Ceramides - stratum corneum, free extractable - LC-MS - Farwanah et al.

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Contents

Sample preparation

Material:  Palmoplantar stratum corneum scrapings from healthy volunteers

Extraction:  Chloroform–Methanol (1:1, v/v)

Instrumentation and method

Pump

  • Type: Spectra System P 4000  ( Thermo separations)
  • Mode: Gradient
  • Solvent(s): A: chloroform; B: chloroform, n- propanol, acetic acid (80:20:2) 
  • Gradient:
Time [min] Flow [ml/min]  % Solvent A  % Solvent B
0 1 100 0
15 1 0 100

Column

  • Type: silica column LiChroCart 125-4 with Si 60 (5µm) (merck)

Mass spectrometer

  • Type: Finnigan LCQ classic (ion trap mass spectrometer)
  • Ionization mode: APCI, positive full scan mode
  • Source temperature(s):  APCI heater:  set to 500°C,  heated capillary: 150°C


Reference

Hany Farwanah, Peter Nuhn, Reinhard Neubert, Klaus Raith. Normal-phase liquid chromatographic separation of stratum corneum ceramides with detection by evaporative light scattering and atmospheric pressure chemical ionization mass spectrometry. Analytica Chimica Acta 492 (2003) 233-239.

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