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

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-	=== Column ===	+	=== Column ===
-	if used	+
-	* Type:	+
-	* Temperature:	+
		+	if used
		+	*Type: silica column LiChroCart 125-4 with Si 60 (5µm) (merck)
	=== Mass spectrometer ===		=== Mass spectrometer ===

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Revision as of 11:53, 2 September 2008

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Contents 1 Sample preparation 2 Instrumentation and method 2.1 Pump 2.2 Column 2.3 Mass spectrometer 3 Data analysis and quantification 3.1 Data handling 3.2 Isotope correction 3.3 Calibration and quantification 4 Method validation 4.1 Accuracy 4.2 Precision 4.3 Limit of detection 4.4 Recovery 5 Sample data 5.1 Mass spectra 5.2 Chromatogram 6 Reference

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

if used

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

Mass spectrometer

• Type:
• Ionization mode:
• Ionization voltage:
• Source temperature(s):
• Collision gas:
• Collision gas pressure:
• MS/MS-conditions:
  • multiple reaction monitoring table

Analyte	Precursor [m/z]	Precursor [m/z]	Collision energy [eV]
species 1	m/z 1	m/z 2	x
species 2	m/z 1	m/z 2	x

Data analysis and quantification

Data handling

• chromatogram, spectrum and peak processing

Isotope correction

• if necessary

Calibration and quantification

• calibration type: matrix or external calibration by standard addition to sample matrix
• species used for calibration

Species	Material 1	Material 2
Species 1	range pmol	range pmol
Species 1	range pmol	range pmol

Method validation

Accuracy

Precision

Limit of detection

Recovery

Sample data

Mass spectra

Chromatogram

Reference

• PubMed
• electronic journal

Please add categories as below for:

• analytical techniques applied
• lipid classes analyzed (see lipid class categories)