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Quantification of Acetylcholine in Cell Culture Systems by Semi-micro High-Performance Liquid Chromatography and Electrospray Ionization Mass Spectrometry

✍ Scribed by Acevedo, Larisa D.; Xu, Yaodong; Zhang, Xin; Pearce, Rosalyn J.; Yergey, Alfred

Publisher
John Wiley and Sons
Year
1996
Tongue
English
Weight
368 KB
Volume
31
Category
Article
ISSN
1076-5174

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✦ Synopsis

Electrospray ionization (ESI) was used to adapt the thermospray ionization liquid chromatographic/mass spectrometric (LC/MS) method of Liberato et al. to the determination of acetylcholine and choline released from cells in culture. A clonal cell line derived from a rat pheochromocytoma (PC12) was used and cultures depolarized in the presence of cholinesterase inhibitor to maximize the recovery of acetylcholine from the culture medium and protect against its hydrolysis. Concentration of acetylcholine and choline released into the incubation medium was accomplished with SepPak C,, cartridges. The preformed analyte cations were resolved by reversed-phase highperformance liquid chromatography and analyzed without derivatization by electrospray ionization mass spectrometry with the use of acetyll 'H,] choline as internal standard. LC/MS allows for the direct determination of acetylcholine and choline, which is not possible with other methods.

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