RPLC-MS Method

1. RPLC-MS Method for MS1 only

Column parameter

Waters, UPLC HSS T3 column (1.8 μm, 2.1× 100 mm)

Mobile phase

Positive mode

A:100% H2O (0.1% Formic acid)

B:100% ACN (0.1% Formic acid)

Negative mode

A:100% H2O (0.5 mM NH4F)

B:100% ACN

Gradient parameter

Time Flow rate (uL/min) A % B %
0.00 500 99.0 1.0
1.00 500 99.0 1.0
8.00 500 1.00 99.0
10.00 500 1.00 99.0
10.00 500 99.0 1.0
12.00 500 99.0 1.0

2. DDA

Column parameter

Waters, UPLC HSS T3 column (1.8 μm, 2.1× 100 mm)

Mobile phase

Positive mode

A:100% H2O (0.1% Formic acid)

B:100% ACN (0.1% Formic acid)

Negative mode

A:100% H2O (0.5 mM NH4F)

B:100% ACN

Gradient parameter

Time Flow rate (uL/min) A % B %
0.00 300 99.0 1.0
1.50 300 99.0 1.0
13.00 300 1.00 99.0
16.50 300 1.00 99.0
16.60 300 99.0 1.0
20.00 300 99.0 1.0