SE199:/MS01

Sample Set Information

ID	SE199
Title	Untargeted metabolome analysis of rice using LC-MS / LC-MSを用いたイネのノンターゲットメタボローム解析
Description	Untargeted metabolome analyses of rice using liquid chromatography-mass spectrometry (LC-MS). Two rice cultivars, Koshichikar and Nipponbare, were used. The information on the peaks detected in rice 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）を用いたイネのノンターゲットメタボローム解析です。試料として、二種類のレタス（コシヒカリとニッポンバレ）を用いました。 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, Keiichi Kanno3, Soichi Kojima3, 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 @) 櫻井望1,2、須田邦裕1、菅野圭一3、小島創一3、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 東北大学農学研究科）
Reference
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