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Harvested sample is frozen by liquid N2 an … 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. 32.98011, 235.18048, 249.20600, 266.23260.
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