SE44:/S04/M02

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Sample Set Information

ID SE44
Title Physcomitrella patens metabolite analysis using stable isotopes
Description Metabolites in the Physcomitrella patens are investigated by using stable isotope labelings. This analysis was performed to estimate the elemental composition with high reliability using the number of stable isotopes incorporated in the compound as an index and to add information to discuss the biosynthetic pathway. The Physcomitrella patens were labeled with 13C, 15N, 18O, and 34S, respectively in the same condition. Following peak picking by using PowerGet, the metabolite list was prepared from unlabeled data and blank data. The isotopic peaks were searched and listed using ShiftedIonsFinder. Moreover, the flavonoid-like peaks were also searched using ShiftedIonsFinder.
Authors Yasuhide Hiraga 1,2, Takeshi Ara 1, Yoshiki Nagashima 1, Nozomu Sakurai 1, Hideyuki Suzuki 1,2, Kota Kera 1,3; 1:Kazusa DNA Research Institute, Japan, 2:Hirata Corporation, Japan, 3:Faculty of Applied Bioscience, Tokyo University of Agriculture, Japan
Reference Hiraga Y, et al, (2020) Plant Biotechnology, 37(3), 377-381.
Comment


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Sample Information

ID S04
Title Physcomitrella patens subsp. patens
Organism - Scientific Name Physcomitrella patens subsp. patens
Organism - ID NCBI taxonomy:3218
Compound - ID
Compound - Source
Preparation
Sample Preparation Details ID SS1
Comment

Sample Preparation Details Information

ID SS1
Title Sealed-glass bottle system
Description The gametophore of P. patens were grown on solid (0.5 % phytagel) half strength Murashige-Skoog (MS) medium containing 0.5 % glucose at 22°C under 18 h-light/6 h-dark long day condition using sealed-glass bottle system for 2 months. Gametophores were harvested as three biological replicates, frozen in liquid nitrogen, and then lyophilized.
Comment_of_details

Analytical Method Information

ID M02
Title LC-Orbitrap-MS, ESI Positive analysis
Method Details ID MS3
Sample Amount 1.7 mg
Comment [MassBase ID] MDLC1_40785


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The raw (binary) and near-raw (text) files of this analysis are available at MassBase.

Analytical Method Details Information

ID MS3
Title LC-Orbitrap-MS ESI positive method 3
Instrument Agilent1200 HPLC (Agilent), LTQ-Orbitrap (Thermo Fisher Scientific)
Instrument Type LC-Orbitrap-MS
Ionization ESI
Ion Mode Positive
Description Harvested sample is frozen by liquid N2 and resulting powder (100mg) are solved in 300uL 80% methanol solution. 5uL sample is injected into HPLC after 0.2um membrane filter treatment. HPLC conditions: Agilent 1200 series (Agilent), Column: TSKgel ODS-100V (4.6 x 250 mm, 5 micrometer; TOSOH), Solvent: A; 0.1% formic acid aq. B; ACN (addition 0.1% formic acid fc.), Gradient: (B) 3 to 97% for 45 min, 97% for 5 min and 97 to 3% for 7 min, Column temp.: 40 degree C, Flow rate=0.5mL/min, PDA: 190-950 nm (2 nm step). Orbitrap-MS conditions: Filter 1: FTMS + c norm !corona !pi res=60000 o(100.0-1500.0); 2: ITMS + c norm !corona !pi Dep MS/MS Most intense ion from (1); 3: ITMS + c norm !corona !pi Dep MS/MS 2nd most intense ion from (1); 4: ITMS + c norm !corona !pi Dep MS/MS 3rd most intense ion from (1); 5: ITMS + c norm !corona !pi Dep MS/MS 4th most intense ion from (1); 6: ITMS + c norm !corona !pi Dep MS/MS 5th most intense ion from (1). Reject Mass List:102.09139, 110.02006, 141.95860, 158.00286, 158.96135, 163.13289, 171.14921, 182.98517, 186.95628, 195.08767, 199.98790, 207.15909, 223.06365, 224.18561, 227.98281, 232.98011, 235.18048, 249.20600, 266.23260.
Comment_of_details 20130424_Standerd_ESp.meth
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