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LC-MS Isocratic Separation of Neurotransmitters on XBridge HILIC

LC-MS Isocratic Separation of Neurotransmitters on XBridge HILIC
  • Waters Corporation
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Two neuron and synapse, sending electrical and chemical signals

This is an Application Brief and does not contain a detailed Experimental section.

Abstract

This application brief demonstrates the LC-MS isocratic separation of neurotransmitters on XBridge HILIC Column.

Introduction

The compounds used in this study are:

  1. Acetylcholine (Ach)
  2. Choline (Ch)

Experimental

LC Conditions

Column:

XBridge HILIC, 2.1 x 50 mm, 3.5 μm

Part number:

186004432

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 diluent:

75:25 ACN:MeOH with 0.2% HCOOH

Sample concentration:

5 ng/mL each

Column temperature:

30 °C

Weak and strong needle wash:

95:5 ACN:H2O

Detection:

MS

Sampling rate:

5 points/sec

Instrument:

Waters ACQUITY UPLC with TQD

MS Conditions

Ionization mode:

ES+

Capillary:

0.5 kV

Cone:

40 V (choline), 25 V (acetylcholine)

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

Results and Discussion

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WA64076, August 2009

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