Acetylcholine measurement by high-performance liquid chromatography using an enzyme-loaded postcolumn reactor

Area of Science:

• Biochemistry
• Analytical Chemistry
• Enzyme Technology

Background:

• Acetylcholine is a crucial neurotransmitter.
• Accurate measurement of acetylcholine is vital for neurological research.
• Enzyme-based detection methods offer high sensitivity.

Purpose of the Study:

• To develop a stable and sensitive method for acetylcholine measurement.
• To evaluate the long-term stability of choline oxidase and cholinesterase immobilized on anion-exchange cartridges.

Main Methods:

• Immobilization of choline oxidase and cholinesterase onto anion-exchange cartridges.
• High-performance liquid chromatography (HPLC) separation of acetylcholine.
• Postcolumn enzymatic conversion of acetylcholine to hydrogen peroxide (H2O2).
• Electrochemical detection of H2O2.

Main Results:

• Immobilized enzymes retained activity for 1-2 weeks at room temperature.
• The method allowed for sensitive acetylcholine detection in tissue extracts.
• HPLC separation at pH 7 was effective for sample preparation.

Conclusions:

• Enzyme-adsorbed anion-exchange cartridges provide a stable platform for acetylcholine analysis.
• This method offers a robust approach for quantifying acetylcholine in biological samples.
• The developed technique enhances capabilities in neurotransmitter research.