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 Nuhnb, 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.