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Sample is frozen by liquid N2 and resultin … Sample is frozen by liquid N2 and resulting powder (100mg) are solved in 300uL 80% methanol solution. 20uL sample is injected into HPLC after 0.2um membrane filter treatment. HPLC conditions: Agilent 1100 series (Agilent), Column: TSKgel-100Z (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 30% (0.0 to 25.0 min), 30 to 90% (25.1 to 40.0 min), 95% (45.1 to 50.0 min), 3% (50.1 to 57.0 min), Column temp.: 30 degree C, Flow rate=0.5mL/min, PDA: 200-650 nm (2 nm step). FT-ICR-MS conditions: Filter 1: FTMS + c norm !corona !pi res=100000 o(50.0-2000.0); 2: ITMS + c norm !corona !pi Dep MS/MS Most intense ion from (1); 3: ITMS + c norm Dep MS/MS 2nd most intense ion from (1); 4: ITMS + c norm Dep MS/MS 3rd most intense ion from (1); 5: ITMS + c norm Dep MS/MS 4th most intense ion from (1); 6: ITMS + c norm Dep MS/MS 5th most intense ion from (1), Reject Mass List:224.1900, 249.2100, 266.2300, 294.2600, 554.1800, 756.5500, 782.5700, 783.5700, 804.5500 , 950.8100. , 782.5700, 783.5700, 804.5500 , 950.8100.
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