Table 1 Experimental settings of mass spectrometry analyses

LC settings:

Separation column

MonoCap

75 μm × 150 mm,

C18, monolith

Michrom

75 μm × 50 mm, C18, 3 μm

Zorbax

75 μm × 150 mm,

C18, 5 μm

MonoCap

50 μm × 250 mm,

C18, monolith

KYA

100 μm × 150 mm,

C18, 5 μm

Flow rate (nL/min)

200

500

300

200

300

Mobile phase gradient

2%B (0 min)

→ 40%B (100 min)

→ 100%B (100.1 min)

→ 100%B (120 min)

→ 2%B (120.1 min)

→ 2%B (140 min)

5%B (0 min)

→ 45%B (80 min)

→ 95%B (81 min)

→ 95%B (89 min)

→ 5%B (90 min)

→ 5%B (120 min)

3%B (0 min)

→ 40%B (50 min)

→ 100%B (55 min)

→ 100%B (65 min)

→ 3%B (66 min)

→ 3%B (80 min)

0%B (0 min)

→ 50%B (140 min)

→ 50%B (155 min)

→ 100%B (156 min)

→ 100%B (170 min)

0%B (0 min)

→ 50%B (60 min)

→ 100%B (80 min)

→ 100%B (90 min)

Data acquisition time

(min)

120

90

65

170

90

MS settings:

Instrument

Hitachi NanoFrontier LD

Thermo LTQ-Orbitrap Discovery

Agilent 6510 Q-TOF

GE& Thermo MDLC-LTQ

ABI 4800 MALDI-TOF/TOF

MS scan

TOF

Orbitrap

TOF

LIT

TOF

MS/MS scan

TOF

LTQ

TOF

LIT

TOF

MS/MS spectrum acquisition

IBA mode _b)

IBA mode

IBA mode

IBA mode

IBA mode