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Prepared the mobile phase solution, A (0.1% (v/v) formic acid in ddH 2 O), B (0.1% (v/v) formic acid and 80% acetonitrile in ddH 2 O), the lyophilized samples were dissolved with 10 L solution A, then centrifuged at 14000 g for 20 min at 4 C, 400 ng supernatant of the samples were injected onto a Thermo Scientific UltiMate 3000 UHPLC coupled to a timsTOF HT mass spectrometer (BRUKER)

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The length of this phase depends on how much weight you have to lose