Ceramides - stratum corneum, free extractable - LC-MS - Farwanah et al.
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- | + | == Sample preparation == | |
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+ | Material: Palmoplantar stratum corneum scrapings from healthy volunteers | ||
- | + | Extraction: Chloroform–Methanol (1:1, v/v) | |
- | + | == Instrumentation and method == | |
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- | + | === Pump === | |
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- | + | *Type: Spectra System P 4000 ( Thermo separations) | |
+ | *Mode: Gradient | ||
+ | *Solvent(s): A: chloroform; B: chloroform, n- propanol, acetic acid (80:20:2) | ||
+ | *Gradient: | ||
- | + | {| class="wikitable" style="text-align: center" border="1" | |
- | + | |- | |
- | + | ! Time [min] | |
- | + | ! Flow [ml/min] | |
- | + | ! % Solvent A | |
- | + | ! % Solvent B | |
- | {| | + | |
- | ! Time [min] | + | |
|- | |- | ||
! 0 | ! 0 | ||
- | | | + | | 1 |
+ | | 100 | ||
+ | | 0 | ||
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- | ! | + | ! 15 |
- | | | + | | 1 |
- | + | | 0 | |
- | + | | 100 | |
- | | 0 | + | |
|} | |} | ||
- | === | + | === Column === |
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- | + | *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<br> | ||
- | + | <br> | |
- | == Reference == | + | == Reference == |
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+ | 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. | ||
- | + | [[Category:APCI]] [[Category:Single_quadrupole]] [[Category:Ceramides_(SP02)]] [[Category:LC-MS]] | |
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- | [[Category: | + |
Current revision
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.