HPLC ELSD Method for Analysis of Acetylcholine on Obelisc R Column

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

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

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

Acetylcholine is a neurotransmitter, which is a chemical messenger that transmits signals across synapses, the gaps between nerve cells or between nerve cells and muscles. It plays a crucial role in the nervous system, both in the central nervous system (CNS) and the peripheral nervous system (PNS).

Here are key points about acetylcholine:

  1. 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^+).
  2. Synthesis and Release: Acetylcholine is synthesized within nerve cells through a series of enzymatic reactions. Upon a nerve impulse reaching the end of a nerve cell, acetylcholine is released into the synapse.
  3. Neuromuscular Junction: In the peripheral nervous system, acetylcholine is the neurotransmitter responsible for transmitting signals from motor neurons to muscle fibers at the neuromuscular junction. The binding of acetylcholine to receptors on the muscle cell membrane triggers muscle contraction.
  4. Autonomic Nervous System: In the autonomic nervous system, acetylcholine is involved in transmitting signals in the parasympathetic branch. It slows the heart rate, stimulates digestive processes, and promotes a “rest and digest” response.
  5. Cognitive Functions: In the central nervous system, acetylcholine is implicated in various cognitive functions, including memory, learning, and attention. Its role in the brain is complex and involves interactions with different receptor types.
  6. Degradation: The enzyme acetylcholinesterase breaks down acetylcholine in the synaptic cleft, terminating its action and preventing continuous stimulation.
  7. Medicinal Importance: Drugs that affect acetylcholine receptors or inhibit acetylcholinesterase activity are used for various medical purposes. For example, some medications aim to enhance acetylcholine activity in Alzheimer’s disease.

The balance of acetylcholine and other neurotransmitters is essential for proper nervous system function. Imbalances in acetylcholine levels have been implicated in various neurological disorders.

Acetylcholine can be retained 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 – 10/90%
BufferAmmonium acetate pH 5.0 – 10 mM
Flow Rate0.2 ml/min
DetectionELSD, the nebulizer and evaporator temperatures 50°C,
 with a gas flow rate of 1.6 Standard Liters per Minute (SLM)
Samples5 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

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.