SE205:/S06/M05

Sample Set Information

ID	SE205
Title	Untargeted metabolome analysis of plants (6 species) using LC-MS / LC-MSを用いた植物（6種）のノンターゲットメタボローム解析
Description	Untargeted metabolome analyses of plants (6 species) 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). 液体クロマトグラフィー-質量分析（LC）を用いた植物（6種類）のノンターゲットメタボローム解析です。 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, Hasebe M3, Ishikawa M3, Kohchi T4, Nishihama R4, Nakai A4, Demura T5, Sano R5, Ohtani M6, Ihara A6, Susa F7, 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, 3 National Institute of Basic Biology, 4 Kyoto Univ., 5 Nara Institute of Science and Technology, 6 Tokyo Univ., 7 RIKEN CSRS), Contact: sakurai AT nig.ac.jp (replace AT with @) 櫻井望1,2、須田邦裕1、長谷部光泰3、石川雅樹3、河内孝之4、西浜竜一4、中井綾4、出村拓5、佐野亮介5、大谷美沙都6、井原あゆみ6、須佐文子7、秋元奈弓1、星久美1、大澤祥子1、池田千晶1、小澤馨史1、山田学1、宗藤玲子1、柴田大輔1（1 かずさDNA研究所, 2 国立遺伝学研究所、3 基礎生物学研究所、4 京都大学、5 奈良先端大学、6 東京大学、7 理研CSRS）
Reference
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Sample Information

ID	S06
Title	Marchantia polymorpha, culture medium
Organism - Scientific Name	Marchantia polymorpha
Organism - ID	NCBI taxonomy: 3197
Compound - ID
Compound - Source
Preparation
Sample Preparation Details ID
Comment

Analytical Method Information

ID	M05
Title	ESI Positive (Method 5)
Method Details ID	MS01
Sample Amount	5 mg FW / 20 uL injection
Comment

Analytical Method Details Information

ID	MS01
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. Liquid chromatography (LC)-mass spectrometry (MS) analysis Agilent 1100 system (Agilent) and Finnigan LTQ-FT (Thermo Fisher Scientific) were used. Aliquots (5 uL) of the methanol extract were applied to a TSK-gel ODS-100V column (4.6 x 250 mm, 5 um, TOSO) and separated by water containing 0.1%(v/v) formic acid (Solvent A) and acetonitrile containing 0.1%(v/v) formic acid (Solvent B). The gradient program was as follows: 3% B (0 min), 97% B (90 min), 97% B (100 min), 3% B (100.1 min), and 3% B (107 min). The flow rate was set at 0.25 mL/min for 0-100 min and 0.5 mL/min for 100.1-107 min. The conditions of mass spectrometry were as below: - Method 1: This method is used for measuring the accurate mass of the peaks and obtaining MS/MS spectra. [Full scan] resolution, 100,000 by FT; m/z range, 100-1500 [MS2 scan] data-dependent scan by IT for the most intense 5 ions in the full scan. [Dynamic exclusion setting for MS2 scan] repeat count, 3; repeat duration, 30 s; exclusion list size, 500; margin, 10 ppm; exclusion duration, 20 s. - Method 2: Same as Method 1 except below: [Dynamic exclusion setting for MS2 scan] exclusion duration, 30 s. - Method 3: Same as Method 1 except below: [Dynamic exclusion setting for MS2 scan] repeat count, 2; exclusion duration, 30 s. - Method 4: Same as Method 1 except below: [Dynamic exclusion setting for MS2 scan] off. - Method 5: This method is used for obtaining MS3 spectra. [Full scan] resolution, 12,500 by FT; m/z range, 100-1500. [MS2 scan] data-dependent scan by IT for the most intense 5 ions in the full scan. [Dynamic exclusion setting for MS2 scan] same as Method 1. [MS3 scan] data-dependent scan by IT for the most intense 2 ions in the MS2 scan - Method 6: This method is used for obtaining MS2 spectra in high-resolution: [Full scan] resolution, 12,500 by FT; m/z range, 100-1500. [MS2 scan] data-dependent scan by FT for the most intense 5 ions in the full scan. resolution, 12,500. [Dynamic exclusion setting for MS2 scan] repeat count, 1; repeat duration, 30 s; exclusion list size, 500; margin, 10 ppm; exclusion duration, 40 s. - Method 7: Same as Method 5 except below: [Dynamic exclusion setting for MS2 scan] same as Method 2 ESI: electrospray ionization, FT: Fourier transformation ion cyclotron resonance type mass spectrometry, IT: ion trap type mass spectrometry. The mass analyses with electrospray ionization (ESI) in positive or negative modes were performed. The combinations of Methods 1-7 and ionization polarity are different between the samples, but at least one method for accurate mass measurement (Method 1-4) was applied for each sample. The raw data were acquired using Xcalibur software (Thermo Fisher Scientific).
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