Table 2 Adjusted parameters of the supposed LC–MS/MS methodology

HPLC

Agilent 1200

Mass spectrometer

Agilent 6410 QQQ

Gradient mobile phase

A: H₂O (10 mM Ammonium formate,

pH:4.1)

Ionization source

Positive ESI

B: ACN

Drying gas: N₂ gas

Flow rate (12 L/min)

Pressure (55 psi)

Flow rate: 0.2 mL/min

Run time: 45 min

Injection volume: 20 µL

Agilent eclipse plus C₁₈ column

Length

50 mm

ESI temperature: 350 °C

Internal diameter

2.1 mm

Capillary voltage: 4000 V

Particle size

1.8 μm

Collision gas

High purity N₂

Temperature:

24 °C

Modes

Mass scan and product ion (PI)

Gradient system

Time

%B

Analyte

MST and its related in vivo phase I and phase II metabolites

0

5

40

40

Mass parameters

Fragmentor voltage: 130 V

43

40

45

5

Post time (15 min)

5

Collision energy of 20 eV