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=={{Bilingual|生体試料の調整|Sample Preparation}}==
{{Taguchi/Header}}
 
=={{Bilingual|概要|Overview}}==
{|
|
{{Twocolumn|
{{Twocolumn|
Sample Preparation details for phospholipids (PL) and glycolipids (GL).
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
|
|
リン脂質 (PL) および糖脂質 (GL) のサンプル調整法
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
}}
}}
{{#def:separator|Sample/}}
|align="center"|
{{#repeat:LinkCategoryPage|1|
<small>'''{{Bilingual|含有量と分子種|Amount & Species}}'''<br/>
{{#SearchTitle:Method/Sample|Category}}
{{Bilingual|多い &nbsp;&nbsp;&nbsp;&nbsp;少ない|abundant &nbsp;&nbsp;&nbsp;&nbsp; few}}<br/>
[[Image:LipidContents.png|100px]]<br/>
{{Bilingual|少ない &nbsp;&nbsp;&nbsp;&nbsp;多い|scarce &nbsp;&nbsp;&nbsp;&nbsp; many}}
</small>
|}
=={{Bilingual|前処理|Sample Preparation}}==
{{Twocolumn|
; Strategy 1. Homogenize cells or organs with methanol
# B&D method: [[:Category:Method/Sample/PL/Neutral|Neutral lipids, Phospholipids (PLs)]]
# Solid-phase extraction
## Reverse column C18 : [[:Category:Method/Sample/PL/Oxidized|Oxidized PLs, Oxidized fatty acids]]
## Ion-exchange column DEAE : [[:Category:Method/Sample/PL/Acidic_PI|Phosphoinositides]]
# Chloroform-methanol extraction (Ledeen method) : [[:Category:Method/Sample/GL/Acidic|Glycolipids(Gangliosides, Sulphatides)]]
; Strategy 2. Butanol extraction
# Solid-phase extraction using ion-exchange column DEAE-cellulose column [[:Category:Method/Sample/PL/Acidic|Acidic PLs (PA, LPA, PS, LPS)]]
|
;手法その1 細胞・臓器をメタノールでホモジナイズ
# B&D法 : [[:Category:Method/Sample/PL/Neutral|中性脂質、リン脂質、]]
# 固相抽出法
## 逆相カラム(C18) : [[:Category:Method/Sample/PL/Oxidized|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) : [[:Category:Method/Sample/PL/Acidic_PI|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) : [[:Category:Method/Sample/GL/Acidic|酸性糖脂質(ガングリオシド、スルファチド)]]
;手法その2 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 : [[:Category:Method/Sample/PL/Acidic|酸性リン脂質(PA, LPA, PS, LPS)
]]
}}
}}


=={{Bilingual|分析法|Analytical Category}}==
=={{Bilingual|分析|Analytical Strategy}}==
{{#def:separator|Analytical/}}
 
{{#repeat:LinkCategoryPage|1|
{| class="wikitable"
{{#SearchTitle:Method/Analytical|Category}}
! Strategy || Method || Type || Instrument
}}
|-
|rowspan="2"| '''Global''' ({{Bilingual|ターゲットを絞らない網羅的手法|detection of all lipids}})
|rowspan="2"| LC/MS <br/> Data dependent scan (LC/MS/MS, LC/MS3)
|
''Hybrid type MS''
: {{Bilingual|四重極(Q)と飛行時間(TOF)のハイブリッド|quadrupole-time-of-flight (Q-TOF)}}
|
* Q-TOF (Waters)
|-
|
''Hybrid type MS''
: {{Bilingual|イオントラップ(IT)とTOFまたはフーリエ変換(FT)のハイブリッド| ion-trap (IT)-TOF or Fourier-Transform Ion Cyclotron Resonance (FT-ICR)}}
|  
* LTQ-Orbitrap (Theremo-Fisher Scientific)
|-
| '''Focused''' ({{Bilingual|共通構造部分に焦点をあてた手法|focusing on partial structures}})
| Precursor ion scan<br/> Neutral loss scan
|rowspan="2"| {{Bilingual|三連四重極(TSQ)MS
| Triple-quadrupole MS}}
|rowspan="2"|
* 4000 Q-TRAP or 5500 Q-TRAP (Applied Biosystems)
* TSQ Quantum or Vantage (Thermo- Fisher Scientific)
|-
| '''Targeted''' ({{Bilingual|個別分子特異的な手法|targeting by molecular and fragment ions}})
| Multiple reaction monitoring (MRM),<br/> Selected reaction monitoring(SRM)
|}
 
=={{Bilingual|数理解析|Computational Analysis}}==
 
* [http://lipidsearch.jp/ LipidSearch]
* [http://masspp.jp/ Mass++]
* [http://www.infocom.co.jp/bio/develop/simca-p_plus.html SIMCA-P]
* [http://www.absciex.com/ Marker View]
* [http://www.thermoscientific.jp/lc-lcms/sieve.html SEIVE]
* [http://www.ncc.go.jp/jp/nccri/divisions/03chem/pdf/2DICAL.pdf 2DICAL]

Latest revision as of 11:12, 5 November 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

Strategy 1. Homogenize cells or organs with methanol
  1. B&D method: Neutral lipids, Phospholipids (PLs)
  2. Solid-phase extraction
    1. Reverse column C18 : Oxidized PLs, Oxidized fatty acids
    2. Ion-exchange column DEAE : Phosphoinositides
  3. Chloroform-methanol extraction (Ledeen method) : Glycolipids(Gangliosides, Sulphatides)
Strategy 2. Butanol extraction
  1. Solid-phase extraction using ion-exchange column DEAE-cellulose column Acidic PLs (PA, LPA, PS, LPS)

手法その1 細胞・臓器をメタノールでホモジナイズ
  1. B&D法 : 中性脂質、リン脂質、
  2. 固相抽出法
    1. 逆相カラム(C18) : 酸化リン脂質、酸化脂肪酸
    2. イオン交換カラム(DEAEセルロース) : ホスホイノシタイド
  3. 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)
手法その2 ブタノール抽出法
  1. 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)

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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