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Chromatographical separation of metabolites was performed on Syncronis HILIC UHPLC column (2.1 mm 100 mm 1.7 m, Thermo Fisher) at the flow rate of 600 L/min using a binary solvent system: solvent A, 20 mM ammonium formate, pH 3.0 in mass spectrometry grade H 2 0, and solvent B, mass spectrometry grade acetonitrile with 0.1% formic acid (%v/v)

Each escalation phase requires recalculating injection volume unless you prepare a reference chart

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Having protein-forward meals and snacks will help keep you fuller longer and make it easier to snack less throughout the day