System Parameter Setting
Autosampler Temperature 4°C
  Injection 5.0µL prepared sample
LC Trap Column Sample load time 5 min
  Trap flow rate 5 µL/min
LC Analytical Column Column temperature 55 °C
  Flow rate 0.4µL/min
  Gradient Initial conditions: 98% A
    t = 60 min, 60% A
    t = 65 min, 5% A
    t = 70 min, 5 % A
    t = 75 min, 98 % A
    t = 95 min, 98 % A
LTQ Orbitrap XL Mass Spectrometer Experiment type Nth Order Double Play
  # Dependent scans 5
  Source voltage 2.0 kV
  Capillary temperature 200 °C
  Source current 1.0 µA
  Capillary voltage 47.0 V
  Tube lens 100.0 V
  FTMS survey scan time 1.0 sec
  FTMS resolution 60,000
  Survey time 90 min
  Survey ion mode ES Mode
  Survey polarity Positive
  Survey mass range 375-2000
  Activation type CID
  Normalized collision energy 35.0
  ITMS MS/MS mass range 50-2000
  ITMS MS/MS scan time 0.2 sec
Table 1: LC and MS Instrument Parameters used in the study: Analytical samples were analyzed on a Thermo Scientific LTQ Orbitrap XL mass spectrometry system coupled to a Waters nano ACQUITY UPLC system using parameters presented in the Table below.