LC-MS Isocratic Separation of Neurotransmitters on Atlantis HILIC Silica

Column:

Atlantis HILIC Silica, 2.1 x 50 mm, 3 μm

Part Number:

186002011

Mobile Phase A:

10 mM NH₄COOH with 0.125% HCOOH in H₂O

Mobile Phase B:

10 mM NH₄COOH with 0.125% HCOOH in 90:5:5 ACN:MeOH:H₂O

Isocratic Mobile Phase Composition:

10% A; 90% B

Flow Rate:

0.5 mL/min

Injection Volume:

10.0 μL (full loop)

Sample Concentration:

5 ng/mL each

Sample Diluent:

75:25 ACN:MeOH with 0.2% HCOOH

Column Temp.:

30 °C

Weak and Strong Needle Wash:

95;5 ACN:H₂O

Detection:

MS

Sampling Rate:

5 points/sec

Instrument:

Waters ACQUITY UPLC with TQD

Ionization Mode:

ES+

Capillary:

0.5 kV

Cone:

25 V (acetylcholine), 40 V (choline)

Source Temperature:

120 °C

Desolvation Temperature:

350 °C

Desolvation Gas Flow:

800 L/Hr

SIR:

146.1 m/z (acetylcholine); 104.0 m/z (choline)

Dwell Time:

150 msec

ISD:

10 msec

ICD:

10 msec