Metabolomics Portal

Parameter	Agilent 1290 Infinity I/II LC
Analytical column	Agilent Zorbax Eclipse Plus C18, 1.8 µm, 100 x 2.1 mm column, p/n 959758-902
Inline filter	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 rate	0.4 mL/min
Gradient program	Time	%B
	0	15
	2.5	50
	2.6	57
	9	70
	9.1	93
	11	96
	11.1	100
	12	100
	12.2	15
	16	15
Stop time	16 min
Post time	None

Chromatographic conditions for single column setup