LC-MS/MS chromatographic parameters.

LC			MS/MS
LC	Agilent RRLC 1200		MS	Agilent 6410 Triple Quadrupole
Gradient mobile phase	A: 5 mM ammonium formate in H2O		ESI	Positive ESI
	B: acetonitrile			Drying nitrogen gas
	0.3 mL min−1			11 L min−1 flow rate
	65 min			55 psi
C18 column (Agilent Eclipse Plus)	150 mm length			350 °C
	2.1 mm ID			4000 V capillary voltage
	3.5 μm particle size		Modes	MS scan and PI
	23 ± 2 °C		Collision gas	High-purity nitrogen gas
Gradient mobile phase	Time	% Acetonitrile	Drug	X-376 and its related metabolites
	0	5
	5	5	Mass analyzer	30 eV collision energy
	40	60		200 ms dwell time
	60	90		500 ms scan time
	65	5		145 V fragmentor voltage