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

From LipidomicsWiki

(Difference between revisions)
Jump to: navigation, search
Current revision (16:00, 3 December 2008) (view source)
m
 
(28 intermediate revisions not shown.)
Line 1: Line 1:
-
How to use:
+
== Sample preparation  ==
-
* 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.
+
 +
Material:  Palmoplantar stratum corneum scrapings from healthy volunteers
-
== Sample preparation ==
+
Extraction:  Chloroform–Methanol (1:1, v/v)
-
* liquid-liquid extraction, SPE ....
+
== Instrumentation and method  ==
-
* sample sovlent:
+
-
{| class="wikitable" style="text-align:center"
+
=== Pump ===
-
! Material !! Material used !! Internal Standard(s) !! Internal Standard(s) added
+
-
|-
+
-
! Cultured cells
+
-
| 100µg protein || IS1, IS2 || xng each
+
-
|-
+
-
! Human plasma
+
-
| 10µl || IS1, IS2  || xng each
+
-
|}
+
-
== Instrumentation and  method ==
+
*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"
-
=== Pump ===
+
|-
-
* Type:
+
! Time [min]  
-
* Mode: isocratic or gradient
+
! Flow [ml/min]  
-
* Solvent(s):
+
!  % Solvent A  
-
* Gradient:
+
!  % Solvent B
-
{| border="1" class="wikitable" style="text-align:center;"
+
-
! Time [min]!! Flow [ml/min] !! % Solvent A !! % Solvent B
+
|-
|-
! 0  
! 0  
-
| 0.2 || 100 ||0
+
| 1
 +
| 100  
 +
| 0
|-
|-
-
! 1.0
+
! 15
-
| 0.2 || 50||50
+
| 1  
-
|-
+
| 0  
-
! 1.5
+
| 100
-
| 0.2 || 100 ||0
+
|}
|}
-
=== Column ===
+
=== Column ===
-
if used
+
-
* Type:
+
-
* Temperature:
+
-
 
+
-
=== Autosampler ===
+
-
* Type:
+
-
* Injection volume:
+
-
* Wash solvent:
+
-
 
+
-
=== Mass spectrometer ===
+
-
* Type:
+
-
* Ionization mode:
+
-
* Ionization voltage:
+
-
* Source temperature(s):
+
-
* Collision gas:
+
-
* Collision gas pressure:
+
-
* MS/MS-conditions:
+
-
** multiple reaction monitoring table
+
-
{| 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
+
-
* species used for calibration
+
-
{| class="wikitable" style="text-align:center"
+
-
! 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 ==
+
*Type: silica column LiChroCart 125-4 with Si 60 (5µm) (merck)
-
=== Mass spectra ===
+
=== Mass spectrometer  ===
-
=== Chromatogram ===
+
*Type: Finnigan LCQ classic (ion trap mass spectrometer)
 +
*Ionization mode: APCI, positive full scan mode
 +
*Source temperature(s):&nbsp; APCI heater:&nbsp; set to 500°C,&nbsp; heated capillary:&nbsp;150°C<br>
-
== Reference ==
+
<br>
-
* PubMed - Ref or electronic journal
+
 +
== Reference  ==
-
Please add categories as below for:
+
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&nbsp;(2003) 233-239.
-
* analytical techniques applied
+
-
* lipid classes analyzed (see lipid class categories)
+
-
[[Category:Lipid mass spectrometry]]
+
[[Category:APCI]] [[Category:Single_quadrupole]] [[Category:Ceramides_(SP02)]] [[Category:LC-MS]]

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.

Personal tools
Create a book