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The MS1 spectrum was acquired (Orbitrap analysis, resolution 60,000, 350-1350 Th, automatic gain control (AGC) target 100%, maximum injection time 118 ms), for ~90 min per fraction, followed by the high-resolution MS2 stage consisting of fragmentation by higher energy collisional dissociation (HCD, normalized collision energy 35%) and analysis using the Orbitrap (AGC 200%, maximum injection time 120 ms, isolation window 0.6 Th, resolution 50,000)

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Encouragingly, hybrid systems often exhibit reduced fed/fasted variability, a key requirement for regulatory biowaivers under BCS Class II guidelines [134]