C4D Analysis of Acetylchorine Chloride on a Prince CE System
The eDAQ C4D system was connected to the PrinCE-C 600 capillary electrophoresis instrument and acetylcholine chloride was analysed.
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.
- Analyte: Acetylcholine chloride (AChCl) dissolved in water, analysed at the concentrations 10 mM, 50 mM, 100 mM, 500 mM and 1000 mM
- MES/Arg/CTAB with pH 5.7, where MES is 2-(N-morpholino)ethanesulfonic acid, Arg is L(+)-Arginine, and CTAB is hexadecyltrimethylammonium bromide
- separation voltage = 30 kV
- recorded separation current = 11 – 12 µA
- 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
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.