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Arabidopsis Metabolome Analysis Using Infusion ESI FT-ICR/MS

融合ESI FT-ICR/MS法分析拟南芥代谢组

Reiko Motohashi Reiko Motohashi
MS Masakazu Satou
Fumiyoshi Myouga Fumiyoshi Myouga
Akira Oikawa Akira Oikawa
DO Daisaku Ohta

发布: 2015年05月05日第5卷第9期 DOI: 10.21769/BioProtoc.1463 浏览次数: 8917

评审: Arsalan DaudiSekhar KambakamAnonymous reviewer(s)

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Abstract

We made the method for Arabidopsis metabolome analysis based on direct-infusion Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR/MS) (IonSpec). This method was sufficiently applied to metabolic phenotyping of Arabidopsis. This method is simple in that after homogenizing samples, powdered samples are dissolved in extraction solvents (acetone and methanol) to 20% fresh weight/volume. Extracted sample solutions are dried and dissolved in 50% (v/v) acetonitrile. Mass analysis using FT-ICR/MS (IonSpec) is performed in positive and negative ionization operation modes. Mass spectra are acquired over the 100-1,000 m/z range and accumulated to improve the S/N ratio.

Keywords: Metabolome (代谢组学) search Arabidopsis (拟南芥) search Albino (白化) search ESI FT-ICR/MS (ESI FT-ICR质谱) search

Materials and Reagents

  1. 3-week-old Arabidopsis plants
  2. Liquid nitrogen
  3. Acetone (HPLC grade) (Wako Chemicals USA, catalog number: 014-08681 )
  4. Methanol (HPLC grade) (Wako Chemicals USA, catalog number: 134-14523 )
  5. Nitrogen gas
  6. EN1-16 (TAITEC, catalog number: 0076417-000 )
  7. Formic acid (HPLC grade) (Wako Chemicals USA, catalog number: 063-04192 )
  8. Lidocaine (anaesthetic) m/z 235.18104 (Wako Chemicals USA, catalog number: 120-02691 )
  9. Prochloraz (agricultural chemical) m/z 376.03863 (Wako Chemicals USA, catalog number: 164-25131 )
  10. Reserpine (alkaloid sedative drug) m/z 609.28121 (Wako Chemicals USA, catalog number: 184-00691 )
  11. Bombesin (peptide) m/z 810.41479 (Wako Chemicals USA, catalog number: 339-40861 )
  12. 28~30% ammonia solution (Wako Chemicals USA, catalog number: 016-03146 )
  13. Negative mode internal standards: 2.4-D (plant hormone) m/z 218.96157 (Wako Chemicals USA, catalog number: 040-18532 )
  14. Ampicillin (antibiotic) m/z 348.10180 (Wako Chemicals USA, catalog number: 017-20531 , CHAPS 8 detergent) m/z 613.388865 (Wako Chemicals USA, catalog number: 341-04721 )
  15. Tetra-N-acetylchitotetraose [(GluNAc)4] m/z 829.32023 (Tokyo Chemical Industry, catalog number: T2910 )
  16. 50% (v/v) acetonitrile (HPLC grade) (Wako Chemicals USA, catalog number: 018-19853 ) [use distilled water to dilute acetonitrile to 50%(v/v)]
  17. Acetic acid (Wako Chemicals USA, catalog number: 014-20063 )
  18. 28~30% ammonia solution (Wako Chemicals USA, catalog number: 016-03146 )
  19. Ultrapure water (recommended but not required)

Equipment

  1. An IonSpec Explorer FT-ICR/MS equipped with a 7-tesla actively shielded superconducting magnet (Agilent, Ionspec)
  2. Glass vial and teflon cap (GL Biochem, catalog number: 1030-46716 )
  3. Glass pipette
  4. 0.45 µm filters (PTFE) (ADVANTEC®, catalog number: 13HP045AN )
  5. Heat block

Procedure

English
中文翻译

文章信息

版权信息

© 2015 The Authors; exclusive licensee Bio-protocol LLC.

如何引用

Motohashi, R., Satou, M., Myouga, F., Oikawa, A. and Ohta, D. (2015). Arabidopsis Metabolome Analysis Using Infusion ESI FT-ICR/MS. Bio-protocol 5(9): e1463. DOI: 10.21769/BioProtoc.1463.

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分类

植物科学 > 植物生物化学 > 其它化合物

植物科学 > 植物新陈代谢 > 代谢物谱

系统生物学 > 代谢组学 > 全生物体

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