Metabolomics Portal - Methods Brain Lipidomics

Parameter	Agilent 1290 Infinity II LC
Analytical column	Agilent Zorbax Eclipse Plus C18, 1.8 µm, 100 x 2.1 mm column, p/n 959758-902
Inline filter	InfinityLab Quick Change Inline Filters, 0.2µm, p/n 5067-1603
Column temperature	45 °C
Injection volume	1-5 µL (sample dependent)
Autosampler temperature	17 °C
Needle wash	Wash vial, 3 s, 1:1 butanol/methanol
Needle wash	*Alternatively, flush port, 10 s, 1:1 mobile phase A/B can be used
Mobile phase	A = 10 mM ammonium formate, 5µM Agilent Deactivator Additive (5191-3940) in 5:3:2 water/acetonitrile/2-propanol
Mobile phase	B = 10 mM ammonium formate in 1:9:90 water/acetonitrile/2-propanol
Flow rate	0.4 mL/min
Gradient program	Time	%B
	0	15
	2.5	50
	2.6	57
	12	70
	12.5	80
	15.4	90
	15.5	100
	16.5	100
	17	15
	20	15
Stop time	20 min
Post time	None
Gradient program	0-1 minutes	To waste
	1-18.5 minutes	To MS
	18.5-20 minutes	To waste

Chromatographic conditions