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

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* Fill up the new page/article with your knowledge you have.
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That's it.
 
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== Abstract ==
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== Sample preparation ==
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* Lists are easy to do:
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-
** start every line
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* with a star
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** more stars mean
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*** deeper levels
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== Analysed Matrices ==
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* liquid-liquid extraction, SPE ....
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* sample sovlent:
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{| class="wikitable" style="text-align:center"
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! Material !! Material used  !! Internal Standard(s) !! Internal Standard(s) added
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|-
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! Cultured cells
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| 100µg protein || Cer 14:0, Cer 17:0 || 50ng each
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|}
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== Analytes ==
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== Instrumentation and  method ==
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== Sample Preparation ==
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=== Pump ===
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* Type:
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* Mode: isocratic or gradient
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* Solvent(s):
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* Gradient:
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{| border="1" class="wikitable" style="text-align:center;"
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! Time [min]!! Flow [ml/min]  !! % Solvent A !! % Solvent B
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|-
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! 0
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| 0.2 || 100 ||0
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|-
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! 1.0
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| 0.2 || 50||50
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|-
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! 1.5
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| 0.2 || 100 ||0
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|}
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=== Autosampler ===
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* Type:
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* Injection volume:
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* Wash solvent:
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==Analytical Method==
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=== Mass spectrometer ===
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=== HPLC-MS-MS ===
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* Type:
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* Ionization mode:
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* Ionization voltage:
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* Source temperature:
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* Collision gas: Argon
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* Collision gas pressure:
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* MS/MS-conditions:
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** precursor ion scan of m/z 264.2
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** multiple reaction monitoring table of species observed frequently
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{| border="1" class="wikitable" style="text-align:center"
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! Analyte !! Precursor [m/z] !! Precursor [m/z] !! Collision energy [eV]
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|-
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! species 1
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| m/z 1 || m/z 2||x
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|-
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! species 2
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| m/z 1 || m/z 2||x
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|-
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|}
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==== HPLC (Alliance 2695, Waters) ====
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== Data analysis and quantification ==
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=== Data handling ===
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* chromatogram, spectrum and peak processing
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==== MS (Quattro Ultima MS, Waters) ====
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=== Isotope correction ===  
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* if necessary
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== Method Validation ==
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=== Calibration and quantification ===
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* calibration type: matrix or external calibration by standard addition to sample matrix
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* both [M+H]+ and [M+H-H2O]+ are used
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* species used for calibration
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{| class="wikitable" style="text-align:center"
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! Species!! Cultured cells !! Human plasma
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|-
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! Cer 16:0
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| 0-35pmol || 0-50pmol
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|-
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! Cer 18:0
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| 0-35pmol || 0-50pmol
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|-
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! Cer 20:0
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| 0-35pmol || 0-50pmol
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|-
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! Cer 24:1
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| 0-35pmol || 0-50pmol
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|-
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! Cer 24:0
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| 0-35pmol || 0-50pmol
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|}
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=== LOD ===
 
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== Method validation ==
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=== LOQ ===
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=== Accuracy ===
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=== Precision ===
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=== Linearity ===
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=== LOD ===
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=== LOQ ===
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== Internal Standard (IS) ==
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=== Recovery ===
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== Sample data ==
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=== Mass spectra ===
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=== Chromatogram ===
== Reference ==
== Reference ==
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* PubMed - Ref
 +
 +
[[Category:Flow injection analysis]]
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[[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 5050
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 2x
species 2 m/z 1 m/z 2x

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
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