Table 2.

Ultra-high performance liquid chromatography gradient parameters for literature methods.

A)				B)				C)
HS30-LFRa				HS22-HFRb				HS25-LFRc
Time (min)	Flow (¼L/min)d	Solvent A (%)e	Solvent B (%)f	Time (min)	Flow (¼L/min)d	Solvent A (%)g	Solvent B (%)h	Time (min)	Flow (¼L/min)d	Solvent A (%)g	Solvent B (%)h
Initial	50	68	32	Initial	100	99	1	Initial	50	99	1
1.5		55	45	2		90	10	2		90	10
4		48	52	6		70	30	6		70	30
5		42	58	8		50	50	8		50	50
8		34	66	12		25	75	12		25	75
11		30	70	15		1	99	15		1	99
14		25	75	20		0	100	20		0	100
18		3	97	21		0	100	21		0	100
21		3	97	21.50		99	1	21.50		99	1
25		3	97	22		99	1	25		99	1
25.10		68	32
30		68	32

^aMethod adapted from Klatt et al. (2018).

^bMethod adapted from Raghunandan et al. (2019).

^cMethod modified from Raghunandan et al. (2019).

^dFlow rate of solvents (mobile phase).

^eSolvent composition: Acetonitrile/water (60/40, v/v) mixture containing ammonium formate (10 mM, pH 6.2) and formic acid (0.1%).

^fSolvent composition: 2-propanol/acetonitrile (90/10, v/v) mixture containing ammonium formate (10 mM, pH 6.2) and formic acid (0.1%).

^gSolvent composition: 100% water containing 0.1% formic acid.

^hSolvent composition: 100% acetonitrile containing 0.1% formic acid.