Measurement of Acetylcholine Turnover Rate in Brain: An Adjunct to a Simple HPLC Method for Choline and Acetylcholine

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Abstract
An existing method for measuring acetylcholine (ACh) and choline (Ch) is shown to be useful formeasuring the turnover rate of ACh in mouse brain. Methl‐[3H]Ch is injected into mice. They are killed atdifferent times by microwave irradiation and Ch and AChextracted and separated by reverse‐phase HPLC. Ch andACh are converted to hydrogen peroxide by a post‐column enzyme reaction. Hydrogen peroxide, which isdirectly related to the tissue content of Ch or ACh, isdetermined electrochemically. The fractions that corre‐spond to the detector response for Ch and ACh are col‐lected for the measurement of radioactivity. In this wayspecific radioactivities of endogenous Ch and ACh areestimated in the same sample. We used the specific ra‐dioactivity values determined by this procedure to esti‐mate the turnover of ACh for striatum, cerebral cortex, and hippocampus of the mouse.