UPLC-MS Analysis of Mono-, Di- and Oligosaccharides Using ACQUITY UPLC BEH Amide Columns

Column:

ACQUITY UPLC BEH Amide 2.1 x 50 mm, 1.7 μm

Part number:

186004800

Mobile phase A:

80/20 MeCN/H₂O with 0.10% ammonium hydroxide [NH₄OH]

Mobile phase B:

30/70 MeCN/H₂O with 0.10% ammonium hydroxide [NH₄OH]

Flow rate:

0.17 mL/min

Gradient:

5 minute gradient, 80%–50% MeCN with 10-minute re-equilibration

Injection volume:

0.7 μL (PLNO)

Sample concentration:

10 μg/mL each

Sample diluent:

50/50 MeCN/H₂O

Column temperature:

35 °C

Strong needle wash:

20/80 MeCN/H₂O (800 μL)

Weak needle wash:

75/25 MeCN/H₂O (500 μL)

Seal wash:

50/50 MeCN/H₂O

Instrument:

Waters ACQUITY UPLC with ACQUITY TQD

Ionization mode:

ES

Capillary:

2.8 kV

Cone voltage:

25 V

Source temperature:

120 °C

Desolvation temperature:

350 °C

Desolvation gas flow:

500 L/Hr

Cone:

50 L/Hr

SIR (m/z):

179.2 (fructose, glucose);

341.3 (sucrose, maltose);

503.4, 665.5, 827.6, 989.7, 1151.8 (maltooligosaccharides [n=1 to 5])

Dwell time:

0.08 s