibration type: relative quantification
  • The peak list generated from every headgroup scan is compared with a list of theoretical m/z values for every phospholipid class. Following peak identification the internal standard is set to 100% and all other peaks are expressed as % relative to internal standards.

Method validation


Limit of detection

Sample data

Mass spectra



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