SE204:/S07/M014
Sample Set Information
ID | SE204 |
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Title | Untargeted metabolome analysis of tobacco (BY-2) cells using LC-MS / LC-MSを用いたタバコ(BY-2)培養細胞のメタボローム解析 |
Description | Untargeted metabolome analyses of tobacco (BY-2) cell cultures using liquid chromatography-mass spectrometry (LC-MS).
The information on the peaks detected in this study was available at the Plant Metabolome Repository website (http://metabolites.in/plants).
検出されたピークは、植物メタボロームレポジトリ (http://metabolites.in/plants) から公開されています。 |
Authors | Sakurai N1,2, Suda K1, Masumoto H1, Kugou K1, Akimoto N1, Hoshi K1, Osawa S1, Ikeda C1, Ozawa K1, Yamada M1, Muneto R1, Shibata D1 (1 Kazusa DNA Research Institute, 2 National Institute of Genetics), Contact: sakurai AT nig.ac.jp (replace AT with @)
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Reference | |
Comment |
Sample Information
ID | S07 |
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Title | BY2 cells, 7 days |
Organism - Scientific Name | Nicotiana tabacum |
Organism - ID | NCBI taxonomy: 4097 |
Compound - ID | |
Compound - Source | |
Preparation | |
Sample Preparation Details ID | SS1 |
Comment |
Sample Preparation Details Information
ID | SS1 |
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Title | Preparation of the samples |
Description | The tobacco BY-2 cells were provided by the RIKEN BRC through the National Bio-Resource Project of MEXT, Japan. The cells were cultured according to a general protocol. The cells or culture media were sampled after 0, 3, or 7 days after inoculation. The samples were immediately frozen in liquid nitrogen, and stored at -80C. The cell sample was homogenated using mortar and pestle under liquid nitrogen into a fine powder and stored at -80C until use. |
Comment_of_details |
Analytical Method Information
ID | M014 |
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Title | ESI Positive (Method 1) |
Method Details ID | MS01 |
Sample Amount | 5 mg FW / 20 uL injection |
Comment |
Analytical Method Details Information
ID | MS01 |
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Title | LC-FT-MS, ESI, Positive |
Instrument | Agilent1100 HPLC (Agilent), LTQ-FT (Thermo Fisher Scientific) |
Instrument Type | LC-FTICR-MS |
Ionization | ESI |
Ion Mode | Positive |
Description | Metabolite extraction
300 mg of powdered sample was mixed with 900 uL of 100% methanol solution containing 25 uM of 7-hydroxy-5-methylflavone as internal standard (IS). The sample was homogenized using Mixer Mill MM 300 (QIAGEN) at 25 Hz for 2 min, twice. The homogenate was centrifuged under 17,400 x g for 5 min at 4 C. A supernatant was passed through a Polytetrafluoroethylene (PTFE) filter (pore size 0.2 um, Millipore), and the filtrate was applied to a C18 silica column (MonoSpin C18, GL Science) to remove highly-hydrophobic contaminants. The filtrate passed through the C18 column was used for LC-MS analyses. The same extraction procedure with 75% methanol was performed without a sample to prepare mock samples and used as negative controls.
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Comment_of_details | [column] TSK-gel ODS-100V (4.6 x 250 mm, 5 micrometer; TOSOH)
[gradient] Solvent A: water containing 0.1% v/v formic acid; Solvent B: acetonitrile containing 0.1% v/v formic acid; Gradient: 3% B (0 min), 97% B (90 min), 97% B (100 min), 3% B (100.1 min), and 3% B (107 min) [total separation time] 107 min |