UPLC-MS Analysis of Stevia Related Compounds Using ACQUITY UPLC BEH Amide Columns

Column:

ACQUITY UPLC BEH Amide 2.1 x 100 mm, 1.7 µm

Part Number:

186004801

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:

0.20 mL/min

Flow Profile:

95% A/5% B (77.5% MeCN with 0.10% NH₄OH)

Injection Volume:

1.3 μL (PLNO)

Sample Concentration:

50 μg/mL stevia, 10 μg/mL truvia

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:

ESCapillary: 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)

317.5 (steviol);

803.8 (stevioside);

950.1 (rebaudioside C);

966.1 (rebaudioside A)

Dwell Time:

0.08 s