Category:Method: Difference between revisions
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{{Twocolumn|
{{Twocolumn|
; Homogenize cells or organs with methanol
; Homogenize cells or organs with methanol
** B&D method: [[:Category:Method/Sample/PL/Neutral|Neutral lipids, Phospholipid (PLs)]]
** B&D method: [[:Category:Method/Sample/PL/Neutral|Neutral lipids, Phospholipids (PLs)]]
** Solid-phase extraction
** Solid-phase extraction
*** Reverse column C18 : [[:Category:Method/Sample/PL/Oxidized|Oxidized PLs, Oxidized fatty acids]]
*** Reverse column C18 : [[:Category:Method/Sample/PL/Oxidized|Oxidized PLs, Oxidized fatty acids]]
*** Ion-exchange column DEAE : [[:Category:Method/Sample/PL/Acidic_PI|Phosphoinositides]]
*** Ion-exchange column DEAE : [[:Category:Method/Sample/PL/Acidic_PI|Phosphoinositides]]
** Chloroform-methanol extraction (Ledeen method) : [[:Category:Method/Sample/GL/Acidic|Glycolipid(Gangliosides, Sulphatides)]]
** Chloroform-methanol extraction (Ledeen method) : [[:Category:Method/Sample/GL/Acidic|Glycolipids(Gangliosides, Sulphatides)]]
; Butanol extraction  
; Butanol extraction  
** Solid-phase, Ion-exchange column DEAE : [[:Category:Method/Sample/PL/Acidic|Acidic PLs]]
** Solid-phase, Ion-exchange column DEAE : [[:Category:Method/Sample/PL/Acidic|Acidic PLs]]

Revision as of 04:37, 22 October 2010

Top Lipid Class Method Symbols

Overview

Lipid contents exhibit high dynamic range. Detection of scarce elements is difficult as it is due to ion suppressions; different extraction methods are necessary depending on purposes.

  1. Sample preparation ... extraction depending on purposes
  2. Analytical strategy ... selection of platforms and methods
  3. Computational analysis ... qualitative/quantitative and multivariate analysis

脂質は量的に多い成分と少ない成分のダイナミックレンジが大きいため、主要成分と混合したままだと微量成分の検出はイオンサプレッションなどにより難しくなる。つまり、目的に応じたサンプル調製が重要である。

  1. 前処理 ... 目的に応じたサンプル調製
  2. 分析 ... 装置と測定方法の選択
  3. 数理解析　... 定性と定量ならびに多変量解析

Amount & Species
abundant      few
LipidContents.png
scarce      many

Sample Preparation

Homogenize cells or organs with methanol
Butanol extraction
    • Solid-phase, Ion-exchange column DEAE : Acidic PLs

細胞・臓器をメタノールでホモジナイズ
ブタノール抽出法

Analytical Strategy

Strategy Method Type Instrument
Global (detection of all lipids) LC/MS
Data dependent scan (LC/MS/MS, LC/MS3)

Hybrid type MS

quadrupole-time-of-flight (Q-TOF)
  • Q-TOF (Waters)

Hybrid type MS

ion-trap (IT)-TOF or Fourier-Transform Ion Cyclotron Resonance (FT-ICR)
  • LTQ-Orbitrap (Theremo-Fisher Scientific)
Focused (focusing on partial structures) Precursor ion scan
Neutral loss scan 		Triple-quadrupole MS
  • 4000 Q-TRAP or 5500 Q-TRAP (Applied Biosystems)
  • TSQ Quantum or Vantage (Thermo- Fisher Scientific)
Targeted (targeting by molecular and fragment ions) Multiple reaction monitoring (MRM),
Selected reaction monitoring(SRM)

Computational Analysis

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