Filename ATME98_2_neg.CDF Data format NetCDF Author Dr. Fumio Matsuda Institute LC-MS branch, RIKEN PSC, Japan Date submitting 2009/7/5 Species Arabidopsis thaliana Genotype Col-0 Organ root Biosource amount (mg) 55.16 Growth support 1xMS Salts, 1xMS vitamins, 0.05% MES, 0.8% Agar, pH5.8 with 1M KOH Growth location SANYO Growth chamber Growth plot design approx. 40 seedlings in EIKEN 144x104x16mm polystyrene Light light intensity:60 micromol, luminescence contineous. Humidity 60% Temperature 22C Watering regime no treated Nutritional regime no treated Date(s) of plant establishment 2008/5/24 Other specific metadata nothing special Biotic treatment no treated Abiotic treatment no treated Intervention treatment no treated Treatment dose or intensity levels no treated Treatment time, time intervals and duration before harvest no treated Harvest date, time 2008/6/16 Plant growth stage 21days Metabolism quenching method Freezing with liquid N2 Harvest method Time from tissue resection to liquid N2 freezing was one minute. Tissues were pooled in ependorf tube. Sample strage Stored at -80C for more than one month Reprecate sampling and analyses n=4 Tissue harvesting method Time from tissue resection to liquid N2 freezing was one minute. Tissues were pooled in ependorf tube. Tissue processing method no processing Storage condition prior to extraction or further processing Stored at -80C for more than one month Extraction method The frozen tissues were homogenized in 5 volumes of 80% aqueous methanol containing 0.5 mg/l of lidocaine and d-camphor sulfonic acid (Tokyo Kasei, Tokyo, Japan) by using a mixer mill (MM 300, Retsch) with a zirconia bead for 6 min at 20 Hz. Extract cleanup and/or Additional manipulation Following centrifugation of 15000 g for 10 min and filtration (Ultrafree-MC, 0.2 ?m, Millipore, Bedford, MA, USA) Extract storage and/or relocate Stored at 4 degree for 4 - 48 hours Chromatography instrument description Waters, Acquity UPLC system, operated by MassLynx4.1 Auto-injector Waters, Acquity SM, MassLynx4.1, 2microL injection, 1 wash per injection (600 microL water and 200 microL acetonitrile) Separation column and pre/guard column Waters Acquity BEH C18 (pore size: 1.7 microm) 2.1 by 100 mm with guard column (Waters, VanGuard 2.1 by 5 mm) Separation parameter solvent system: acetonitrile (0.1% formic acid):water (0.1% formic acid); gradient program: 1:99, v/v, at 0 min; 1:99, v/v, at 0.1 min; 99.5:0.5 at 15.5 min; 99.5:0.5 at 17.0 min; 1:99, v/v, at 17.1 min; and 1:99 at 20 min; flow rate: 0.3 ml/min; column temperature: 38 C. Instrument description Waters, Q-Tof Premier, operated by MassLynx4.1 Sample introduction and delivery From LC Ionization source ESI negative. capillary voltage: -3.0 keV; cone voltage: 30 V; source temperature: 120 C; desolvation temperature: 450 C; cone gas flow: 50 l/h; desolvation gas flow: 800 l/h; collision energy: 2 V; Mass analyzer description and acquisition mode quadrupole-time-of-flight operated at scan mode Data acquisition parameters detection mode: scan (m/z 100?2000; dwell time: 0.45 s; interscan delay: 0.05 s, centroid). Lock spray was not employed (mass accuracy: ca. 5ppm, resolution 8500) Data file format NetCDF converted from .RAW data by using DBridge (Waters)