Sample Set Information
|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 @)
|Title||BY2 cells, 3 days|
|Organism - Scientific Name||Nicotiana tabacum|
|Organism - ID||NCBI taxonomy: 4097|
|Compound - ID|
|Compound - Source|
|Sample Preparation Details ID||SS1|
Sample Preparation Details Information
|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.|
Analytical Method Information
|Title||ESI Negative (Method 5)|
|Method Details ID||MS11|
|Sample Amount||5 mg FW / 20 uL injection|
Analytical Method Details Information
|Title||LC-FT-MS, ESI, Negative|
|Instrument||Agilent1100 HPLC (Agilent), LTQ-FT (Thermo Fisher Scientific)|
|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.
|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