Category:Method: Difference between revisions

Revision as of 08:50, 26 October 2010

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.

Sample preparation ... extraction depending on purposes
Analytical strategy ... selection of platforms and methods
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。

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

Amount & Species
abundant few

scarce many

Sample Preparation

Homogenize cells or organs with methanol

- B&D method: Neutral lipids, Phospholipids (PLs)
- Solid-phase extraction
  - Reverse column C18 : Oxidized PLs, Oxidized fatty acids
  - Ion-exchange column DEAE : Phosphoinositides
- Chloroform-methanol extraction (Ledeen method) : Glycolipids(Gangliosides, Sulphatides)

Butanol extraction

- Solid-phase extraction using ion-exchange column DEAE-cellulose column Acidic PLs (PA, LPA, PS, LPS)

細胞・臓器をメタノールでホモジナイズ

- B&D法 : 中性脂質、リン脂質、
- 固相抽出法
  - 逆相カラム（C18） : 酸化リン脂質、酸化脂肪酸
  - イオン交換カラム（DEAEセルロース） : ホスホイノシタイド
- クロロホルムーメタノール抽出法（Ledeen法の改変） : 酸性糖脂質（ガングリオシド、スルファチド）

ブタノール抽出法

- イオン交換カラム（DEAEセルロース）を用いた固相抽出法 : 酸性リン脂質（PA, LPA, PS, LPS）

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)
Global (detection of all lipids)	LC/MS Data dependent scan (LC/MS/MS, LC/MS3)	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)	Triple-quadrupole MS

Computational Analysis

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@@ Line 31: / Line 31: @@
 ** Chloroform-methanol extraction (Ledeen method) : [[:Category:Method/Sample/GL/Acidic|Glycolipids(Gangliosides, Sulphatides)]]
 ; Butanol extraction
-** Solid-phase extraction using ion-exchange column DEAE-cellulose column [[:Category:Method/Sample/PL/Acidic|Acidic PLs]]
+** Solid-phase extraction using ion-exchange column DEAE-cellulose column [[:Category:Method/Sample/PL/Acidic|Acidic PLs (PA, LPA, PS, LPS)]]
 |
 ; 細胞・臓器をメタノールでホモジナイズ

Category:Method: Difference between revisions

Revision as of 08:50, 26 October 2010

Contents

Overview

Sample Preparation

Analytical Strategy

Computational Analysis

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