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Table 2 Optimized parameters of the established LC–MS/MS methodology

From: Identification and characterization of in vivo, in vitro and reactive metabolites of vandetanib using LC–ESI–MS/MS

LC parameters MS/MS parameters
HPLC Agilent 1200 (Agilent Technologies, CA, USA) Mass spectrometer Agilent 6410 QqQ (Agilent Technologies, CA, USA)
Mobile phase (gradient) Aqueous phase: 10 mM Ammonium formate in H2O (pH: 4.1 using Formic acid) Ionization source Positive electrospray ionization (ESI)
Organic phase: ACN (0.1% Formic acid) Drying gas: N2 gas
Pressure (55 psi)
Flow rate (12 L/min)
Flow rate: 0.2 mL/min
Elution time: 90 min
Agilent eclipse plus C18 Column   In vitro In vivo ESI temperature: 350 °C
Length (mm) 50 150 Capillary voltage: 4000 V
Internal diameter (mm) 2.1 2.1 Collision gas N2 (high purity)
Particle size (μm) 1.8 3.5 Modes Product ion (PI) and full mass scan and
Temperature: 22 ± 2 °C Software Mass Hunter software
Elution system Time (min) %B (ACN) Analyte Vandetanib, in vivo, in vitro and reactive metabolites
0 5
60 25 Mass conditions Fragmentor voltage (V) 140
80 70
90 5
Post time 15 5 Collision energy (eV) 15