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HPLC Application Sheet
Choline & Acetylcholine by LC-MS

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A sensitive and specific method for the determination of Acetylcholine and its main decomposition product, Choline by LC-MS.

Method Conditions


Column Cogent UDC Cholesterol™, 4mm, 100A
Catalog No. 69069-7.5P
Dimensions 4.6 x 75 mm
Mobile Phase  90:10 acetonitrile/DI water + 0.5% foric acid (pH 3.48)
Flow rate 0.5 mL/minute
Injection Volume 1 µL
Samples Acetylcholine (m/z 146)
Choline (m/z 104)
(5 mg/mL (free base) of each dissolved in the mobile phase)
Detection Mass Spec: Atmospheric Pressure Chemical Ionization in positive mode: APCI+ Single Ion Monitoring


Discussion

These challenging quaternary amine compounds are well retained using a simple acetonitrile/DI water mobile phase with a low percentage of formic acid. A high level of the organic solvent helps to eliminate the problems associated with low aqueous solubility of the samples as well improves the sensitivity of MS detection. Presented separation can be used for analysis of low level quaternary amine compounds in biological and environmental samples. The impact of biological matrices needs to be evaluated.
Note: No ion paring reagent required in the above isocratic method. For reproducible results please use pre-mixed mobile phase and control the temperature of the column.





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