LC-MS/MS parameter Description
HPLC Agilent 1200
Software Version Mass Hunter Version B.01.04
Sample Preparation Plasma Protein precipitation
Sample Matrix Plasma + buffer
Sample Volume 100 µL
Column X Bridge C18 5 µM (4.6 Χ 50 mm)
Mobile Phase (a) 0.1% formic acid in Water
  (b) 0.1% formic acid in Acetonitrile
Flow rate (mL/min) 0.8 mL/min
Column Temperature (°C) 25
Injection Volume (µL) 10
Run time (min) 20
Flow Gradient
QQQ Mass Spectrometer parameter Description
Ion Source Polarity
Delta EMV
Start mass
End mass
Scan mode
Scan time (Dwell)
Fragmentation Voltage
Gas Temperature (°C)
Nebulizer (PSI)
Capillary Voltage
Chamber Current (μA)
Capillary Current (nA)
MS1 Heater (°C)
MS2 Heater (°C)
Turbo1 Speed (%)
Turbo 1 Power (watt)
ESI (Electrospray)
Positive
0
100
1000
EIC (Extracted Ion Chromatogram)
500
135
300
45
4000
0.34
13
100
100
100
104.2
Table 3: Optimized LC-MS/MS parameters.
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