Ceramides - stratum corneum, free extractable - LC-MS - Farwanah et al.
From LipidomicsWiki
(Difference between revisions)
Line 1: | Line 1: | ||
How to use: | How to use: | ||
* Click 'Edit' and copy the source code to clipboard | * Click 'Edit' and copy the source code to clipboard | ||
- | * Create | + | * Create new page/article and paste the source code into your new article. |
- | * | + | * Modify as needed. |
- | |||
- | == | + | == Sample preparation == |
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | * liquid-liquid extraction, SPE .... | |
+ | * sample sovlent: | ||
+ | {| class="wikitable" style="text-align:center" | ||
+ | ! Material !! Material used !! Internal Standard(s) !! Internal Standard(s) added | ||
+ | |- | ||
+ | ! Cultured cells | ||
+ | | 100µg protein || Cer 14:0, Cer 17:0 || 50ng each | ||
+ | |} | ||
- | == | + | == Instrumentation and method == |
- | == | + | === Pump === |
+ | * Type: | ||
+ | * Mode: isocratic or gradient | ||
+ | * Solvent(s): | ||
+ | * Gradient: | ||
+ | {| border="1" class="wikitable" style="text-align:center;" | ||
+ | ! Time [min]!! Flow [ml/min] !! % Solvent A !! % Solvent B | ||
+ | |- | ||
+ | ! 0 | ||
+ | | 0.2 || 100 ||0 | ||
+ | |- | ||
+ | ! 1.0 | ||
+ | | 0.2 || 50||50 | ||
+ | |- | ||
+ | ! 1.5 | ||
+ | | 0.2 || 100 ||0 | ||
+ | |} | ||
+ | === Autosampler === | ||
+ | * Type: | ||
+ | * Injection volume: | ||
+ | * Wash solvent: | ||
- | == | + | === Mass spectrometer === |
- | == | + | * Type: |
+ | * Ionization mode: | ||
+ | * Ionization voltage: | ||
+ | * Source temperature: | ||
+ | * Collision gas: Argon | ||
+ | * Collision gas pressure: | ||
+ | * MS/MS-conditions: | ||
+ | ** precursor ion scan of m/z 264.2 | ||
+ | ** multiple reaction monitoring table of species observed frequently | ||
+ | {| border="1" class="wikitable" style="text-align:center" | ||
+ | ! 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 | ||
+ | * both [M+H]+ and [M+H-H2O]+ are used | ||
+ | * species used for calibration | ||
+ | {| class="wikitable" style="text-align:center" | ||
+ | ! Species!! Cultured cells !! Human plasma | ||
+ | |- | ||
+ | ! Cer 16:0 | ||
+ | | 0-35pmol || 0-50pmol | ||
+ | |- | ||
+ | ! Cer 18:0 | ||
+ | | 0-35pmol || 0-50pmol | ||
+ | |- | ||
+ | ! Cer 20:0 | ||
+ | | 0-35pmol || 0-50pmol | ||
+ | |- | ||
+ | ! Cer 24:1 | ||
+ | | 0-35pmol || 0-50pmol | ||
+ | |- | ||
+ | ! Cer 24:0 | ||
+ | | 0-35pmol || 0-50pmol | ||
+ | |} | ||
- | |||
+ | == Method validation == | ||
- | === | + | === Accuracy === |
+ | === Precision === | ||
- | === | + | === LOD === |
+ | === LOQ === | ||
- | == | + | === Recovery === |
+ | == Sample data == | ||
+ | |||
+ | === Mass spectra === | ||
+ | |||
+ | === Chromatogram === | ||
== Reference == | == Reference == | ||
+ | * PubMed - Ref | ||
+ | |||
+ | [[Category:Flow injection analysis]] | ||
+ | [[Category:Triple quadrupole]] |
Revision as of 15:52, 6 June 2008
How to use:
- Click 'Edit' and copy the source code to clipboard
- Create new page/article and paste the source code into your new article.
- Modify as needed.
Contents |
Sample preparation
- liquid-liquid extraction, SPE ....
- sample sovlent:
Material | Material used | Internal Standard(s) | Internal Standard(s) added |
---|---|---|---|
Cultured cells | 100µg protein | Cer 14:0, Cer 17:0 | 50ng each |
Instrumentation and method
Pump
- Type:
- Mode: isocratic or gradient
- Solvent(s):
- Gradient:
Time [min] | Flow [ml/min] | % Solvent A | % Solvent B |
---|---|---|---|
0 | 0.2 | 100 | 0 |
1.0 | 0.2 | 50 | 50 |
1.5 | 0.2 | 100 | 0 |
Autosampler
- Type:
- Injection volume:
- Wash solvent:
Mass spectrometer
- Type:
- Ionization mode:
- Ionization voltage:
- Source temperature:
- Collision gas: Argon
- Collision gas pressure:
- MS/MS-conditions:
- precursor ion scan of m/z 264.2
- multiple reaction monitoring table of species observed frequently
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
- both [M+H]+ and [M+H-H2O]+ are used
- species used for calibration
Species | Cultured cells | Human plasma |
---|---|---|
Cer 16:0 | 0-35pmol | 0-50pmol |
Cer 18:0 | 0-35pmol | 0-50pmol |
Cer 20:0 | 0-35pmol | 0-50pmol |
Cer 24:1 | 0-35pmol | 0-50pmol |
Cer 24:0 | 0-35pmol | 0-50pmol |
Method validation
Accuracy
Precision
LOD
LOQ
Recovery
Sample data
Mass spectra
Chromatogram
Reference
- PubMed - Ref