HPLC ELSD Method for Analysis Acetylcholine and Choline on Obelisc R Column

HPLC Method for Analysis of Acetylcholine, Choline on Obelisc R by SIELC Technologies

HPLC Method for Analysis of Acetylcholine, Choline on Obelisc R Column by SIELC Technologies

 High Performance Liquid Chromatography (HPLC) Method for Analysis of Acetylcholine, Choline

Acetylcholine and choline are related compounds that play distinct roles in the body, particularly in the nervous system.

  1. Choline:
  • Definition: Choline is an essential nutrient that belongs to the B-vitamin complex.
  • Chemical Structure: Choline is a quaternary ammonium salt containing a positively charged nitrogen atom and is often written as (N(CH_3)_3^+).
  • Function: Choline serves as a precursor for the synthesis of various important molecules, including phospholipids (essential components of cell membranes) and the neurotransmitter acetylcholine. Choline is crucial for maintaining cell membrane integrity and is involved in liver function.
  • Dietary Sources: Choline is found in foods such as eggs, meat, fish, nuts, and certain vegetables. It can also be consumed as a dietary supplement.
  1. Acetylcholine:
  • Definition: Acetylcholine is a neurotransmitter, which is a chemical messenger that transmits signals between nerve cells (neurons) or from neurons to muscles.
  • Chemical Structure: Acetylcholine is derived from the combination of choline and acetyl coenzyme A. Its chemical formula is (CH_3COOCH_2CH_2N(CH_3)_3^+).
  • Function: Acetylcholine plays a crucial role in transmitting signals across synapses, including those between neurons and muscles (neuromuscular junctions). It is involved in muscle contraction, autonomic nervous system functions, and cognitive processes such as memory and learning.
  • Synthesis and Release: Acetylcholine is synthesized within nerve cells and released into synapses upon a nerve impulse. Its action is terminated by the enzyme acetylcholinesterase, which breaks it down in the synaptic cleft.

In summary, choline is a nutrient that the body uses to synthesize acetylcholine, among other important molecules. Acetylcholine, on the other hand, is a neurotransmitter responsible for transmitting signals in the nervous system. While choline is obtained from the diet or supplements, acetylcholine is a key mediator of nerve and muscle function in the body.

Acetylcholine and Choline can be retained, separated and analyzed using an Obelisc R mixed-mode stationary phase column. The analysis employs an isocratic method with a simple mobile phase consisting of water, acetonitrile (MeCN), and ammonium acetate as a buffer. Detection is achieved using ELSD

ColumnObelisc R, 2.1 x 100 mm, 5 µm, 100 A
Mobile PhaseMeCN/H2O – 60/40%
BufferAmmonium acetate pH 5.0 – 10 mM
Flow Rate0.2 ml/min
DetectionELSD, the nebulizer and evaporator temperatures 70°C,
 with a gas flow rate of 1.6 Standard Liters per Minute (SLM)
Samples0.5 mg/ml
Injection volume1 µl
LOD*200 ppb
* LOD was determined for this combination of instrument, method, and analyte, and it can vary from one laboratory to another even when the same general type of analysis is being performed.

Class of Compounds
Quaternity amines
Analyzing CompoundsAcetylcholine, Choline

Application Column

Obelisc R

Column Diameter: 2.1 mm
Column Length: 100 mm
Particle Size: 5 µm
Pore Size: 100 A

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Application Analytes:

Application Detection:
ELSD Detection
SIELC Technologies usually develops more than one method for each compound. Therefore, this particular method may not be the best available method from our portfolio for your specific application. Before you decide to implement this method in your research, please send us an email to research@sielc.com so we can ensure you get optimal results for your compound/s of interest.