Table 1.

Optimized UHPLC and IM-qTOF parameters

Ultra-performance liquid chromatography (Agilent 1290 infinity II)
Column type	Poroshell 120 HILIC-Z (PEEK-coated)		SeQuant ZIC-pHILIC (PEEK-coated)
Parameters	2.1 mm × 50 mm, 2.7 μm		4.6 mm × 150 mm, 5 µm
Mobile phases	Positive ion mode	Negative ion mode	Positive ion mode	Negative ion mode
A	10 mM ammonium formate in water with 0.1% formic acid, pH 3	10 mM ammonium acetate in water, pH 9	10 mM ammonium formate in water with 0.1% formic acid, pH 3	10 mM ammonium acetate in water, pH 9
B	10 mM ammonium formate in water/ACN (1:9) with 0.1% formic acid, pH 3	10 mM ammonium acetate in water/ACN (1:9), pH 9	10 mM ammonium formate in water/ACN (1:9) with 0.1% formic acid, pH 3	10 mM ammonium acetate in water/ACN (1:9), pH 9
Gradient	Time (min)	% B	Time (min)	% B
	0.00	93	0	80
	1.80	80	15	20
	2.00	70	16	5
	2.30	70	20	5
	2.35	93	21	80
	3.50	93	30	80
Flow rate	0.8 mL/min		0.3 mL/min
Column temp	30 °C		30 °C
Injection vol	1 μL		5 μL

IM-qTOF (Agilent 6560)
Parameter		Parameter	Positive ion mode (V)	Negative ion mode (V)
Mass range	50–1700 m/z	Drift tube entrance volt.	1250	− 1175
IM transient rate	16 (IM transients/frame)	High pressure funnel RF	120	− 100
Max drift time	60 ms	Trap funnel RF	180	− 80
Trap fill time	2500 µs	Rear funnel RF	130	− 90
Trap release time	500 µs	Trap exit grid 1	88.5	− 86
Pulsing sequence length	4-bit	Trap exit grid 2	87	− 84.9

See Table S1 for a comprehensive list of IM-qTOF parameters