C4D Analysis of Acetylchorine Chloride on a Prince CE System

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Figure 1. AChCl peaks at different concentrations
Figure 2. Calibration curve

The eDAQ C4D system was connected to the PrinCE-C 600 capillary electrophoresis instrument and acetylcholine chloride was analysed.

Introduction

The eDAQ C4D system was connected to the PrinCE-C 600 capillary electrophoresis (CE) instrument as described in the application note C4D004. Acetylcholine chloride (AChCl) was then analysed and a calibration curve was produced. The CE instrument was set to negative mode and the C4D system measured the Cl- anions.

Conditions

  • Analyte: Acetylcholine chloride (AChCl) dissolved in water, analysed at the concentrations 10 mM, 50 mM, 100 mM, 500 mM and 1000 mM
  • Buffer:
    • MES/Arg/CTAB with pH 5.7, where MES is 2-(N-morpholino)ethanesulfonic acid, Arg is L(+)-Arginine, and CTAB is hexadecyltrimethylammonium bromide
  • Instrument:
    • separation voltage = 30 kV
    • recorded separation current = 11 – 12 µA
  • Capillary:
    • outer diameter = 360 µm
    • internal diameter = 25 µm
    • length = 56 cm
    • length to detector = 42 cm
  • Injection: pressure 300 mbar for 10 seconds
  • C4D settings
    • frequency = 500 kHz
    • amplitude = 100 A
    • headstage gain OFF
  • Data recording:
    • lowpass filter = 1 Hz
    • range = 50 mV
    • sampling rate = 100 data points per second

Data Analysis

Figure 1 shows the AChCl peaks at the different concentrations. Figure 2 shows how the software can automatically produce a calibration curve by plotting the peak areas versus known concentrations of analyte.