Metabolomics Portal

Parameter	Agilent 1290 Infinity I/II LC
Analytical columns	2X Agilent Zorbax Eclipse Plus C18, 1.8 µm, 100 x 2.1 mm column, p/n 959758-902
Inline filters	2X Agilent 1290 Infinity II inline filter, 0.3µm, p/n 5067-6189
Column temperature	45 °C
Injection volume	1 µL (equivalent to 0.1 µL plasma)
Autosampler temperature	20 °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 rates	0.4 mL/min
Gradient program	Pump A (G7120A)		Pump B (G7120A)
	Time	%B	Time	%B
	0	15	0	15
	2.5	50	1	15
	2.6	57	2	100
	9	70	6	100
	9.1	93	7	15
	11	96	12.9	15
	11.1	100
	12	100
	12.2	15
	12.9	15
Stop time	13 min
Post time	None
MCT Valve Position	Use current, Increase valve position after run

Chromatographic conditions for dual column setup